canavalia gladiata - shodhgangashodhganga.inflibnet.ac.in/bitstream/10603/6744/10/10_chapter...

Canavalia gladiata - Results

Institute of Pharmaceutical Technology, SPMVV, Tirupati. 76

CANAVALIA GLADIATA

Results of Pharmacological Studies:

Acute toxicity and gross behavioral changes:

Alcoholic extract and its fractions of Canavalia gladia found to be safe since no

animal died even at the maximum single dose of 3200mg/kg, orally. The animals did

not show any significant gross behavioral changes at the doses tested. Hence, doses of

the alcoholic extract and its fractions selected for the present study were 100, 200,

300mg/kg, orally.

Assessment of Neuropharmacological Studies:

Alcoholic extract of Canavalia gladiata (CG) at a dose of 100, 200, 300 mg/kg

showed significant (P< 0.01) and dose dependent increase in spontaneous motor

activity as compared to control vehicle group (Fig. 2).

Alcoholic extract at given doses showed no significant change in reaction time

as compared to control group in Eddys hot plate (Fig. 3).

The animals treated with 100, 200, 300 mg/kg of CG exhibited significant

(P<0.01) dose dependent increased motor co-ordination (Fig.4).

The results of forced swimming test (FST) showed that there was significant

decrease (P<0.01) in immobility time and significant (P<0.01) increase of swimming

time of animals at 100, 200, 300 mg/kg as compared to control vehicle group (Fig.5

&6).

At given doses alcoholic extract exhibited significant (P<0.01) dose dependent

decreased immobility time as compared to vehicle group in tail suspension test (TST)

(Fig.7).



The results of the hole board test revealed that there was a significant (P<0.01)

increase in number of head dipping as compared to control vehicle group (Fig.8).

Antiparkinsonian Activity:

Behavioural Study of Mice after MPTP treatment:

Spontaneous motor activity was significantly (0.001) decreased in MPTP

treated group as compared to control group. Alcoholic extract at given doses

significantly increased the locomotor activity but maximum activity reached on 6th

day of treatment as compared to MPTP treated animals (Table.2; Fig.9).

Retention time on rotarod was significantly decreased in the MPTP treated

group as compared with the control group and it was significantly (P<0.01) dose

dependently improved on 1st

, 3rd

and 6th

day with alcoholic extract of CG treatment

(Table.2; Fig.10).

The results of the hole board test revealed that there was a significant decrease

in the number of head dippings in MPTP treated mice. It was significantly (0.01)

reversed with alcoholic extract of CG as compared to MPTP treated group (Table.2;

Fig.11).

Effect on brain dopamine level:

The results of the present study showed that the dopamine level was

significantly (P<0.001) decreased in the MPTP treated animals at 20 mg/kg, i.p in

compared to control vehicle group while their levels significantly (P<0.001) dose

dependent increased in alcoholic extracts of canavalia gladia (AECG) at a dose of 200

and 300 mg/kg as compared to MPTP treated groups (Table.3; Fig.12).



On treatment with Pet. ether extract of Canavalia gladiata (PEECG) exhibited

significant (P<0.01, 0.001) increase of dopamine level at 200 and 300 mg/kg when

compared to MPTP treated group (Table.3; Fig.13).

Ethyl acetate extract of Canavalia gladiata (EAECG) showed significant

(P<0.001) increase in dopamine level at 200 and 300 mg/kg as compared to lower

doses (Table.3; Fig.14).

Water (aqueous) extract of Canavalia gladiata exhibited significant (P<0.001)

dose dependent increase of dopamine level at given doses as compared to MPTP

treated group (Table.3; Fig.15).

Effect on other brain amines:

Epinephrine (E):

Epinephrine level was significantly (P<0.001) reduced in MPTP treated animals

compared to control vehicle group. Epinephrine level is restored significantly

(P<0.001) dose dependently with alcoholic extract at given doses as compared to

MPTP treated group (Table.4; Fig.16).

Animals treated with PEECG exhibited a significantly (P<0.01, 0.001)

increased level of epinephrine at 100 mg, 200 and 300 mg as compared to MPTP


EAECG showed significant (P<0.001) dose dependent increase in the level of

epinephrine as compared to MPTP neurotoxicity group (Table.4; Fig.18).

Epinephrine level was significantly (P<0.001) elevated with aqueous extract of

CG at given doses (Table.4; Fig.19).



Norepinephrine (NE):

The result of the present study showed that NE level was significantly

(P<0.001) reduced in MPTP treated mice as compared to vehicle treated group. While

their levels significantly (P<0.001) dose dependently protected with alcoholic extract as


PEECG exhibited significant (P<0.001) increase in the level of NE only at a

dose of 300mg/kg (Table.5; Fig.21).

NE level was increased significantly (P<0.001) with ethyl acetate extract at 200

and 300 mg/kg as compared to MPTP treated group (Table.5; Fig.22).

The animals treated with aqueous extract showed markedly significant (P<0.05;

P<0.001) increase of NE with lower and higher doses as compared to MPTP group

(Table.5; Fig.23).

Serotonin:

Results of the Present study showed significant (P<0.001) reduction of

serotonin level in MPTP treated animals as compared to control group. Subsequently

their level was significantly (P<0.001) dose dependently enhanced with alcoholic

extract of CG (Table.6; Fig.24).

The animals treated with PEECG produced significant (P<0.001) improvement

of serotonin at a dose of 300 mg/kg while no significant change at dose 100 and 200

mg/kg as compared to MPTP group (Table.6; Fig.25).

Serotonin level was elevated significantly (P<0.05, 0.01, 0.001) with EAECG at

a dose of 100, 200, 300 mg/kg as compared to MPTP group (Table.6; Fig.26).



Treatment with aqueous extract of CG exhibited significant (P<0.001) increase

in serotonin level at a dose of 200 and 300 mg/kg as compared to MPTP group

(Table.6; Fig.27).

Results of Invivo antioxidant Studies:

Effect on Lipid Peroxidation:

Results of the present study showed that brain MDA level was significantly

(P<0.001) elevated in the MPTP treated group as compared to the control vehicle

group. Alcoholic extract of CG at a dose of 200 and 300 mg/kg showed significant

(P<0.001) dose dependent decrease of MDA level as compared to lower dose and

MPTP treated group (Table.7; Fig.28).

Mice treated with PEECG exhibited significant (P<0.001) reduction of MDA

level at 300 mg/kg as compared to MPTP treated group (Table.7; Fig.29).

Animals treated with EAECG showed significant (P<0.05, 0.001) reduction of

MDA level at a dose of 200 and 300 mg/kg compared to MPTP treated group (Table.7;

Fig.30).

Treatment with aqueous extract of CG showed significant (P<0.01, 0.001)

decrease at a dose of 200 and 300 mg/kg as compared to MPTP treated group (Table.7;

Fig31).

Effect on Reduced glutathione level:

Results of the present study showed, reduced glutathione level was significantly

(P<0.001) decreased in MPTP treated group as compared to the control vehicle group.

GSH level was restored significantly (P<0.001) dose dependently at 200 and 300 mg/kg

with AECG as compared to MPTP treated group (Table.8; Fig.32).



Animals treated with the PEECG showed moderate activity (P<0.01) at a dose

of 300 mg/kg as compared to MPTP treated group (Table.8; Fig.33).

Treatment with EAECG exhibited significant (P<0.001) increase the level of

reduced glutathione at 300 mg/kg as compared to MPTP treated animals (Table.8;

Fig.34).

At given doses aqueous extract exhibited significant (P<0.001) increase of

reduced glutathione level as compared to MPTP treated group (Table.8; Fig.35).



Table. 2: Effect of Alcoholic extracts of Canavalia gladiata seed on Spontaneous motor activity, Grip strength, Alertness (Hole

Board test) in MPTP treated mice.

Groups

Spontaneous motor activity score

Grip Strength in Seconds

Alertness (no. of head dippings)

1st day 3

rd day 6

th day

1st day 3

rd day 6

th day 1

st day 3

rd day 6

th day

Control 446.6 ± 12.0 446.6 ± 12.0 446.6 ± 12.0 120 ± 0 120 ± 0 120 ± 0 50.5 ± 2.2 50.5 ± 2.2 50.5 ± 2.2

MPTP 278.5 ± 7.0

***

179.8 ± 5.3

***, +++

90.3 ± 6.2

***, +++ 83.3 ± 2.8

***

33.6 ± 7.2

***

13.8 ± 1.1

***

22.3 ± 1.60

***

14.0 ± 2.1

***

8.6 ± 1.2

***

100mg/kg CGE

+ MPTP

298.5 ± 1.8

***

291.7 ± 3.3

***, +++

296.1 ± 2.1

***, +++

90.1 ± 2.4

***

91.6 ± 1.5

***, +++

97.8 ± 0.9 ***, +++

30.1 ± 1.1

***, ++

33.8 ± 1.4

***, +++

38.0 0.7

***, +++

200 mg/kg CGE

+ MPTP

305.1 ± 3.0

*** , +

3.7.8 ± 0.9

***, +++

318.5 ± 1.7

***, +++

99.6 ± 1.3

***, +++

104.3 ± 2.5

*, +++

106.6 ± 3.3 ***, +++

37.7 ± 0.9

***,+++

41.6 ± 1.6

*, +++

43.1 ± 1.7

*, +++

300 mg/kg CGE

+ MPTP

310.0 ± 1.5

***,++

329.8 ± 2.7

***, +++

334.3 ± 1.7

***, +++

102.5 ± 2.7 *** , +++

107.5 ± 1.4

+++

111.6 ± 1.3

*, +++

38.0 ± 1.0

***, +++

45.0 ± 1.4

+++

48.1 ± 1.4

+++

300 mg/kg CGE 334.6 ± 1.8

***, +++

340.6 ± 1.1

***, +++

349.5 ± 1.7

***, +++ 114.0 ± 1.9

+++

116.3 ± 1.0

+++

117.3 ± 0.8 ***, +++

45.3 ± 1.5

***, +++

49.0 ± 0.9

+++

53.5 ± 1.5

+++

Values are expressed as Mean ± SEM (n = 6); *** (P< 0.001) Vs Control group; ++ (P< 0.01), +++ (P< 0.001) Vs MPTP group.



Table.3: Effect of Canavalia gladiate seed on brain dopamine level in MPTP treated mice.

Groups

g/g brain tissue

Alcoholic Petroleum ether Ethyl acetate Water extract

Control (Saline) 3.815 ± 0.055 3.815 ± 0.055 3.815 ± 0.055 3.815 ± 0.055

MPTP 1.257 ± 0.038*** 1.257 ± 0.038*** 1.257 ± 0.038*** 1.257 ± 0.038***

100mg/kg CGSE + MPTP 1.506 ± 0.029*** 1.308 ± 0.037*** 1.395 ± 0.100*** 1.444 ± 0.032***

200 mg/kg CGSE + MPTP 2.046 ± 0.027***

+++

1.610 ± 0.051***

++

2.009± 0.031***

+++

2.031 ± 0.049***

+++

300 mg/kg CGSE + MPTP 3.093 ± 0.022***

+++

2.591 ± 0.099***

+++

3.036 ± 0.056***

+++

3.064 ± 0.034***

+++

300 mg/kg CGSE 3.121 ± 0.042*

+++

2.881 ± 0.064***

+++

3.028 ± 0.058***

+++

3.090 ± 0.039***

+++

Values are expressed as Mean ± SEM (n = 6);

* (P< 0.05), *** (P< 0.001) Vs Control group;

++ (P< 0.01), +++ (P< 0.001) Vs MPTP group.



Table.4: Effect of Canavalia gladiata seed on brain epinephrine level in MPTP treated mice.

Groups

g/g brain tissue


Control 3.288 ± 0.1 3.288 ± 0.10 3.288 ± 0.10 3.288 ± 0.10

MPTP 2.106 ± 0.09*** 2.106 ± 0.09*** 2.106 ± 0.09*** 2.106 ± 0.09 ***

100mg/kg CGE + MPTP 2.861 ± 0.06**

+++

2.563 ± 0.06***

++

2.749 ± 0.04 ***

+++

2.779 ± 0.04 **

+++

200 mg/kg CGE + MPTP 2.959 ± 0.03 *

+++

2.824 ± 0.06 **

+++

2.878 ± 0.05 *

+++

2.932 ± 0.09 *

+++

300 mg/kg CGE + MPTP 3.136 ± 0.06 +++ 2.849 ± 0.04 **

+++

2.992 ± 0.03 +++ 3.052 ± 0.05+++

300 mg/kg CGE 3.211 ± 0.06 +++ 3.023 ± 0.03+++ 3.072 ± 0.10+++ 3.178 ± 0.07 +++


*(P< 0.01), ** (P< 0.01), *** (P< 0.001) Vs Control group;

++ (P< 0.01), +++ (P< 0.001) Vs MPTP group.



