a review on amorphophallus paeoniifolius - rspublication

INTERNATIONAL JOURNAL OF ADVANCED SCIENTIFIC RESEARCH AND TECHNOLOGY

ISSUE 2, VOLUME 2 (APRIL 2012) ISSN: 2249-9954

A REVIEW ON AMORPHOPHALLUS PAEONIIFOLIUS

P.Madhurima#1

, I. J. Kuppast#2

, K. L. Mankani#3

,

DEPARTMENT OF PHARMACOLOGY, NATIONAL COLLEGE OF PHARMACY,

BALRAJ URS ROAD, SHIMOGA, KARNATAKA, INDIA.

#1, Phone no: 9686232304, [email protected]



ABSTRACT:

Herbs have been used by people for longer than we have been keeping written record. Originally

they were found in the wild, by the gatherers and used for a lot of different things. They were used to

flavour food, as a source of nutrition, as medicines1. Amorphophallus paeoniifolius known as

Elephant foot yam is basically a crop of south East Asian origin. In India, it is commonly known as

"Suran" or "Jimmikand". It grows in wild form in Philippines, Malaysia, Indonesia and other South

East Asian countries. This tuber is consumed by many people as a food and widely used in many

Ayurvedic preparations. In recent years the popularity of complementary medicine has increased.

Over 50% of all modern drugs are natural product origin and they play an important role in drug

development programs of the pharmaceutical industry. Epidemiological evidence suggests that

dietary factors play an important role in human health and in the treatment of certain chronic diseases

including cancer. The tuber is reported to have antiprotease activity, CNS depressant activity,

analgesic activity, and cytotoxic activity2.

Key words: Amorphophallus paeoniifolius, antioxidant activity, antitumor activity, analgesic

activity, antidiarrhoeal activity, CNS depressant activity, phytochemical constituents, cytotoxicity.

Corresponding Author: P. Madhurima

INTRODUCTION:

Herbal drugs occupy a special place in the world of pharmaceuticals. Side-effects of

conventional medicines, efficiency of plant-derived drugs and growing interest in natural products

have increased interest in medicinal plants3. Herbal remedies have been developed through

knowledge of herbs, which is a ray of hope for several patients. Amorphophallus belongs to the

family Araceae4.

ORIGIN AND DISTRIBUTION:

It is a crop of South East Asian origin. It grows wild form in Philippines, Malaysia, Indonesia

and other South East Asian countries. None of them are found in Americas although a remarkably

similar but not closely related genus, Dracontium, has evolved there. Most species are endemic. They

grow preferentially on distributed grounds, such as secondary forests5.

DESCRIPTION:

It is a stout cormous herb, corm depressed globose, 25-30cm across.

Stem:

It is a tuber depressed-globose, dark brown, root scars prominent, annulate, offlets produced

every season, thick and rhizomatous.

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Leaf:

It is solitary or two; petiole to ca. 2 meters long and 20cm in diameter, background color pale to

dark green or blackish green. Lamina is highly dissected, to about 3 meters in diameter, leaflets

rounded, oval, ovate, obovate, elliptic, elliptic-oblong, elliptic-lanceolate or lanceolate, acuminate, 3-

35 cm long, 2-12 cm in diameter, upper surface midgreen, lower surface midgreen to pale green.

Flowers:

In flowers inflorescence is short-peduncled, peduncle is 3 to 20 cm long, about 1 to 8 cm in

diameter, usually paler and smoother than petiole; spathe campanulate, broader than long, 10 to

about 40 cm long, 15 to about 60 cm in diameter, base and limb often separated by a shallow

constriction, limb spreading, strongly undulate, base outside very variable, background color ranging

from pale green to dark brown, usually with large and small, circular paler spots, base inside lower

part deep maroon, upper zone dirty whitish or pale pinkish, limb outside as base but with more

prominent maroon flushes, especially near the margin, limb inside usually glossy dark maroon, base

within densely verrucate, verrucae variable, mostly conical, fleshy. Fruits are produced in spikes.

Individual fruits are ellipsoid to globular, orange to red.

Seeds:

Seeds are almost as large as fruits6.

Habitats:

Loose and leafy detritus are found in moist shady habitats. Tropical conditions are seen in

secondary forests, shrub forests and grasslands in arid valley areas and at elevations below 750

metres.