Table.5: Effect of Canavalia gladiata seed on brain norepinephrine level in MPTP treated mice.

Groups

g/g brain tissue


Control 3.344 ± 0.054 3.344 ± 0.054 3.344 ± 0.054 3.344 ± 0.054

MPTP 2.148 ± 0.041*** 2.148 ± 0.041*** 2.148 ± 0.041*** 2.148 ± 0.041***

100mg/kg CGE + MPTP 2.589 ± 0.068***

+++

2.058 ± 0.065***

2.168 ± 0.078***

2.418 ± 0.057 ***

+

200 mg/kg CGE + MPTP 2.859 ± 0.031***

+++

2.274 ± 0.054***

+++

2.699 ± 0.049 ***

+++

2.827 ± 0.051***

+++

300 mg/kg CGE + MPTP 3.150 ± 0.066 +++ 2.602 ± 0.064**

+++

3.090 ± 0.068+++ 3.126 ± 0.051+++

300 mg/kg CGE 3.304 ± 0.028+++ 3.004 ± 0.029**

+++

3.104 ± 0.043+++ 3.188 ± 0.038 +++


** (P<0.01), *** (P< 0.001) Vs Control group;

+ (P<0.05), +++ (P< 0.001) Vs MPTP group.



Table.6: Effect of Canavalia gladiate seed on brain serotonin level in MPTP treated mice.

Groups

g/g brain tissue


Control 3.407 ± 0.050 3.407 ± 0.050 3.407 ± 0.050 3.407 ± 0.050

MPTP 2.068 ± 0.060*** 2.068 ± 0.060*** 2.068 ± 0.060*** 2.068 ± 0.060***

100mg/kg CGE + MPTP 2.458 ± 0.068***

+++

2.080 ± 0.050***

2.312 ± 0.052***

+

2.348 ± 0.052***

+

200 mg/kg CGE + MPTP 2.823 ± 0.030***

+++

2.141 ± 0.062***

2.412 ± 0.032***

++

2.621 ± 0.090***

+++

300 mg/kg CGE + MPTP 3.011 ± 0.036***

+++

2.476 ± 0.038***

+++

2.865 ± 0.050***

+++

2.980 ± 0.051***

+++

300 mg/kg CGE 3.201 ± 0.035 +++ 3.028 ± 0.075**

+++

3.085 ± 0.051**

+++

3.110 ± 0.049*

+++


* (P<0.05), ** (P<0.01), *** (P< 0.001) Vs Control group;

+ (P<0.05), ++ (P<0.01), +++ (P< 0.001) Vs MPTP group.



Table.7: Effect of Canavalia gladiata seed on lipid peroxidation (nmol/mg protein/hr) level in MPTP treated mice.

Groups

nmol/mg protein/hr


Control 26.25 ± 3.4 26.25 ± 3.4 26.25 ± 3.4 26.25 ± 3..4

MPTP 67.0 ± 5.6*** 67.0 ± 5.6*** 67.0 ± 5.6*** 67.0 ± 5.6***

100mg/kg CGE + MPTP 52.16 ± 4.3*** 60.750 ± 1.1*** 57.08 ± 3.0*** 56.45 ± 2.8***

200 mg/kg CGE + MPTP 37.000 ± 3.1 +++ 47.951 ± 2.8 **

++

44.87 ± 5.7*

+

43.50 ± 3.6*

++

300 mg/kg CGE + MPTP 29.833 ± 1.6 +++ 42.62 ± 3.7 *

+++

36.02 ±1.9 +++ 31.79 ± 1.3+++

300 mg/kg CGE 24.41 ± 2.3 +++ 31.87 ± 1.7 +++ 28.50 ± 1.9 +++ 27.58 ± 1.4 +++

Values are expressed as Mean ± SEM;


+ (P<0.05), +++ (P< 0.001) Vs MPTP group.



Table.8: Effect of Canavalia gladiate seed on reduced glutathione (µmol/mg protein) level in MPTP treated mice.

Groups

µmol/mg protein


Control 759.90 ± 53.2 759.90 ± 53.2 759.90 ± 53.2 759.90 ± 53.2

MPTP 476.60 ± 24.9*** 476.60 ± 24.9*** 476.60 ± 24.9*** 476.60 ± 24.9***

100mg/kg CGE + MPTP 599.27 ± 6.9**

+

494.08 ± 4.1***

504.82 ± 13.1 ***

555.70 ± 17.3 ***

+

200 mg/kg CGE + MPTP 655.60 ± 19.8 +++ 506.92 ± 8.4***

660.67 ± 11.3*** 614.02 ± 22.2**

+++

300 mg/kg CGE + MPTP 726.0 ± 4.9 +++ 620.37 ± 9.3**

++

660.67 ± 15.3+++ 685.53 ± 14.8 +++

300 mg/kg CGE 745.0 ± 9.7 +++ 712.63 ± 9.7 +++ 730.20 ± 4.7+++ 737.63 ± 2.8+++


** (P<0.01), *** (P< 0.001) Vs Control group;

+ (P< 0.05), ++ (P< 0.01), +++ (P< 0.001) Vs MPTP group.

Canavalia gladiata – Results


Results of Invitro antioxidant activity:

Free radical scavenging activity was carried out in different invitro models.

Several concentrations ranging from 3-800 µg/ml of the AECG seed were tested.

Results of present study showed that free radicals were scavenged by the test

compounds in a concentration dependent manner upto the given concentration in all the

models. The IC50 value of DPPH, nitric oxide, lipid peroxidation and superoxide

dismutage were found to be 300, 510, 320, 340 µg/ml respectively (Table.9; Fig. 36,

37, 38, 39).

Table.9 Effect of alcoholic extracts of Canavalia gladiata on different antioxidant

models

S.No.

% inhibition

Conc

µg/ml

DPPH Superoxide

dismutase

Nitric oxide Lipid

Peroxidation

1 3 14.69 7.3 11.99 10.85

2 6 18.9 9.8 16.0 13.3

3 12 22.4 13.5 21.8 21

4 25 26.7 17.4 24 24

5 50 35.4 19.8 34 26

6 100 38.9 22.2 39 43

7 200 40.2 30.2 54 52

8 400 75.4 39 57 59

9 800 89 68.5 86 90

10 IC50 (µg/ml) 300 510 320 340

(Values are mean of 3 replicates)



Result of Pharmacognostic Study:

Preliminary Phytochemical analysis:

Result of the present study showed the presence of Flavonoids, Tannins,

Phenolic compounds, Steroids, Carbohydrates Alkaloids Proteins, Amino acids,

Saponins, Fixed oils and Volatile oils

Table.10 - Preliminary Phytochemical analysis of Canavalia gladiata seed

Tests Alcoholic

extract

Pet.

Ether

Ethyl

acetate

H2O

Carbohydrate + - - +

Alkaloids + + -

Sterols + + - -

Flavonoids + - + +

Tannins + - + +

Saponins + - - +

Fixed oils and fats - + - -

Proteins and Amino acids + - - +

Volatile oils - + - -

+ = Present , - = absent

Macroscopic evaluation:

Colour : Seeds are white, Shiny,

Shape : Seeds are compressed, ellipsoid

Size : 2.5 cm long, 2.4 mm wide

Odour : Characteristic

Taste : Slightly bitter



Microscopic evaluation:

Microscopy of Canavalia gladiata seed powder, study revealed presence of

endosperm cells with aleurone grains, big ellipsoidal oil globules, starch grains, pinkish

brown coloured stone cells.

Ultra – Violet analysis:

Powdered drug under Ultra Violet and ordinary light when treated with different

reagents emitted various colour radiations which helped in identifying the drug in

powder form.

Treatment Visible light UV light

(365 nm)

Powder as such White No fluorescence

1M HCl Colourless No fluorescence

Picric acid Yellow Parrot green fluorescence

50% H2SO4 Dull brown Pale brownish fluorescence

50% HNO3 Yellow Green fluorescence

Acetic acid Colour less ppt No fluorescence

Fecl3 Blakish brown Dark green fluorescence

Iodine Bluish brown Dark green fluorescence

Methanolic NaOH Light brown Water green fluorescence

Ethanolic NaOH Pale yellow Light green fluorescence

Dragendroffs Reddish brown No fluorescence

Ninhydrine Violate Light purple fluorescence

Canavalia gladiata – Discussion


Quantitative Pharmacognostic Evaluation:

Determination of Ash Content:

Total Ash: Total ash content of Canavalia gladiata seed powder was found to be 3.8%.

The maximum acid insoluble ash was found to be 0.14% where as water soluble ash

was found to be 0.22%.

Loss on drying:

The moisture content of Canavalia gladiata seed powder was found to be 1.9%.

Discussion:

In the present study, Alcoholic extract of (CG) seed was investigated for its

effect on the central nervous system. The plant extract possessed CNS stimulant

activity as indicated by the significant increased alertness, motor coordination,

spontaneous motor activity, climbing and swimming time in FST and decreased

immobility time in tail suspension test and forced swimming test. These findings

suggest that alcoholic extract of CG significantly increased motor activity; it could be

attributed to the antidepressant effect of the extract. The extract also increased the time

spent on the rotarod test, a test mainly used to screen centrally acting muscle relaxants

(Rakotonirina, 2001). Hole board test evaluates the effect of extract on alertness. FST

and TST models of depression are widely used to screen new antidepressant drug

(Porsolt et al, 1977; 1979; Steru et al, 1985). These tests are quite sensitive and

relatively specific to all major classes of antidepressant drug (Porsolt et al, 1977; Steru

et al, 1985; Detke et al, 1995). Extract of CG decreased the immobility in FST and TST

indicates CNS stimulant effect. The alcoholic extract may have active constituents with

CNS stimulant activity.



MPTP caused significant overt and subtle behavioural manifestations. Present

study reveals the existence of quantitative behavioural responses with in a short span of

seven days in MPTP lesioned animals with the administration of CG in a dose

dependent manner. Present study evaluated on 1st, 3

rd and 6

th day of treatment. MPTP

administered mice, subjected to the rotarod test, revealed a significant loss of muscular

coordination, it could be due to loss of muscular strength as reported earlier (Tillerson

et al, 2002). Reduced locomotor activity, could be due to depressant action. The

alcoholic extract of Canavalia gladiata prevented the motor impairment in a dose

dependent manner which was altered by MPTP. Maximum effect of the extract was

observed on 6th

day of treatment. Loss of brain dopamine was observed in MPTP

treatment animals. An altered behavioural responses in mice, followed by DA depletion

is considered to be similar to human Parkinsonism (Cousins et al, 1996; Pisa, 1998;

German et al, 1996).

The reduction of brain dopamine and other amines like norepinephrine,

epinephrine, and serotonin were observed in MPTP treated groups. Dopamine was

much affected in Parkinson’s disease but other monoamines level were much less

affected than dopamine (Robert et al, 1987), because main targeted neurotransmitter is

dopamine in Parkinson’s disease. These findings are near to previously reported study

(Horykiewicz, 1960; Kish, 1986).

Norepinephrine was slightly altered in Parkinsonian patient and their level was

significantly below normal this is in accordance with previously reported reduced nor

epinephrine level in brain of the Parkinson’s patient (Stephen et al, 1984). This

neurotransmitter has been related to attention and learning altered norepinephrine



metabolism may contribute to some aspects of intellectual dysfunction in PD (Yadav

et al, 1984).

Dopamine is the precursor to norepinephrine, norepinephrine is a precursor to

the harmone epinephrine. Norepinephrine and epinephrine are antistress chemicals in

the body, obviously there is great stress from parkinsons disease. Further, epinephrine

is involved in increasing the power of muscles and prolonging the action of muscle, by

its ability to activate the release of glucose from glycogen. Thus optimizing the ability

of epinephrine may help achieve more muscle control, perhaps reducing motor

symptoms of parkinson’s (Bonnet and Houeto, 1999).

Serotonin content was slightly altered in MPTP treated group. This statement is

similar to earlier reports (Robert et al, 1987).

Depression is a common symptoms in patients with parkinsons disease.

Alterations in serotonin metabolism are found in primary depression. The brain content

of serotonin in parkinsons disease is also reduced, but this has not bean related to any

manifestation of the disorder. Cerebrospinal fluid (CSF) content of the major major

metabolite of serotonin, 5-hydroxyindoleacetic acid was lower in depressed than

nondepressed parkinsonians. The data suggest that the alterations in serotonin

metabolism in parkinsons disease identify a subgroup of patients who prone to

depression (Mayeux et al, 1984). DA and 5-HT transporters are differentially affected

in PD and 5-HT transporters in the mid brain region may not be affected in relatively

early stages of PD (Sang et al, 2003).

The loss of brain dopamine and other amines was increased by various fractions

of CG in dose dependent manner but maximum neuroprotection was observed with

alcoholic and water extract. It could be due to presence of L-dopa, Flavonoids and



Polyphenols present in CG seed extract. L-DOPA could be obtained in good yield on

extraction with ethanol and water (Laxminarain et al, 2007). These compounds might

cross the blood brain barrier. Pet. ether fraction of CG had minimum effect. It might

suggest that some component other than L-Dopa might also be responsible for

antiparkinsonian activity (Laxminarain et al, 2007). Further, it might be due to presence

of other compounds in addition to L-DOPA, (or) adjuvants responsible to enhance the

efficacy of L-DOPA (Hussain et al, 1997).

Mechanism of neuroprotective action of MPTP involves a cascade of events

beginning with the dopamine transporter which is thought to play an important role in

the uptake of its neurotoxic metabolite MPP+ (Javitch et al, 1985), possibly accounting

for its selectively as a toxin for dopaminergic neurons. The neurotoxic effect of MPTP

can be blocked following treatment with dopamine uptake blocker’s (Giros and Caron,

1993) or reduction in dopamine transport expression (Gainetdinov et al, 1997; Van

kampen et al, 2000). Certain fractions of CG have been shown to inhibit the uptake of

dopamine in to mice synaptosomes (Tsang et al, 1985) these findings provide primary

evidence for its potential as a clinically meaningful antiparkinsonian medication,

including PD prevention, improvement of PD symptoms and adjunctive therapy with L-

DOPA and its related compounds present in it. Present findings are at least in part

similar to the previously reported study (Mohana sundari, 2006), therefore the present

study strongly indicates that CG extract may protect against the MPTP induced

decrease in dopamine and other amine levels in mice and this model provides an

avenue to evaluate many antiparkinsonian compounds.

Lipid peroxidation is a complex process occurring in aerobic cells and reflects

the interaction between the molecular oxygen and polyunsaturated fatty acids. This



involves formation and propagation of lipid alkoxy, lipid peroxy radicals, lipid

hydroxyperoxides as well as variety of degradation products (Gutteridge and Halliwell,

1990). By products of lipid peroxidation cause marked alteration in the structural integrity

and function of cell membranes. Lipid peroxidation by products formed under

physiological and pathological conditions are scavenged by enzymatic and nonenzymatic

antioxidants.

An imbalance between antioxidant defence mechanism and lipidperoxidation

processes results in cell and tissue damage (Yuksel et al, 2001). Lipid peroxidation was

increased in MPTP induced Parkinson’s mice that was reversed by CG seed extract

(possibly due to its antioxidant potential as observed by increased reduced glutathione

and decreased lipid peroxidation level). It could be due to presence of higher

concentration of flavonoids and other compounds in CG. These results were in

accordance with earlier reports of (Silva et al, 2005).

Glutathione, as a major antioxidant in the brain, plays a key role in defending

against oxidative damage (Shanti and Dandiya 1994, Cui et al, 2006). GSH is a tripeptide

comprised of glutamate, cysteine and glycine, where it functions as antioxidant protecting

cells from toxic effects of ROS (Halliwell et al, 1989). In the present study, reduced

glutathione activity was decreased in MPTP induced Parkinson’s mice. It was increased

with all the fractions of CG in dose dependent manner, indicating that CG has an

antioxidant activity.

In the present study various invitro antioxidant studies were also carried out to

asses scavenging potential of CG extract. The DPPH test provides information on the

reactivity of the extract with stable free radicals. DPPH is stable nitrogen centered free

radical containing an odd electron in its structure that can accept an electron or



hydrogen radical to become a stable diamagnetic molecule or hydrogen radical to

become a stable and usually utilized for detection of radical scavenging activity (Blois

et al, 1958). DPPH radicals react with suitable reducing agent as a result of which the

electrons become paired off forming the corresponding hydrazine. The solution

therefore loses colour Stoichometrically depending on the number of electrons uptake

(Blois, 2001). From the result of the present study, it may be postulated that active

principle in Canavalia gladiata seed has hydrogen donors thus scavenging the free

radical. The results are similar to earlier reports observed with origanum majoram

(Mangathayaru et al, 2007).

The alcoholic extract of CG effectively inhibited the lipid peroxidation in a

different dose concentrated manner. This activity is perhaps related to the H+ ions

donating capability of the extract, which scavenges the peroxyl radical to inhibit or

terminate of the peroxidation chain (Ohkowa et al, 1979).

Nitric oxide (NO) is an important chemical mediator generated by the

endothelical cells, neurons involved in the regulation of various physiological processes

(Sainani et al, 1997). Oxygen reacts with the excess NO to generate nitrite and

peroxynitrite anions, which acts as free radicals. In the present study, the nitrite

produced by the incubation of solution of sodium nitroprusside in standard phosphate

buffer at 25° was reduced by alcoholic extract of CG. This may be due to the

antioxidant principles in the Canavalia gladiata extracts, which compete with oxygen to

react with nitric oxide there by inhibiting the generation of nitrite (Marcocci et al,

1994).

Superoxide dismutase catalyses the dismutation of the highly reactive super

oxide anion to oxygen and hydrogen peroxide (Kamalakkannan et al, 2003). Super

oxide anion is the first reduction product of oxygen (Ray et al, 2002).



Preliminary phytochemical analysis indicated the abundance of true alkaloids

and flavonoids in CG seed extract. The antioxidant potential could be attributable to

these putative constituents. Flavonoids are well known natural anti oxidantants due to

this electron donating property which either scavenge the principal propagating free

radicals can hold the radical chain(Sugihara et al,1999).

Pharmacognostical study of the seed of CG was carried out in order to identify

the correct identification of this plant and to differentiate closely related other species

of Cannavalia. Data obtained would serve as standard reference for identification of

Canavalia gladiata.

Conclusion:

Canavalia gladiata exhibited significant antiparkinsonian activity in MPTP

mouse model. It appears to be the most promising plant due to its l-dopa content and

potential antioxidant activity. The probable mode of action of this plant may be due to

the increased in DA content by CG and may be due to increased synthesis or decreased

metabolism. The results indicate increased DA synthesis may be contributed for

antiparkinsonian activity. These findings provide preliminary evidence for its potential

as a clinically meaningful antiparkinsonian medication, including Parkinsonian disease

prevention, improvement of Parkinson’s disease symptoms. Further studies are required

to explore the chemical constituents responsible for the activity and also to establish the

mode of action.

Barleria prionities - Results


BARLERIA PRIONITIES

Results of Pharmacological studies:

Acute toxicity and gross behaviour changes:

Alcoholic extracts and its fractions of Barleria prionities found to be safe since

no animal died even at the maximum single dose of 3200 mg/kg orally. The animals

did not show any gross behavioural changes at the doses tested.

Evaluation of Neuropharmacological studies:

The animals treated with a dose of 100, 200, 300 mg/kg of alcoholic extract of

Barleria prionities (BP) root showed significant (P<0.05) increase of spontaneous

motor activity at a dose of 400 mg/kg as compared to control vehicle group (Fig.40).

Alcoholic extract of BP at given doses showed no significant change in reaction

time as compared to control group in Eddys hot plate (Fig.41)

The results of Rota rod test revealed that there was significant (P<0.01) dose

dependent increase of motor coordination at a dose of 200, 300 mg/kg of BP extract as

compared to control vehicle group (Fig.42)

Alcoholic extract of BP at given doses significantly (P<0.01) dose dependently

decreased immobility time in Tail suspension at a dose of 200, 300 mg/kg as compared

to control vehicle group (Fig.43).

At given doses alcoholic extract of BP showed significant (P<0.01) dose

dependent decrease in immobility time and significant (P<0.01) increase in swimming

time in forced swimming test as compared to control vehicle group (Fig.44 & 45).



Animals treated with a dose of 100, 200, 300 mg/kg alcoholic extract of BP

significantly (P<0.05, 0.001) increased in the number of head dipping at a dose of 200

& 400 mg/kg as compared to control vehicle group (Fig.46).

Effect on behaviour of animals after MPTP treatment:

Spontaneous motor activity significant decrease (P<0.001) was observed in MPTP

treated group as compared to control group. Locomotor activity was significantly

(P<0.001) increased with alcoholic extract of BP at higher dose on 1st day but it was

significantly (P<0.001) dose dependently increased on 3rd

and 6th

day of treatment as


Results of Rota rod test showed significant (P<0.001) decrease of motor co-

ordination in MPTP treated group as compared to control group. Retention time was

reversed significantly (P<0.001) dose dependently on 1st, 3

rd and 6

th day of treatment

with alcoholic extract of BP root as compared to MPTP treated group (Table.11;

Fig.48).

Animals treated with MPTP significantly (P<0.001) reduced the alertness as

compared to control vehicle group. Alertness was significantly (P<0.001) increased

with alcoholic extract of BP at a dose of 200 & 300 mg/kg on 1st day and it was

significantly (P<0.001) dose dependently increased on 3rd

and 6th

day of treatment as


Antiparkinsonian Activity:

Effect on dopamine level in the brain:

The result of the present study revealed the dopamine level was significantly

(P<0.001) decreased in the MPTP treated animals as compared to control group and



their level was significantly (P<0.001) increased at 200 & 300 mg/kg with alcoholic

extract of BP (AEBP) as compared to MPTP group (Table.12; Fig.50).

The dopamine level was not significantly altered at a dose of 100 mg/kg but it

was significantly increased at higher doses with petroleum ether extract of Barleria

prionities (PEEBP) as compared to MPTP treated group (Table.12; Fig.51).

The ethyl acetate extract of Barleria prionities (EAEBP) showed significant

(P<0.001) dose dependent increase of dopamine level at higher doses as compared to

lower doses and MPTP treated group (Table.12; Fig.52).

The dopamine level was significantly (P<0.001) dose dependently increased

with water extract of BP (WEBP) as compared to MPTP group (Table.12; Fig.53).

Effect on brain epinephrine level:

Epinephrine level was significantly (P<0.001) decreased in MPTP treated group

as compared to control vehicle group. While their level was significantly (P<0.001)

increased with AEBP at 300mg/kg as compared to MPTP group (Table.13; Fig.54)

Significantly (P<0.001) increased of epinephrine level with PEEBP, EAEBP

andWEBP at a dose of 300 mg/kg as compared to MPTP group (Table.13; Fig.55, 56,

57).

Effect on brain norepinephrine level:

Results of the present study showed that norepinephrine level was significantly

(P<0.001) altered by MPTP treated group. While it was significantly increased at a

dose of 300 mg/kg of alcoholic extract of BP as compared to MPTP group. NE level

was not significantly increased with pet. ether fraction in combination with MPTP but it

was significantly (P<0.001) increased with plant extract alone at a dose of 300 mg/kg

as compared to MPTP treated group (Table.14; Fig. 58 and 59).



Treatment with EAEBP and WEBP significant (P<0.001) increase of

norepinephrine level was observed at higher dose as compared to MPTP treated

animals (Table.14; Fig.60 and 61).

Effect on brain serotonin level:

The serotonin level was significantly (P<0.001) reduced in MPTP treated mice

while its level was restored significantly (P<0.01) at a dose of 200, 300 mg/kg of AEBP

as compared to MPTP group (Table.15; Fig.62).

Animals treated with EAEBP and PEEBP showed significant (P<0.001)

improvement at a dose of 300 mg/kg plant extract alone as compared to MPTP group

(Table.15; Fig.63 and 64).

Water extract of BP exhibited significant (P<0.001) increase of serotonin level

at 200 mg and 300 mg/kg as compared to MPTP group (Table.15; Fig.65).

Invivo antioxidant studies:

Effect on Lipid peroxidation:

Result of the present study showed that brain MDA level was significantly

(P<0.001) increased in the MPTP treated group as compared to control vehicle group.

On treatment with alcoholic extract of BP significantly (P<0.001) dose dependently

decrease of MDA level was observed as compared to MPTP treated animals. Pet. ether

extract of BP exhibited significant (P<0.01) decrease of MDA level at a dose of 300

mg/kg as compared to MPTP treated group (Table.16; Fig. 66 and 67).

EAEBP showed significant (P<0.001) reduction of TBARS level at a dose of

200& 300 mg/kg as compared to low dose and MPTP treated group (Table.16; Fig. 68)



Animals treated with WEBP in combination with MPTP revealed significant

(P<0.001) decrease of TBARS level at higher dose as compared to MPTP group

(Table.16; Fig.69).