KNOWN HAZARDS:

Although no specific mention has been seen for this species, it belongs to a family where most

of the members contain calcium oxalate crystals. This substance is toxic fresh and, if eaten, makes

the mouth, tongue and throat feel as if hundreds of small needles are digging in to them. However,

calcium oxalate is easily broken down either by thoroughly cooking the plant or by fully drying it

and, in either of these states, it is safe to eat the plant. People with a tendency to rheumatism,

arthritis, gout, kidney stones and hyperacidity should take especial caution if including this plant in

their diet7.

PHYTOCHEMICALS:

The qualitative chemical tests for various phytoconstituents were carried out for methanolic and

petroleum ether extracts of Amorphophallus paeoniifolius. The phytochemical tests were performed

for the presence of Alkaloids (Mayer’s test, Dragendorff’s test, Wagner's test), Glycosides

(Raymond’s test, Legal’s test), specific glycosides like Saponin Glycosides (Froth Test, Hemolysis

test), Anthraquinone Glycosides (Borntrager's test, Modified Borntrager's test), Cardiac Glycosides

(Kedde’s test, Keller killiani test), Cyanogenetic Glycosides, tannins and Phenolic Compounds

(Gelatin test, Ferric chloride test, Alkaline reagent test), Flavonoids (Shinoda test, Zinc

Hydrochloride reduction test, Alkaline reagent test), Proteins and Amino Acids (Millons test,

Ninhydrin test), Sterols & Triterpenoids (Libermann-Buchard test, Salkowski test), Carbohydrates

(Molisch's test, Benedict's test, Barfoed's test, Camnelisation, Selwinoff’s test, Fehling's test), Fats &

Fixed Oils (Stain test, Saponification test).

Phytochemical screening of methanolic extract indicated the presence of steroids, flavonoids,

alkaloids and carbohydrates. Phytochemical screening of Petroleum ether extract indicates the

presence of steroids, fats and fixed oil. The methanolic extract was found to be reddish in colour

having characteristic odour viscous consistency while the petroleum ether extract was found to be

pale yellow in color having viscous consistency8.



TRADITIONAL USES:

The root is carminative, restorative, stomachic and tonic. It is dried and used in the treatment

of piles and dysentery. The fresh root acts as an acrid stimulant and expectorant, it is much used in

India in the treatment of acute rheumatism.

The tubers are anodyne, antiinflammatory, antihaemorrhoidal, haemostatic, expectorant,

carminative, digestive, appetizer, stomachic, anthelmintic, liver tonic, aphrodisiac, emmenagogue,

rejuvenating and tonic. They are traditionally used in arthralgia, elephantiasis, tumors,

inflammations, hemorrhoids, hemorrhages, vomiting, cough, bronchitis, asthma, anorexia, dyspepsia,

flatulence, colic, constipation,helminthiasis hepatopathy, spleenopathy, amenorrhea, dysmenorrhoea,

seminal weakness, fatigue, anemia and general debility. The tuber is reported to have antiprotease,

analgesic, cytotoxic, anti-inflammatory and CNS depressants activities7.

ANTI-TUMOUR AND ANTIOXIDANT ACTIVITIES:

Cancer is a class of diseases in which a group of cells display uncontrolled growth (division

beyond the normal limits), invasion (intrusion on and destruction of adjacent tissues), and sometimes

metastasis (spread to other locations in the body via lymph or blood). These three malignant

properties of cancers differentiate them from benign tumors, which are self-limited, and do not

invade or metastasis.

In-vivo anti-oxidant activity:

DPPH (Diphenylpicrylhydrazyl) radical scavenging activity:

The scavenging activity of Amorphophallus Paeonifolius extract was measured in terms of

hydrogen donating or radical scavenging ability using the stable radical DPPH Blois et al.,

(1958).[10] 0.1Mm solution of DPPH in ethanol was prepared and 1.0ml of this solution was added

to 3 mlof extract solution and standard in water at different concentrations (10-100μg/ml). 30min

later absorbance was measured at 517nm. Lower absorbance of the reaction mixture indicated higher

free radical scavenging activity. The capability to scavenge the DPPH radical was calculated using

the following equation.