Effect on Reduced glutathione level:

Reduced glutathione level was significantly (P<0.001) reduced in MPTP treated

group as compared to control group. GSH level was restored significantly (P<0.05,

0.01) at a dose of 200, 300 mg/kg with alcoholic extract of BP as compared to MPTP


PEEBP given in combination with MPTP showed moderately significant

(P<0.05) increase of GSH level at 300 mg/kg as compared to MPTP treated group

(Table.17; Fig.71).

EAEBP exhibited significant (P<0.01) effect on GSH level at 300 mg/kg as

compared to MPTP treated animals (Table.17; Fig.72).

WEBP showed significantly (P<0.05, 0.001) enhanced the GSH level at a dose

of 200, 300 mg/kg as compared to MPTP treated group (Table.17; Fig.73).


Institute of Pharmaceutical Technology, SPMVV, Tirupati.

104

Table. 11: Effect of Alcoholic extracts of Barleria prionities root on Spontaneous motor activity, Grip strength, Alertness (Hole Board

test) in MPTP treated mice

Groups Spontaneous motor activity score

Grip Strength in Seconds Alertness (no. of head dippings)

1st day 3

rd day 6

th day

1st day 3

rd day 6

th day 1

st day 3

rd day 6

th day

Control 446.6 ± 12.0

446.6 ± 12.0 446.6 ± 12.0 120 ± 0 120 ± 0 120 ± 0 50.5 ± 2.23 50.5 ± 2.23 50.5 ± 2.23

MPTP 278.5 ±0.7

***

179.8 ± 5.3

***

90.3 ± 6.2

***

83 ± 2.8

***

33.6 ± 7.2

***

13.8 ± 1.6

***

22.3 ± 1.6

***

14.0 ± 2.1

***

8.6 ± 1.2

***

100mg/kg CGE

+

MPTP

285.8 ± 5.6

***

292.6 ± 2.4

***, +++

300.6 ± 1.3

***, +++

92 ± 3.0

***, +

91.6 ± 1.7

***, +++

95.83 ± 1.0

***, +++

24.8 ± 1.5

***

31.5 ± 1.2

***, +++

34.0 ± 1.7

***, +++

200 mg/kg CGE

+ MPTP

299.1 ± 1.1

***

302.6 ± 1.3

***, +++

324.0 ± 3.5

***, +++

96 ± 1.6

***, ++

98.5 ± 0.9

***, +++

102.1 ± 3.3

***, +++

31.0 ± 1.1

***, ++

35.8 ± 1.1

***, +++

37.5 ± 0.5

***, +++

300 mg/kg CGE

+ MPTP

303.1 ± 1.8

***, +++

328.1 ± 3.1

***, +++

436.8 ± 2.3

+++

100 ± 1.0

***, +++

101 ± 0.7

**, +++

14.3 ± 1.4

***, +++

31.6 ± 1.1

***, ++

38.6 ± 1.2

***, +++

39.6 ± 1.1

***, +++

300 mg/kg CGE 317.0 ± 3.1

***, +++

336.8 ± 3.0

***, +++

455.5 ± 17.4

+++

101 ± 2.2

***, +++

106.8 ± 1.4

+++

111.6 ± 0.8

**, +++

46.8 ± 1.6

+++

48.6 ± 1.0

+++

52.0 ± 1.8

+++


** (P<0.01), *** (P< 0.001) Vs Control group; + (P< 0.05), ++ (P< 0.01), +++ (P< 0.001) Vs MPTP group.



105

Table.12: Effect of Barleria prionities root on brain dopamine level in MPTP pretreated mice.

Groups

g/g brain tissue


Control 3.815 ± 0.055 3.815 ± 0.055 3.815 ± 0.55 3.815 ± 0.055

MPTP 1.257 ± 0.038*** 1.257 ± 0.038*** 1.257 ± 0.038*** 1.257 ± 0.038***

100mg/kg BPE+ MPTP 1.516 ± 0.12*** 1.280 ± 0.042*** 1.317 ± 059*** 1.399 ±0.049***

200 mg/kg BPE + MPTP 1.926 ± 0.11***

+++

1.585 ± 0.044***

++

1.895± 0.14***

++

1.901 ± 0.036***

+++

300 mg/kg BPE + MPTP 2.915 ± 0.09 ***

+++

2.465 ± 0.093***

+++

2.875 ± 0.050***

+++

2.922 ± 0.032***

+++

300 mg/kg BPE 3.088 ± 0.12***

+++

2.749 ± 0.039***

+++

3.006 ± 0.024***

+++

3.015 ± 0.029***

+++


*** (P<0.001) Vs Control group;

++ (P< 0.01), +++ (P< 0.001) Vs MPTP group.



106

Table.13: Effect of Barleria prionities root on brain epinephrine level in MPTP treated mice.

Groups

g/g brain tissue


Control 3.288 ± 0.102 3.288 ± 0.102 3.288 ± 0.102 3.288 ± 0.102

MPTP 2.106± 0.090*** 2.106 ± 0.090*** 2.106 ± 0.090*** 2.106 ± 0.090***

100mg/kg BPE + MPTP 2.213± 0.068*** 2.131 ± 0.154*** 2.151 ± 0.46*** 2.192 ± 0.079***

200 mg/kg BPE + MPTP 2.264 ± 0.062***

2.198 ± 0.104*** 2.116 ± 0.111***

2.251 ± 0.040***

300 mg/kg BPE + MPTP 2.635 ± 0.045***

+++

2.214 ±0.038***

2.670 ± 0.038 ***,

+++

2.738 ± 0.043 ***

+++

300 mg/kg BPE 3.208 ± 0.137+++ 2.913 ± 0.054+++ 3.035 ± 0.065+++ 3.100 ± 0.083+++



+++ (P<0.001) Vs MPTP group.



107

Table.14: Effect of Barleria prionities root on brain norepinephrine level in MPTP pretreated mice.

Groups

g/g brain tissue

Alcoholic Pet-ether Ethyl acetate Water extract

Control 3.343 ± 0.054 3.343 ± 0.054 3.343 ± 0.054 3.343 ± 0.054

MPTP 2.148 ± 0.041 *** 2.148 ± 0.41 *** 2.148 ± 0.041 *** 2.148 ± 0.041 ***

100mg/kg BPE + MPTP 2.214 ± 0.070 *** 2.164 ± 0.071*** 2.168 ± 0.046 *** 2.190 ± 0.037 ***

200 mg/kg BPE + MPTP 2.289 ± 0.125 *** 2.170 ± 0.082 ***

2.184 0.054 ***

2.205 ± 0.054 ***

300 mg/kg BPE + MPTP 2.850 ± 0.044 ***

+++

2.202 ± 0.037 ***

+++

2.703 ± 0.088 ***

+++

2.731 ± 0.083 **

+++

300 mg/kg BPE 3.293 ± 0.157 +++ 3.028 ± 0.046 ** 3.158 ± 0.058+++ 3.173 ± 0.055 +++






108

Table.15: Effect of Barleria prionities root on brain serotonin level in MPTP treated mice.

Groups

g/g brain tissue


Control 3.407 ± 0.050 3.407 ± 0.050 3.407 ± 0.050 3.407 ± 0.050

MPTP 2.068 ± 0.060 *** 2.068 ± 0.060 *** 2.068 ± 0.060 *** 2.068 ± 0.060 ***

100mg/kg BPE + MPTP 2.411 ± 0.091 *** 2.125 ± 0.047 *** 2.219 ± 0.162 *** 2.221 ± 0.085 ***

200 mg/kg BPE + MPTP 2.508 ±0.284 ** 2.227 ± 0.100 *** 2.323 ± 0.047 ***

2.469 ± 0.119 ***

+

300 mg/kg BPE + MPTP 2.616 ± 0.084 **

2.344 ± 0.068 ***

2.783 ± 0.04 ***

+++

2.515 ± 0.057 ***

++

300 mg/kg BPE 3.321 ± 0.134 +++ 2.969 ± 0.090 **

+++

3.038 ±0.035 *

+++

3.106 ± 0.055 +++


* (P< 0.05), ** ( P<0.01), *** (P< 0.001) Vs Control group ;

+ (P< 0.05), ++ (P< 0.01), +++ (P< 0.001) Vs MPTP group.



109

Table.16: Effect of Barleria prionities root on Lipid peroxidation (nmol/mg protein/hr) level in MPTP treated mice.

Groups

nmol/mg protein/hr


Control 26.25 ± 3.4 26.25 ± 3.4 26.25 ± 3.4 26.25 ± 3.4

MPTP 67.0 ± 5.6 *** 67.0 ± 5.6 *** 67.0 ± 5.6 *** 67.0 ± 5.6 ***

100mg/kg BPE + MPTP 44.83 ± 3.3 **

+++

61.58 ± 1.9 *** 59.20 ± 2.2 *** 56.18 ± 2.2 ***

200 mg/kg BPE + MPTP 33.5 ± 1.9 +++ 56.68 ± 2.2 *** 45.25 ± 3.8 **

+++

43.83 ± 3.0 **

+++

300 mg/kg BPE + MPTP 27.3 ± 1.4 +++ 45.00 ± 3.3 *

++

35.29 ± 2.8 +++ 35.29 ± 2.1 +++

300 mg/kg BPE 22.7 ± 2.1 +++ 31.83 ± 1.6 +++ 29.55 ± 0.5 +++ 28.02± 0.8 +++


* (P< 0.05), ** (P< 0.01), *** (P< 0.001) Vs Control group;

++ (P< 0.01), +++ (P< 0.001) Vs MPTP group.



110

Table. 17: Effect of Barleria prionities root on reduced glutathione (µmol/mg protein) level in MPTP pretreated mice.

Groups

µmol/mg protein


Control 759.90 ± 53.2 759.90 ± 53.2 759.90 ± 53.2 759.90 ± 53.2

MPTP 476.60 ± 24.9 *** 476.60 ± 24.9 *** 476.60 ± 24.9 *** 476.60 ± 24.9 ***

100mg/kg BPE + MPTP 544.33 ± 24.9 *** 484.43 ± 22.1 *** 496.4 ± 4.6 *** 542.87 ± 18.6 ***

200 mg/kg BPE + MPTP 616.73 ± 16.5 *

+

494.43 ± 6.1 *** 548.97 ± 20.9 *** 609.17 ± 13.2 **

+

300 mg/kg BPE + MPTP 676.10 ± 15.8 ++ 597.90 ± 6.5 **

+

631.35 ± 16.4 *

++

666.10 ± 16.3 +++

300 mg/kg BPE 721.17 ± 20.9 +++ 709.20 ± 15.1 +++ 729.17 ± 4.8 +++ 729.53 ± 4.6 +++



+ (P< 0.05), ++ (P<0.01), +++ (P< 0.001) Vs MPTP group.



Invitro antioxidant Studies:

DPPH:

The antioxidant property of AEBP was evaluated by DPPH radical scavenging

assay. IC50 value of AEBP was 360 µg/ml these results are given in (Table.18;

Fig.74).

Lipid Peroxidation:

The antioxidant property of AEBP was also evaluated by the inhibition of

malondialdehyde formation generated by Fe2+

ascorbate in rat brain homogenate. The

IC50 value of AEBP was 380 µg/ml (Table.18; Fig.75).

Nitric Oxide (NO) Scavenging:

The Alcoholic extract of BP showed promising free radical scavenging action

against NO induced release of free radical with IC50 value of AEBP was found to be

at390 µg/ml (Table.18; Fig.76).

Super Oxide dismutase:

AEBP showed the inhibition of free radical scavenging activity with IC50 value

of 400 µg/ml (Table.18; Fig.77).



Table.18 Effect of alcoholic extracts of Barleria prionities root on different

antioxidant models.

S.No.

% inhibition

Conc

µg/ml

DPPH Superoxide

dismutase

Nitric

oxide

Lipid

Peroxidation

1 3 6.83 12.19 9.62 17.4

2 6 11.29 18.5 18.6 19.8

3 12 18.96 24.5 20.2 22.4

4 25 23.7 26.8 25.5 27.7

5 50 39.4 39.34 30.4 31.6

6 100 47.8 42.79 36.8 36.4

7 200 48.2 49.59 42.3 45.7

8 400 54.8 53.6 52.1 54.9

9 800 87.9 78.6 86 87.6

10 IC50 (µg/ml) 360 400 390 380




Results of Pharmacognostic evaluation:

Table.19 Phytochemical analysis of Bareleria prionities extract showed presence

of following phytoconstituents.

Tests Alcoholic Pet. ether Ethyl acetate Water

Flavonoids + - - +

Tannins and Phenolic + - - + +

Steroids - + - -

Carbohydrates - - - -

Alkaloids + - + -

Protein & amino acids + - - +

Saponins + - - +

Fats - + - -

Volatile oils - - - -

Anthraquinone glycosides + - - +

Glycosides + - - -

+ = present , - = absent


Colour - Roots are light yellow

Shape - Cylindrical, slight curved, hairy

Odour - Acceptable, characteristic

Taste - Characteristic


Microscopic analysis of BP root powder showed the presence of polygonal with

brown colour cork cells, prismatic calcium oxalate crystals, lignified stone cells, xylem

vessels, sharp end phloem fibers and oval shaped compound and simple (rare) blue

coloured starch grains.



Behaviour of BP root powder with different reagents:

Reaction Colour observed Colour observed

Under visible light under UV light

(365 nm)

Powder as such light brown No fluorescence

Powder + Picric acid Yellowish brown Bright green fluorescence

Powder + HCl Mud colour light blue fluorescence

Powder + H2SO4 Honey Parrot green fluorescence

Powder + HNO3 Pinkish brown water green fluorescence

Powder + glacial acetic acid Colourless light green fluorescence

Powder + FeCl3 light brown Dark green fluorescence

Powder + iodine Dark reddish brown green fluorescence

Powder + NaOH (aq) Light yellow parrot green fluorescence

Powder + NaOH (Methanolic) Cream parrot green fluorescence

Powder + HCl + H2O Colourless No fluorescence

Powder + H2SO4 + H2O Colourless Light blue fluorescence

Powder + HNO3 + H2O Colourless Light copper sulphate fluorescence

Powder + BaCl2 White colour Sky blue fluorescence

Results of Quantitative powder analysis:

Total ash:

Total ash content of Barleria prionities root was found to be 6% where as Acid

insoluble ash and water soluble ash was found to be 2% and 3.2% respectively.

Moisture content:

Moisture content of Barleria prionities root powder was found to be 1.6%.

Barleria prionities - Discussion


Discussion:

Results of the alcoholic extract of Barleria prionities suggested that the

alcoholic extract possess central nervous system stimulant activity with the significant

increase of locomotor activity, alertness, retention time, swimming time in FST. And

significantly decreased immobility time in TST and FST. These evidences suggest that

Barleria prionities root could contain CNS stimulant active constituents. Antidepressant

effect of the extract could be evaluated by FST & TST. These test are quite sensitive

and widely used to screen antidepressant drugs (Porsolt et al, 1977, Steru et al, 1985;

Detke et al, 1995).

Alcoholic extract of BP also increased the retention time in Rota rod test. The

extract could have muscle coordination which might be due to presence of phyto

constituent with CNS stimulant action (Rakotonirina, 2001).

The study also revealed the quantitative behavioural responses with in a short

span of seven days in MPTP lesioned animals. The treated animals were subjected to

the spontaneous motor activity, rota rod test, and exploratory behaviour. Decreased

SMA could be due to motor impairment, retention time was also decreased it could be

due to loss of muscular strength. Number of head dippings were also reduced. It might

be due to motor impairment. Behavioral complication altered by MPTP treatment was

reversed with alcoholic extract of BP root. Maximum protection of extract on 6th

day of

treatment might be due to lag time required for restoration.

Dopamine depletion was also occurred in MPTP treated mice. Altered

behavioral responses followed by DA depletion are similar to human Parkinsonism

(Cousins et al 1996; Pisa, 1998; German et al, 1996).



Dopamine neurotransmitter was more affected in Parkinson’s disease

(Horykiewicz, 1960 and Kish, 1986) where as other brain amines like norepinephrine,

epinephrine and serotonin were much less affected than dopamine in MPTP treated

group (Robert et al, 1987). Norepinephrine was less altered in parkinsonian patient this

is in accordence with earlier reported studies (Stephen et al, 1984). Altered

norepinephrine might contribute to some aspects of intellectual dysfunction in PD

(Yaadov et al, 1984).



the body, obviously there is great stress from parkinsons disease. Further, epinephrine





MPTP treated group showed slight alteration of serotonin level, it is similar to

earlier report (Robert et al, 1987).

Depression is a common symptoms in patients with parkinsons disease.


of serotonin in Parkinson’s disease is also reduced, but this has not bean related to any


metabolite of serotonin, 5- hydroxyindoleacetic acid was lower in depressed than

nondepressed parkinsonian’s. The data suggest that the alterations in serotonin

metabolism in parkinson’s disease identify a subgroup of patients who prone to

depression (Mayeux et al, 1984). DA and 5-HT transporters are differentially affected



in PD and 5-HT transporters in the mid brain region may not be affected in relatively

early stages of PD (Sang et al, 2003).

The loss of dopamine and other amines was enhanced by all the fractions of BP

in a dose dependent manner. Among all the fractions alcoholic and water fractions

showed maximum neuroprotection. It could be due to presence of more water soluble

components like flavonoids, proteins and L-dopa. Alcohol and water are good solvents

for extraction of L-dopa (Laximinarain et al, 2007). Antiparkinsonian effect of BP

extract not only due to presence of L-dopa but also other compounds, due to this reason

pet. ether fraction of BP might have minimum antiparkinsonian effect. Phytochemical

analysis of pet.ether fraction showed presence of fats and steroids. Fat might act as

adjuvant to increase the efficacy of L-dopa (Hussain et al, 1997). Reactive species can

be eliminated by a enzymatic and non enzymatic antioxidants and thus protecting

tissue/ organ damage from oxidative stress. In the present study, we estimated non

enzymatic antioxidant and brain lipid peroxidation in vivo.

Malondialdehyde (MDA) is an end product of lipidperoxidation, a non

enzymatic antioxidant present in less concentration scavenges hydroxyl free radicals

(Auddy et al, 2003). In our study, increase of MDA level was observed in MPTP

treated mice than in the group treated with plant extract. These findings are similar to

earlier reports (Amarnath, 2004) on Boerhavia diffusia Linn. MDA level was decreased

with all the fractions of BP root. Glutathione is an nonenzymatic antioxidant.

Glutathione peroxides a selenium containing enzyme present in significant concentrations

detoxifies H2O2 to H2O through the oxidation of reduced glutathione (Bruce et al,

1982). The reduced glutathions level was observed in MPTP treated mice. GSH level

was restored with various fractions of root. Among all fractions alcoholic and water



extract was more significant. Medicinal herbs with antioxidants are useful in diseases in

which free radicals are involved such as, anoxia, ischemia of brain neuronal cell death

in Parkinson’s, arteriosclerosis, rheumatism and cancer (El-Tahir et al, 1993; Houghton

et al, 1995; Medicana et al, 1997; Badary et al, 2000; Al- Ghamdi, 2001). Several

studies on medicinal plants with free radical scavenging and antioxidant activities

indicates that these activities are due to presence of polyphenols and flavonoids (Lu and

FOO, 2001; Damintoti et al,2005; Ivanova et al, 2005). Earlier chemical studies (Rao

and Raju, 1979; 1981; 1984; Sankaranarayanan et al, 2010; Pettes et al, 1981; Bharati

sinha et al, 1982; Harborne et al, 1983; Virinder et al, 1989; Kamal and Mangla, 1993

Narender et al, 2006) and preliminary phytochemical test indicated the presence of

phenolic compounds and flavonoids. Hence the antioxidants and free radical

scavenging activities of the plant may be due to presence of these compounds.

Invitro antioxidant activity of BP was carried out in various antioxidant models.

Oxidative stress has been implicated in the pathology of many diseases (Marx, 1987)

Antioxidants may offer resistance against the oxidative stress by scavenging the free

radicals and by many other mechanisms and thus prevents disease (Youdim et al,

2001).

The DPPH test provides information on the reactivity of test extract with a

stable free radical. DPPH is stable nitrogen centered free radical containing an odd

electron on its structure that can accept an electron or hydrogen radical to become a

stable diamagnetic molecule and usually utilized for detection of radical scavenging

activity (Blois, 1958). Because of its odd electron DPPH gives a strong absorption at

517 nm in the visible region (deep violet colour). As the electron becomes paired off in

presence of a free radical, the absorbance diminishes, thus the resulting decrease in



absorbance is stoichiometric with respect to the number of electrons taken up (Oke

et al, 2002). The AEBP exhibited marked and dose dependent free radical scavenging

effect in DPPH radical scavenging assay showing the IC50 value of 360µg/ml.

Lipid peroxidation can be prevented either by reducing the formation of free

radicals or by supplying the competitive substrate for unsaturated lipids in the

membrane or by accelerating the repair mechanisms of damaged cell membrane.

Several natural and synthetic antioxidants are used to prevent the lipid peroxidation

(Yoshikawa et al, 1983; Valentao et al, 2002).

The antioxidant activity of the AEBP was further confirmed by evaluating

inhibition in production of malondialdehyde (MDA) and related carbonyl products that

are produced as by products of lipid peroxidation induced by Fe2+ ascorbate system in

the biomembrane of rat brain homogenate. These carbonyl products are responsible for

DNA damage, carcinogenesis and aging related diseases (Riemersna et al, 2000). The

MDA reacts with thiobarbituric acid in specific reaction medium to produce a strong

absorption at 532 nm. The AEBP effectively inhibited lipid peroxidation in a dose

related manner exhibiting the IC50 value of 380 µg/ml. this activity is perhaps related

to the H+ ion donating capability of the extract, which scavenges the peroxyl radical to

inhibit (or) terminate the peroxidation chain (Ohkowa et al, 1979).

In the present study the nitrite produced by the incubation of solutions of

sodium nitroprusside in standard phosphate buffer at 25°C was reduced by the

alcoholic extract of BP. This may be due to the antioxidant principles in the extract

which compete with oxygen to react with nitric oxide (Marcocci et al, 1994). There by

inhibiting the generation of nitrite.



Alcoholic extract of BP root scavenges the superoxide radicals. Superoxide

anion is the first reduction product of oxygen (Ray et al, 2002). Which is measured in

terms of inhibition of generation of O2 (Shirwaikar et al, 2004).

Roots of BP are rich in flavonoids. (Flavonoids are natural products which have

been shown to possess various biological properties related to antioxidant mechanism

(Perrisoud et al, 1982).

Pharmacognostic evaluation help in identifying the drug in powder form. Macro

and microscopical characters help in the identification of drug and also in laying down

pharmacopial standards.

Conclusion:

Barleria prionities exhibits dopaminergic neuroprotection in MPTP induced

parkinsonism in mice. It prevents dopaminergic loss and behaviour of animals which are

altered by MPTP. Hence, Barleria prionities can exert a significant neuroprotective effect

and its usefulness in the management of Parkinson’s disease needs exploration.

Prosopis chilensis - Results


PROSOPIS CHILENSIS:

Results of Pharmacological Studies:

Acute toxicity and gross behaviour changes:

Alcoholic extracts and its fractions of Prosopis chilensis was found to be safe

since no animal died even at the maximum single dose of 3200mg/kg orally. The

animal did not show any gross behaviour changes at the doses tested.

Assessment of neuropharmacological activity:

Alcoholic extract of Prosopis chilensis (AEPC) at a given doses significantly

(P<0.05, 0.01) increased spontaneous motor activity at 200 and 300 mg/kg when

compared to control group (Fig. 78).

Animals treated with AEPC significantly (P<0.001) increased the number of head

dippings as compared to control group (Fig. 79).

AEPC exhibited the significant (P<0.01) dose dependent increase of retention

time as compared to control vehicle group (Fig. 80).

Mice treated with AEPC significantly (P<0.01) dose dependently decreased the

immobility time in TST (Fig. 81).

Immobility time was also significantly ((P<0.01) dose dependently reduced and

swimming time was increased at given doses in FST (Fig. 82 and 83).

Antiparkinsonian activity:

Evaluation of behavioural activity in MPTP treated animals:

Results of the present study showed that significantly (P<0.01) decreased

spontaneous motor activity in MPTP treated animals when compared to control group.

AEPC showed no significant effect on 1st day of treatment but locomotor activity



was significantly (P<0.01) dose dependently increased on 3rd

& 6th

day of treatment as

compared to MPTP treated group (Table. 20; Fig. 84).

Retention time was significantly ((P<0.001) decreased in MPTP treated group.

Retention time was significantly (P<0.001) enhanced with AEPC on 3rd

and 6th

of

treatment as compared to MPTP group (Table.20; Fig.85).

Results of the hole board test revealed that the number of head dippings were

significantly (P<0.001) reduced in MPTP treated group as compared to control group.

It was significantly (P<0.001) dose dependently increased on 3rd

and 6th

day of

treatment with AEPC as compared to MPTP treated group (Table. 20; Fig. 86).