Reducing power:

The reductive potential of the extract was determined according to the method of Oyaizu et

al., (1986).[11] The different concentration of extracts and standard in 1ml of distilled water was

mixed with phosphate buffer (2.5ml, 0.2m, pH6.6) and potassium ferricyanide [K3Fe(CN)6]

(2.5ml,1% w/v). The mixture was incubated at 50oC for 20min.2.5ml of trichloroaceticacid

(10%w/v) was added to the mixture, which was then centrifuged for 10min at 1000rpm. The 2.5 ml

of supernatant was mixed with distilled water (2.5ml) and Fecl3 (0.5ml, 0.1% w/v), and mixed.

Absorbance was measured at 700nm in a spectrophotometer. Higher absorbance of the reaction

mixture indicated greater reductive potential.

Superoxide anion scavenging activity assay:

The scavenging activity of Amorphophallus Paeonifolius towards superoxide anion radicals

was measured by the method of Ni-shimiki et al.,(1972).[12] and slightly modified. About 1ml of

nitroblue tetrazolium solution (156μm in 100Mm phosphate buffer, pH 7.4) and 0.1ml of different

concentrations of extract and standard in water were mixed. The reaction was initiated by adding

% inhibition =Aconc – Atest

Aconc × 100



100μl of phenazinemethosulphate(PMS) solution (60μM) in 100Mm phosphate buffer(pH7.4) to the

mixture. The reaction mixture was incubated at room temperature for 5 min and the absorbance at

560nm was measured against reagent blank in spectrophotometer. (Quercetin was used as standard).

The cancer was inducted by using chemical induction method using 7, 12-Dimethyl Benz (A)

Anthracene(DMBA) DMBA was purchased from sigma chemicals, Mumbai, India. 120 mg DMBA

was dissolved in 24 ml olive oil. It forms a yellow colour solution. DMBA solution was given to

animals orally with the help of oral feeding needle at a dose of 25 mg/kg.

Table1. Estimation of Hematological and Bio-Chemical Parameters

Group

(n=6)

Control

DMBA

Only

DMBA + DMBA +

AP-Tuber

extract

(50mg/kg)

DMBA+AP

Tuber extract

(100mg/kg)

DMBA +

Tamoxifen (10mg/kg)

WBC (x109/L)

6.95±0.13

14.09±0.14

a 7.10±0.12

a 7.08±0.22

a 7.21±0.40

a

RBC(x1012

/L)

13.26±0.36

10.13±0.71 a

12.43±0.27

a

13.43±0.75 a

12.76±0.10a

Hb(g/dL)

13.26±0.36

10.13±0.71 a

12.5±0.44 b

13.43±0.75 a

12.76±0.17a

NEUTROPHIL

(%)

8.96±0.18

17.13±0.18 a

9.26±0.31 a

9.03±0.54b

8.9±0.17 a

SGPT

78.66±2.251

112.33±28.7

8 a

87.66±9.89

3c

74±7.797 a

88.66±3.141ns

SGOT

131±22.68

225.66±27.0

7 a

140.33±18.

99 a

148.66±18.64 a

154.66±7.607a

Creatinine

0.633±0.051

0.733±0.051

a

0.733±0.05

1 a

0.7±0.001 a

0.666±0.051 a

Urea 38.56±0.459

47.5±4.588 a

30.93±4.51

9 a

32±2.968a

32.26±5.643 a

a-P< 0.001, b-P<0.01,c-P<0.05, ns-non significant

Enzymatic Anti-oxidant activity:

Estimation of catalase activity:

Catalase activity was measured by method described by Sinha et.al.(1972). The

reaction mixture (1.5ml, vol) contained 1.0ml of 0.01M phosphate buffer (pH7.0), 0.1ml of

tissue homogenate and 0.4ml of 2M H2O2.The reaction was stopped by the addition of 2.0ml

of dichromate-acetic acid reagent (5% pot. dichromate and glacial acetic acid were mixed in

1:3 ratio). Then the absorbance was read at 620nm; CAT activity was expressed as μMol of

H2O2 consumed/min/mg protein.

All the experimental animals were killed by cervical decapitation after the

experimental period. For the estimation of non-enzymic and enzymic antioxidants, tissue

(liver and kidney) was minced and homogenized (10% w/v) in 0.1 M phosphate buffer (Ph

7.0) and centrifuged for 10 min and the resulting supernatant was used for enzyme assays.