Effect on brain dopamine level:

Dopamine level was significantly (P<0.001) reduced in MPTP treated animals

when compared to control group. Its level was significantly (P<0.001) enhanced with

the AEPC at 200 and 300 mg/kg as compared to MPTP group. Animals treated with

Pet. ether extract of Prosopis chilensis (PEEPC) exhibited no significant improvement

when given in combination with MPTP but improvement was observed only with plant

extract (Table. 21; Fig. 87 and 88).

Dopamine level was significantly (P<0.001) increased with the ethylacetate

extract of Prosopis chilensis (EAEPC) at higher dose and same effect was also

observed with water extract of Prosopis chilensis (WEPC) as compared to MPTP

treated group (Table. 21; Fig. 89 and 90).

Effect on brain epinephrine level:

Epinephrine level was significantly (P<0.001) altered in MPTP treated group.

Mice treated with AEPC, EAEPC and WEPC showed significant (P<0.01) enhancement



of epinephrine level at a dose of 300 mg/kg as compared to MPTP group (Table. 22;

Fig. 91, 93 and 94).

PEEPC showed no significant improvement of epinephrine level when compared

to MPTP treated group (Table. 22; Fig. 92).

Effect on brain norepinephrine level:

Norepinephrine level was significantly (P<0.001) changed in MPTP treated

animals as compared to control group. Norepinephrine level was normalized significantly

(P<0.05) with alcoholic extract of Prosopis chilensis as compared to MPTP group

(Table. 23; Fig. 95).

Treatment with PEEPC showed no significant improvement of norepinephrine

level when compared to MPTP treated group (Table. 23; Fig. 96)

Treatment with EAEPC and WEPC exhibited significant (P<0.05) increased the

level of norepinephrine when compared to control group (Table. 23; Fig. 97 and 98)

Effect on brain serotonin level:

Serotonin level was significantly (P<0.001) reduced in MPTP treated mice as

compared to control vehicle group. AEPC in combination with MPTP showed

significant (P<0.001) improvement of serotonin level as compared to MPTP group

(Table. 24; Fig. 99).

Treatment with PEEPC and EAEPC in combination with MPTP did not show

significant effect on serotonin level but when given only plant extract it was effective

significantly (P<0.001) at a higher dose of 300 mg/kg as compared to MPTP treated

group (Table. 24; Fig. 100 and 101).

WEPC exhibited more significant (P<0.001) improvement of serotonin level at

200 and 300 mg/kg when compared with MPTP group (Table. 24; Fig. 102).



Invivo antioxidant studies:

Effect on Lipid peroxidation:

Results of the present study showed that the brain MDA level was significantly

(P<0.001) increased in MPTP treated animals as compared to control group. AEPC

showed significantly (P<0.001) dose dependently reduction of MDA level as compared

to MPTP treated group (Table. 25; Fig.103).

PEEPC showed significant (P<0.01) decrease of MDA level at a dose of 300

mg/kg as compared to MPTP group (Table. 25; Fig.104).

Treatment with EAEPC and WEPC exhibited significant (P<0.01, 0.001) dose

dependent decrease of MDA level at a dose of 200 and 300 mg/kg as compared to

MPTP group (Table. 25; Fig. 105 and 106).

Effect on reduced glutathione level:

Present study showed the reduced glutathione level was significantly decreased

(P<0.001) in MPTP treated mice as compared to control vehicle group. GSH level was

restored significantly (P<0.001) with AEPC as compared to MPTP group (Table.26;

Fig. 107).

Treatment with PEEPC showed no significant effect on GSH level when given

in combination with MPTP but it showed significantly P<0.001) increase of GSH level

only with plant extract alone (Table. 26; Fig. 108).

Both EAEPC and WEPC exhibited significant (P<0.05) increased GSH level at

300 mg/kg as compared to MPTP treated group (Table. 26; Fig. 109 and 110).



Table. 20: Effect of Alcoholic extracts of Prosopis chilensis seed on Spontaneous motor activity, Grip strength, Alertness

(Hole Board test) in MPTP treated mice

Groups Spontaneous motor activity score

Grip Strength in Seconds Alertness (no. of head dippings)

1st day 3

rd day 6

th day

1st day 3

rd day 6

th day 1

st day 3

rd day 6

th day

Control 446.6 ± 12.0 446.6 ± 12.0 446.6 ± 12.0 120 ± 0 120 ± 0 120 ± 0 50.5 ± 2.2 50.5± 2.2 50.5 ± 2.2

MPTP 278.5 ± 7.0

***

179.8 ± 5.3

***

90.3 ± 6.2

***

83.8 ± 2.8

***

33.6 ± 7.2

***

13.8 ± 1.1

***

22.3 ± 1.6

***

14.0 ± 2.1

***

8.6± 1.2

***

100mg/kg CGE

+ MPTP

296.3 ± 1.4

***

290.1 ± 3.6

***, +++

296.1 ± 2.2

***, +++

83.8 ± 1.7

***

89.6 ± 1.6

***, +++

94.0 ± 1.7

***, +++

23.1 ± 1.3

***

28.8 ± 1.0

***, +++

32.0 ± 1.6

***, +++

200 mg/kg CGE

+ MPTP

299± 0.5

***

297.0± 1.8

***, +++

325.0 ± 4.4

***, +++

93.0 ± 2.6

***, +

97.5 ± 0.9

***, +++

99.1 ± 1.0

***, +++

29.8 ± 6.8

***, +

33.5± 0.8

***, +++

34.5 ± 1.3

***, +++

300 mg/kg CGE

+ MPTP

300.8± 2.6

***

326.3 ± 3.1

***, +++

419.6± 7.5

***, +++

97.0 ± 1.5

***, +++

98.8 ± 1.0

***, +++

103 ± 1.0

***, +++

30.8±1.5

***, +

36.1 ± 0.4

***, +++

38.1± 1.8

***, +++

300 mg/kg CGE 326.8 ± 3.6

***,+++

338.8 ± 2.9

***, +++

39.8 ± 7.9

+++

99.8 ± 1.7

***, +++

105.8 ± 1.7

*, +++

109 ± 2.1

***, +++

43.6 ± 2.3

+++

45.8 ± 1.5

+++

49.5± 1.7

+++



+ (P< 0.05), +++ (P< 0.001) Vs MPTP group.



Table.21: Effect of Prosopis chilensis seeds on brain dopamine level in MPTP treated mice.

Groups

g/g brain tissue


Control 3.815 ± 0.055 3.815 ± 0.055 3.815 ± 0.055 3.815 ± 0.055

MPTP 1.257 ± 0.038 *** 1.257 ± 0.038 *** 1.257 ± 0.038 *** 1.257 ± 0.038 ***

100mg/kg PCE 1.376 ± 0.058 *** 1.260 ± 0.018 *** 1.272± 0.053 *** 1.264 ± 0.017 ***

200 mg/kg PCE 1.582 ± 0.041 ***

++

1.268 ± 0.024 ***

1.286 ± 0.043 *** 1.286 ± 0.042 ***

300 mg/kg PCE 1.802 ± 0.040 ***

+++

1.441 ± 0.034 ***

1.758 ± 0.025 ***

+++

1.685 ± 0.012***

+++

300 mg/kg PCE 2.824 ± 0.065 ***

+++

1.948 ± 0.070***

+++

2.794 ± 0.054 ***

+++

2.713 ± 0.089 ***

+++


*** (P<0.001) Vs Control group; ++ (P<0.01),




Table.22: Effect of Prosopis chilensis seed on brain epinephrine level in MPTP treated mice.

Groups

g/g brain tissue


Control 3.288 ± 0.102 3.288 ± 0.102 3.288 ± 0.102 3.288 ± 0.102

MPTP 2.106 ± 0.090 *** 2.106 ± 0.090 *** 2.106 ± 0.090 *** 2.106 ± 0.090 ***

100mg/kg PCE + MPTP 2.210 ± 0.283 ** 2.116 ± 0.062 *** 2.167 ± 0.191 *** 2.190 ± 0.165 ***

200 mg/kg PCE + MPTP 2.252 ± 0.038 *** 2.155 ± 0.094 *** 2.185 ± 0.144 *** 2.241 ±0.115 ***

300 mg/kg PCE +MPTP 2.705 ± 0.059 ***

+++

2.260 ± 0.123 *** 2.606 ± 0.039 ***

+

2.644 ± 0.023 ***

+

300 mg/kg PCE 3.055 ± 0.109 +++ 2.884 ± 0.093 *

+++

3.081 ± 0.059 +++ 3.096 ± 0.035+++

Values are expressed as Mean ± SEM;

* (P<0.05), ** (P< 0.01), *** (P< 0.001) Vs Control group;




Table.23: Effect of Prosopis chilensis seed on brain norepinephrine level in MPTP treated mice.

Groups

g/g brain tissue


Control 3.344 ± 0.054 3.343 ± 0.054 3.343 ± 0.054 3.343 ± 0.054

MPTP 2.148 ± 0.041 *** 2.148 ± 0.041 *** 2.148 ± 0.041 *** 2.148 ± 0.041 ***

100mg/kg PCE + MPTP 2.186 ± 0.048 *** 2.152 ± 0.052 *** 2.151 ± 0.042 *** 2.172 ± 0.122***

200 mg/kg PCE + MPTP 2.254 ± 0.094 *** 2.209 ± 0.065 *** 2.152 ± 0.047 *** 2.250 ± 0.085 ***

300 mg/kg PCE +MPTP 2.636 ± 0.092 ***

+++

2.228 ± 0.075 *** 2.586 ± 0.031 ***

++

2.633 ±0.047 ***

+++

300 mg/kg PCE 3.136 ± 0.078 +++ 2.817 ±0.051 ***

+++

3.061 ± 0.065 **

+++

3.089 ± 0.054+++

Values are expressed as Mean ± SEM (n=6);


++ (P< 0.01), +++ (P<0.001) Vs MPTP group.



Table.24: Effect of Prosopis chilensis seed on brain serotonin level in MPTP treated mice.

Groups

g/g brain tissue


Control 3.407 ± 0.050 3.407 ± 0.050 3.407 ± 0.050 3.407 ± 0.050

MPTP 2.068 ± 0.060 *** 2.068 ± 0.060 *** 2.068 ± 0.060 *** 2.068 ± 0.060 ***

100mg/kg PCE + MPTP 2360 ± 0.038 *** 2.083 ± 0.023 *** 2.198 ± 0.073 *** 2.198 ± 0.066 ***

200 mg/kg PCE + MPTP 2.418 ± 0.051 ***

++

0.193 ± 0.047 *** 2.323 ± 0.090 *** 2.406 ± 0.062 ***

++

300 mg/kg PCE +MPTP 2.571 ± 0.118 ***

+++

2.245 ± 0.067 *** 2.374 ± 0.074 *** 2.489 ± 0.043***

+++

300 mg/kg PCE 3.103 ± 0.039 *

+++

2.790 ± 0.034 ***

+++

3.088 ± 0.055 +++ 3.115 ± 0.047 *

+++



++ (P< 0.01), +++ (P< 0.001) Vs MPTP group.



Table. 25: Effect of Prosopis chilensis seed on lipid peroxidation (nmol/mg protein/hr) level in MPTP treated mice.

Groups

nmol/mg protein/hr

Alcoholic Pet-ether Ethyl acetate Water extract

Control 26.25 ± 3.4 26.25 ± 3.4 26.25 ± 3.4 26.25 ± 3.4

MPTP 67.0 ± 5.6 *** 67.0 ± 5.6 *** 67.0 ± 5.6 *** 67.0 ± 5.6 ***

100mg/kg PCE + MPTP 51.41 ± 1.0 ***

++

61.41 ± 0.9 *** 53.41 ± 1.5 *** 57.16 ± 0.6 ***

200 mg/kg PCE + MPTP 40.5 ± 1.9 **

+++

57.83 ± 1.0 ***

++

47.83 ± 0.7 ***

++

53.20 ± 0.9 ***

+

300 mg/kg PCE +MPTP 29.9 ± 1. +++ 49.79 ± 1.7 +++ 37.08 ± 3.4 +++ 39.62 ± 1.5 *

+++

300 mg/kg PCE 20.8 ± 1. +++ 32.95 ± 1.0 +++ 27.50 ± 1.9 +++ 31.58 ± 1.1 +++



+ (P< 0.05), ++ (P< 0.01), +++ (P< 0.001) Vs MPTP group.



Table.26: Effect of Prosopis chilensis seed on reduced glutathione (µmol/mg protein) level in MPTP treated mice.

Groups

µmol/mg protein


Control 759.90 ± 53.2 759.90 ± 53.2 759.90 ± 53.2 759.90 ± 53.2

MPTP 476.60 ± 24.9 *** 476.60 ± 24.9 *** 476.60 ± 24.9 *** 476.60 ± 24.9 ***

100mg/kg PCE + MPTP 493.77 ± 23.6 *** 483.07 ± 11.7 *** 485.83 ± 26.2 *** 484.32 ± 25.4 ***

200 mg/kg PCE + MPTP 605.50 ± 16.0 **

+

495.37 ± 15.3 *** 573.23 ± 26.9 ** 553.60 ± 22.0 ***

300 mg/kg PCE +MPTP 687.60 ± 10.4 +++ 581.60 ± 9.4 *** 621.97 ± 9.8 *

+

606.70 ± 24.2 **

+

300 mg/kg PCE 734.50 ± 5.3 +++ 700.77 ± 23.0 +++ 726.52 ± 9.4 +++ 730.75 ± 7.7 +++



+ (P< 0.01), +++ (P< 0.001) Vs MPTP group.



Invitro antioxidant studies:

The antioxidant activity of AEPC evaluated by various models.

DPPH:

The AEPC showed the free radical scavenging activity. IC50 value of AEPC

was 320 µg/ml. Results are given in (Table. 27 ; Fig. 111)

Lipid Peroxidation:

Malondialdehyde formation was inhibited by AEPC in rat brain homogenate.

The IC50 value of AEPC was found to be 400 µg/ml (Table. 27 ; Fig. 112).

Nitric Oxide (No) Scavenging:

The AEPC exhibited good free radical scavenging action against NO free

radical. IC50 value of AEPC found to be at 300 µg/ml (Table. 27 ; Fig. 113).

Superoxide dismutase:

AEPC showed the free radical scavenging activity with IC50 value was

found to be 370 µg/ml (Table. 27 ; Fig. 114).



Table. 27 Effect of alcoholic extracts of Prosopis chilensis on different antioxidant

models

S.No.

% inhibition

Conc

µg/ml

DPPH Superoxide

dismutase

Nitric oxide Lipid

Peroxidation

1 3 7.45 7.87 6.83 10.2

2 6 11.89 15.9 11.29 16.9

3 12 21.8 19.18 18.96 21.2

4 25 26.2 23.3 27.33 28.9

5 50 39.5 40.89 39.4 34.7

6 100 50 46.3 47.8 40.1

7 200 57.9 50.25 58.0 46.1

8 400 59 54.7 60.5 51.2

9 800 85.4 82.15 87.9 76.8

10 IC50 (µg/ml) 320 370 300 400




Pharmacognostic evaluation:

Table.28 Phytochemical analysis of PC seed extract showed the presence of

following constituents

Tests Alcoholic Pet. ether Ethyl acetate Water

Flavonoids + - + +

Tannins and Phenolic Compounds + - + +

Steroids - + - -

Carbohydrates + - - +

Alkaloids + - + -

Protein and amino acids + - - +

Saponins + - - +

Fixed oils - + - -

Volatile oils - + - -


Colour : Seeds are light brown

Shape : Kidney shaped, strong

Size : Small


Taste : Characteristic


Microscopic analysis of PC seed powder showed the presence of big ellipsoidal

oil glands, Parenchyma in group, lignified stone cells, round shaped blue coloured

starch grains were present.



Behaviour of PC seed powder with different reagents:


Under visible light under UV light

(365 nm)

Powder as such light brown No Fluorescence

Powder + Picricacid Brownish yellow Light green Fluorescence

Powder + HCl Honey Water green Fluorescence

Powder + H2SO4 Honey Parrot green Fluorescence

Powder + HNO3 Light brown Sky blue Fluorescence

Powder + glacial acetic acid Mud colour green Fluorescence

Powder + FeCl3 Brown dark green Fluorescence

Powder + iodine Reddish brown Cascade green

Fluorescence

Powder + NaOH (aq) Yellowish brown light green Fluorescence

Powder + NaOH (Methanolic) Brownish Water green Fluorescence

Powder + HCl + H2O Light brown No Fluorescence

Powder + H2SO4 + H2O Colourless Sky blue Fluorescence

Powder + HNO3 + H2O Creamish Megenta green

Fluorescence

Determination of Ash value:

Total ash:

Total ash content of PC seed was found to be 4%, where as acid insoluble ash

and water soluble ash was found to be 0.5% and 1% respectively.

Determination of Moisture content:

Moisture content of PC seed powder was found to be 6%.

Prosopis chilensis - Discussion


Discussion:

Alcoholic extract of PC assessed for the neuropharmacological evaluation. PC

extract showed significant increase of spontaneous motor activity, motor coordination

alertness, swimming time in FST, and it also exhibited decreased immobility time in

TST and FST. These findings suggest that Prosopis chilensis seed extract may possess

active principle with central neverous system stimulant activity. Further, PC seed

extract was conformed in FST and TST test. These are antidepressant models. Our

results are similar to earlier reports (Porsolt et al, 1977; Steru et al, 1985; Detke et al,

1995).

Behavious study of MPTP treated mice were carried out in 3 different

parameters. Spontaneous motor activity, exploratory behaviour and motor coordination

were decreased in MPTP treated group, it could be due to motor impairment and

muscle relaxant effect. On treatment with alcoholic extract of Prosopis chilensis

reversed the behavior alterations induced by MPTP on 3rd

day. But maximum effect of

extract was seen on 6th

day of treatment. It might be due to presence of

phytoconstituents like l-dopa polyphenols and flavonoids.

MPTP treated mice not only altered the behavioural response but also reduced

the dopamine in the brain. These evidences suggest that these symptoms are similar to

human Parkinsonism (Mohanasundary et al, 2006). Other amines are also altered in

Parkinson’s disease, Serotonin (Robert et al, 1987), Norepinephrine (Stephen et al,

1984) were much less altered than dopamine.



the body; obviously there is great stress from parkinsons disease. Further, epinephrine







Depression is a common symptom in patients with parkinsons disease.


of serotonin in Parkinson’s disease is also reduced, but this has not bean related to any


metabolite of serotonin, 5-hydroxyindoleacetic acid was lower in depressed than

nondepressed parkinsonian’s. The data suggest that the alterations in serotonin

metabolism in Parkinson’s disease identify a subgroup of patients who prone to

depression (Mayeux et al, 1984).

DA and 5-HT transporters are differentially affected in PD and 5-HT

transporters in the mid brain region may not be affected in relatively early stages of PD

(Sang et al, 2003).

The loss of dopamine and other amines was reversed well by alcoholic extract

of PC because it might be due to cumulative effect of all components present in

alcoholic extract of PC seed. The EAEPC and WEPC had moderate effect it might be

due to presence of specific component extracted in particular solvent. Pet ether extract

of PC did not show significant effect on MPTP treated mice but only plant extract itself

showed activity it could be due to presence of active components in low concentration.

Alcoholic and water extract of the plant might contain L-dopa in low

concentration and polyphenols. Even though if L-dopa present in low concentration its



efficiency was enhanced by some components and adjuvants present in PC seeds

(Hussain et al, 1997).

Oxidative stress leads to organ damage, cell death. In the present study, we

carried out reduced glutathione estimation and assessed lipid peroxidation in brain in

vivo.

Malondialdehyde (MDA) is an end product of lipid peroxidation (Auddy et al,

2003). In the present study, MDA level was increased in MPTP treated mice. This

finding is similar to earlier reports (Amarnath, 2004). Alcoholic and water extract of

PC had good effect on decreased MDA level than other fractions, it could be due to

presence of compounds like phenolic, L-dopa and flavonoids (LU and FOO, 2001).

The reduced glutathione level in MPTP (Mohana sundari et al, 2006) treated

group was observed while their level was restored significantly with alcoholic, water

and ethyl acetate extract of PC. Antioxidant activity was not only due to presence of

flavonoids and polyphenols but also due to presence of alkaloids (Bhattacharya et al,

2010). Preliminary phytochemical analysis of this plant showed the presence of

flavonoids, phenolic compounds amino acids, proteins and alkaloids. Hence, the

antioxidant and free radicals scavenging activity of the plant might be due to presence

of such compounds.

Antioxidant activity of PC was observed in various invitro antioxidant models.

Antioxidants are resistant to oxidative stress by scavenging the free radicals (Youdim

et al, 2001). The AEPC showed marked and dose dependent free radicals scavenging

with Ic50 value of 320 µg/ml, this is similar to earlier reports (Bhattacharya et al, 2010).



The AEPC seed inhibited the lipid peroxidation in a dose dependent manner

with an IC50 value of 400 µg/ml. Hence Prosopis chilensis seed has good antioxidant

property; it could be due to presence of flavonoids (Bhattacharya et al, 2010).

Nitric oxide free radical was inhibited by the alcoholic extract of Prosopis had

active principle with antioxidant property (Marcocci et al, 1994).AEPC seed scavenged

the superoxide free radicals with an Ic50 value of 370 µg/ml, these findings are similar

to earlier reports (Shirwaikar et al, 2004). It could be due to presence of antioxidant

principle present in the PC plant extract.

Macroscopic, microscopic and other Pharmacognostic evaluation may help in

identifying the crude drugs in powder form and also to identify the closely related

species and adulteration. Further, it also used to establish the Pharmacopeial standards.

Conclusion:

Prosopis chilensis improves dopamine loss in brain and also restored the

antioxidants, among all fractions alcoholic and water fractions are more promising for

further studies and may be useful for management of Parkinson’s disease.

Dichrostachys cinerea - Results


DICHROSTACHYS CINEREA:

Results of Pharmacological studies:

Alcoholic extract of DC showed slight sedative effect at a dose of 600 mg/kg or

above during the 24 h period after oral administration. DC extract at a dose of 100,

200, 300 mg/kg produced significant (p<0.01) and dose dependent decrease in

locomotor activity in compared to control group (Fig.115).

Alcoholic extract at given doses (100, 200, 300 mg/kg) showed no significant

(p<0.01) change in reaction time in compared to control group in Eddys Hot Plate test

(Fig.116). Alcoholic extract at a dose of 100, 200, 300 mg/kg did not induce significant

(p<0.01) and dose dependent motor in co-ordination (Fig.117).

The results of forced swimming test exhibited that there was significant

(p<0.01) increase in immobility and significant (p<0.01) decrease in swimming and

climbing behaviour of animals treated at a doses of 100, 200, 300 mg/kg in comparison

to control group (Fig.118, 119, 120).

Results of Tail suspension test (Fig.121) revealed that there was significant

(p<0.01) and dose dependent increase in the immobility time at all dose levels in

treated groups in comparison to control group.

The results of the hole board test are summarized in Fig.122. A significant

(p<0.01) decrease in exploratory behaviour was observed at all dose levels and

followed a dose dependent decrease in comparison to control group.

Invitro antioxidant activity:

An Invitro antioxidant study was carried out in various antioxidant models to

assess scavenging potential of alcoholic extract of Dichrostachys cinerea.



DPPH:

The antioxidant property of alcoholic extract of Dichrostachy cinerea (DC) was

evaluated by DPPH radical scavenging assay.

The antioxidant potential with IC50 value of DC root was found to be 340 µg/ml

(Table.29; Fig.123).

Lipid peroxidation:

The antioxidant property of DC also evaluated by the inhibition of malondialde

hyde formation in rat brain homogenate. The IC50 value of alcoholic extract of DC was

found to be 410 µg/ml (Table.29; Fig.124).

Nitric oxide (No) Scavenging:

The alcoholic extract root of DC root showed promising free radical scavenging

action against NO induced release of free radicals. The free radical scavenging effect of

NO was in a concentration dependent manner. The IC50 values of DC root was found to

be 420µg/ml (Table.29; Fig.125).

Superoxide dismutase:

The alcoholic extract of root of DC exhibited the free radical scavenging action

against highly reactive superoxide to oxygen and hydrogen peroxide at the

concentration of 800 µg/ml showed 86.5 % of O2 inhibition in a concentration

dependent manner. The IC50 value of alcoholic extract of DC was found to be 380

µg/ml (Table.29; Fig.126).



Table. 29 - Effect of alcoholic extracts of Dichrostachys cinerea on different

antioxidant models

S.No.

% inhibition

Conc

µg/ml

DPPH Superoxide

dismutase

Nitric

oxide

Lipid

Peroxidation

1 3 16 12.9 5 12

2 6 24 16.4 14 19

3 12 27 21 18 24

4 25 36 29.7 24 29

5 50 38 32.6 31 35

6 100 42 36 39 38

7 200 50 38.7 43 50

8 400 56 53.7 48 51

9 800 86 86.5 78 72

10 IC50 (µg/ml) 340 380 420 410

Values are mean of 3 replicates)


Results of the preliminary phytochemical analysis carried out on the crude

alcoholic extract indicated the presence of glycosides, steroids, saponins, carbohydrates

and tannins (Table.30).



Table.30 Preliminary phytochemical analysis:

Test Alcoholic Pet. ether Ethyl

acetate

Water

Glycosides + - - -

Carbohydrates + - + +

Alkaloids - - - -

Saponins + - - +

Steroids - + - -

Fats & fixed oils - - - -

Tannins + - + +

Flavonoids + - + +

Macroscopy:

Colour : Outer surface whitish yellow, inner surface white


Taste : Characteristic

Extra feature : Smooth, slightly fractured

Microscopy:

Dichrostachys cinerea powder is yellowish white and free flowing.

Microscopic character of DC powder showed the presence of pitted vessels, tracheid

fibers, cork cells with lignified and brownish matter, polygonal parenchyma and blue

coloured starch grains.



Behaviour of powders with different chemical reagents:


under visible light under UV light

(365 nm)

Powder as such yellowish white No fluorescence

Powder + HCl colour less No fluorescence

Powder + H2SO4 Dull brown Honey colour

Powder + picric acid Yellowish white Parrot green fluorescence

Powder + HNO3 light yellow water green fluorescence

Powder + glacial acetic acid whitish yellow ppt Cascade green

Powder + Ethanol 95% Cream colour Water green

Powder + 5% KOH Mud colour Dark green

Powder + Methanolic (IN) NaOH Ash colour Light green

Powder + HCl + H2O Colourless Sky blue colour fluorescence

Powder + iodine Blakish Parrot green

Powder + NaOH (aq) Colourless Water green

Determination of Total ash:

Total ash content of Dichrostachys cinerea root was found to be 4.2% where as

acid insoluble ash and water soluble ash was found to be 1.1% and 1.8% respectively.


Moisture content of DC root was found to be 3%.

Dichrostachys cinerea - Discussion


Discussion:

The results of the present study indicates that the crude alcoholic extract of the

DC root produced a significant decrease in spontaneous motor activity and alteration of

general behaviour is a good index of CNS depressant activity (Salahdeen and Yemitan,

2006) which could be attributed to the sedative effect of the extract. Rotarod test

revealed a significant loss of muscular coordination and the poor performance in the

tail suspension test, this test is mainly used to screen centrally acting muscle relaxant

(Rakotorina et al, 2001), which could be due to loss of muscular strength. Depressant

drugs increases immobility time in FST and TST, decreases swimming and climbing

behaviour in FST, depending upon the concentration and type of depressant drugs

administered (Poonia et al, 2006). CNS depressant action may be due to presence of

phytochemicals in the crude extract of DC.

The DC root extract possessed CNS depressant activity as indicated by the

significantly reduced alertness, motor coordination, spontaneous motor activity,

climbing and swimming in FST and increased immobility time in tail suspension test

and forced swimming test indicated by CNS depressant effects.

From the present results it may be postulated that DC root extract reduces the

radical to the corresponding hydrazine when it reacts with the hydrogen donors in the

antioxidant principles (Moreno, 2002). The DC root exhibited marked and dose

dependent DPPH radical scavenging activity showing the IC50 value 340µg/ml.

Lipid peroxidation is a complex process whereby polyunsaturated fatty acids of

cellular membranes undergo reaction with reactive oxygen species to yield lipid hydro-

peroxides. The antioxidant activity of the DC root extract was further confirmed by

evaluating the inhibition in production of malondialdehyde (MDA) and related

carbonyl products that are produced as by product of lipid peroxidation induced by Fe2+

ascorbate system in the biomembranes of rat brain homogenate. These carbonyl



products are responsible for DNA damage, carcinogenesis and aging related disease

(Riemersma et al, 2000). The MDA reacts with thiobarbituric acid in specific reaction

medium to produce a strong absorption at 532nm. The DC root extract effectively

inhibited the lipid peroxidation in a dose related manner exhibiting IC50 value of 410

µg/ml. This activity is perhaps related to the H+

ion donating capability of the extract

which scavenges the peroxyl radical to inhibit or terminate peroxide chain (Ohkowa

et al, 1979).

Nitric oxide is a free radical produced in mammalian cells, involved in the

regulation of various physiological processes. However, excess production of NO is

associated with several diseases (Ialenti et al, 1993; Ross, 1993). In the present study

the nitrite produced was reduced by the Alcoholic extract of DC. This may be due to

the antioxidant principles in the extract which compete with oxygen to react with nitric

oxide (Marcocci et al, 1994) there by inhibiting the generation of nitrite.

SOD is the one of the important free radical scavenging enzyme present in the

body. The radical scavenging activity of SOD is effective only when it is followed by

the actions of catalase. SOD generates hydrogen peroxide as a metabolite which is

more toxic than oxygen radicals and requires to be scavenged by catalase (Harman,

1991). In the present study alcoholic extract of DC root showed antioxidant scavenging

activity by inhibiting the hydrogen peroxide metabolite. It might be due to presence of

antioxidant compounds like flavonoids. Pharmacognostic evaluation of DC root was

carried out in order to identify the correct identification of this plant and to differentiate

the closely related other species of Dicrostachys cinerea these parameter may be

helpful in identification of the plant.



Conclusion:

Present investigation confirms significant CNS depressant and invitro

antioxidant activity of alcoholic extract of DC further studies are need to confirm the

identity of bioactive principles responsible for these actions by the root of

Dichrostachys cinerea.

Capparis zeylanica - Results


CAPPARIS ZEYLANICA:

Acute toxicity studies and gross behavioural changes:

Alcoholic extract of root bark of Capparis zeylanica was tested upto 3200

mg/kg the extract showed slight behaviour change like sedation at a dose of 800 mg/kg

and above.

Assessment of Neuropharmacological activity:

The animals treated with 100, 200, 300 mg/kg showed significant (P<0.01) and

dose dependent decrease in locomotor activity in comparison to control group

(Fig.127). Alcoholic extract at a dose of 100, 200 mg/kg showed no significant change

in reaction time in comparison to control group (Fig.128) in Eddys hot plate test.

At given doses significant and dose dependent decrease was observed in the

motor co-ordination (Fig.129).

The results of forced swimming test revealed that there was significant increase

(P<0.01) in immobility and significant decrease (P<0.01) in swimming and climbing

behaviour of animals at 100, 200, 300 mg/kg in comparison to control group (Fig. 130,

131,132).

Alcoholic extract exhibited significant and dose dependent increase in the

immobility time at all dose levels compared to control group in tail suspension test

(Fig. 133).

The results of the hole board test are summarized in fig.134. A significant

decrease in the number of head dipping was observed at all dose levels tested and

followed a dose dependent decrease in comparison to control group.



Invitro antioxidant studies:

Invitro antioxidant studies of alcoholic extract of Capparis zeylanica root bark

was carried out in various antioxidant models to asses scavenging potential.

Different concentrations ranging from 3-800µg/ml of the alcoholic extract of

Capparis zeylanica were tested for its antioxidant activity. It was observed that free

radicals were scavenged by the test compounds in a concentration dependent manner.

The maximum percentage inhibition in all the models like DPPH, nitric oxide, lipid

peroxidation and superoxide dismutase with IC50 value of 340 µg/ml in lipid

peroxidation and with an IC50 value of 430 µg/ml in Nitric oxide. The activity was

moderate in the remaining antioxidant models (Table.31, Fig.135, 136, 137 and 138).

Table. 31: Effect of alcoholic extract of Capparis zeylanica on different

antioxidant models

S.No.

% inhibition

Conc

µg/ml

DPPH Superoxide

dismutase

Nitric

oxide

Lipid

Peroxidation

1 3 9.6 8.78 8.2 15.2

2 6 18.4 16.42 11.43 19.3

3 12 20.3 19.7 15.5 25.4

4 25 25.3 24.06 19.3 28.5

5 50 31.2 29 25.3 42.3

6 100 38.4 34.5 30.3 45

7 200 45.6 40.6 37.3 53

8 400 54 49.6 49 58.3

9 800 79.4 78.5 84 82

10 IC50 (µg/ml) 410 400 430 340





Table.32: Preliminary phytochemical analysis of alcoholic extract of Capparis

zeylanica root bark showed the presence of following constituents.

Test Alcoholic Pet. ether Ethyl acetate Water

Glycosides + - - +

Carbohydrates + - - +

Alkaloids ≤ - ≤ -

Tannins - - - -

Flavonoids + - + +

Steroids - + - -

Saponins + - - +

Fats & Fixed oils - - - -

+ = Positive, - = Negative, ≤ = Faint positive

Macroscopy:

Colour : Outer surface of root bark brown, mild rough, inner surface of root bark

light yellow, smooth

Taste : Slightly bitter


Microscopy:

Microscopic analysis of CZ root bark powder showed the presence of lignified

xylem fibers, stone cells. It also contains medullary rays, polygonal cork cells with

brown content, numerous compound and simple starch grains, calcium oxalate crystals

and oil globules.



Behaviour of powder with different chemical reagents:

Reaction Visible light UV light

(365 nm)

Powder as such light brown No fluorescence

Powder + HCl brown Mud colour fluorescence

Powder + H2SO4 honey colour sky blue colour fluorescence

Powder + picric acid brownish yellow blue fluorescence

Powder + HNO3 mud colour water green fluorescence

Powder + glacial acetic acid light brown cascade green fluorescence

Powder + Ethanol 95% brown colour parrot green fluorescence

Powder + 5% KOH light yellow dark green fluorescence

Powder + Methonolic (IN) NaOH honey colour light green fluorescence

Powder + HCl + H2O light green leaf green fluorescence

Powder + NaOH (aq) blakish blue olive green fluorescence

Powder + iodine yellowish brown honey colour fluorescence

Quantitative Pharmacognostic Evaluation:

Determination of Ash Content:

Total Ash:

The total ash after incineration of root bark of capparis zeylanica powder was

found to be 5.4% where as acid insoluble ash and water soluble ash was found to be 2%

and 3% respectively.


Moisture content of Capparis zeylanica root bark powder was found to be 3.8%.

Capparis zeylanica - Discussion


Discussion:

In this study, alcoholic extract of (CZ) root bark was investigated for its effect

on the central nervous system. The plant extract possessed CNS depressant activity as

indicated by the significantly reduced alertness, motor co-ordination, spontaneous

motor activity, climbing and swimming in FST and increased immobility time in tail

suspension test and forced swimming test.

Decreased spontaneous motor activity could be attributed to the sedative effect

of the extract (Rakotonirina et al, 2001). The extract also reduced the time spent on the

rotarod, a test mainly used to screen centrally acting muscle relaxants (Rakotonirina

et al, 2001). The (CZ) extract may have muscle relaxant activity, which could be due

to CNS depressant activity. Hole board test evaluates the effect of extract on alertness.

Immobility period in FST and TST reflects a state of despair that can be reduced by

several agents, which are therapeutically effective in human depression. Extract of CZ

increased the immobility in FST and TST indicating CNS depressant effects.

The root is used as analgesic but in hot plate test reaction time was not altered

indicating that opioid mechanisms are not involved in the analgesic effect of CZ. The

alcoholic extract may have active constituents with CNS depressant activity.

Result of the present study, the alcoholic extract of root bark of Capparis

zeylanica showed the antioxidant activity against various invitro antioxidant models.

Root bark of CZ showed marked and dose dependent free radical scavenging activity in

DPPH radical scavenging assay showing the IC50 value of 410 µg/ml.

The malondialdehyde formation was inhibited by the alcoholic extract of CZ

root bark in a dose dependent manner. CZ root bark extract had an H+

ion donating

capacity which scavenges the peroxyl radical (Ohkowa et al,, 1979).

Capparis zeylanica - Discussion


The Alcoholic extract of root bark of CZ inhibited the excess nitric oxide

produced by the incubation of solutions of sodium nitroprusside in standard phosphate

buffer at 250C. This activity might be due to the presence of phytoconstituents having

antioxidant property.

CZ root bark extract catalyses the dismutation of the highly reactive superoxide

to oxygen and hydrogen peroxide (Kamalakkannan et al, 2003). The IC50 value of this

root bark extract was 400 µg/ml.

Pharmacognostic evaluation of the root bark of Capparis zeylanica. It provides

diagnostic characters useful for the identification of the closely related drugs.

Conclusion:

The preliminary pharmacological studies on the alcoholic extract of Capparis

zeylanica root bark indicate that the root bark has active principles with CNS

depressant activity. However, further pharmacological investigations are required to

understand its underlying mode of action on the CNS.

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