Estimation of Superoxide dismutase (SOD) Activity:

The activity of superoxide dismutase was assayed by the method of Kakkar

et.al..(1984). Breifly in a test tube; 0.5ml of supernatant tissue homogenate was taken. To this

1.5ml of carbonate buffer(pH10.2),0.5ml of 0.1Mm EDTA and 0.4ml of epinephrine was

added and the OD was taken at 480nm.Epinephrine was added just before taking the OD.

This activity is to be expressed as units/min/mg protein.

Estimation of Glutathione peroxidase (GPx) Activity:

GPx activity was measured by method described by Rotruck et.al.(1973). Briefly, the

reaction mixture contained 0.2ml 0.4M Phosphate buffer (pH7), 0.1ml 10Mm sodium azide,

0.2ml tissue homogenized in 0.4M Phosphate buffer(pH7), 0.2ml glutathione, and 0.1ml

0.2Mm H2O2. The contents were incubated for 10min at 37o C, 0.4ml 10% TCA was added

to stop the reaction and centrifuged at 3200 rpm for 20min.The supernatant was assayed for

glutathione content using Ellman’s reagent(19.5g 5,5’-dithiobisnitrobenzoic acid (DTNB) in

100ml 0.1% sodium citrate). The activity is to be expressed as μMol of GSH

consumed/min/mg protein.

Non-enzymatic anti-oxidant activity:

Estimation of reduced glutathione (GSH) activity:

GSH was determined by the method of Ellman et al., (1959). [17] A known weight of

tissue homogenized in phosphate buffer from this 0.5ml was pipetted out and precipitated

with 2ml of 5% TCA 1ml of the supernatant was taken after centrifugation at 3200 rpm for

20min and added to it 0.5ml of Ellman’s reagent and 3ml of phosphate buffer (pH 8.0). Then

the absorbance was read at 412nm. The values were expressed as mg/100 g tissue.

Anti –tumor activity:

Experimental design for treatment oriented study:

Experimental rats were divided into 5 groups of six animals each and received the

following treatment for 90 days. Group I- Control rats given only Saline (p.o.), Group II-

Rats given DMBA only 25 mg/kg (p.o.), Group III- Rats treated with DMBA + AP tuber

extract 50mg/kg (p.o.), Group IVRats treated with DMBA + AP tuber extract 100mg/kg

(p.o.), Group V- Rats treated with DMBA + TAM 10 mg/kg (p.o.)

Hematological studies:

In this 90 days study both group of animals were treated with respective extracts and

standard drugs via oral route. Body weight was taken every week till 17th week. After the

completion of treatment, blood was collected for hematological parameters estimation like

RBC, WBC (Total and differential count by Improved Neubauer’s counting chamber method

using Haeyem’s fluid) Hb (by sahli’s Haemoglobinometer method) and Neutrophil by

Improved Neubauer’s counting chamber method.

Enzymatic antioxidant activity



Table2. Antioxidant activity of Amorphophallus paeoniifolius

Bio chemical parameters:

Aliquot of blood was collected and centrifuged at 5000 rpm for 5 min to separate the

serum. This serum was used for estimation of SGOT by Optimized UV- test according to

Group

(n=6)

Control DMBA

only

DMBA +

AP-Tuber

extract

(50mg/kg)

DMBA + AP-

Tuber extract

(100mg/kg)

DMBA +

Tamoxifen

(10mg/kg)

Estimation of

Catalase

Activity

Liver 317.4±70 287.9±10a 443.7±27.8

a

602.5±21.21 a 483.3±29.63

a

kidney 279.13±37.

55

189.4±38.1

1a

540.78±30.8

a

717.69±33.9 a 827.02±22.88

a

Estimation

of

Superoxide

Dismutase

(SOD)

Activity

Liver 40.84±8.56 4.22±2.31a 31.63±3.35

a

53.42±4.02 a 63.82±7.17

a

kidney 36.6±5.92 16.35±1.85a 30.29±3.36

b 24.22±9.20

ns 73.41±1.28

a

Estimation

of

Glutathione

Peroxidise

(GPx)

activity

Liver 206.04±28.

88

97.41±11.7a 132.64±43.8

9 a

290.66±17.09

a

230.94±8.27 a

kidney 269.71±11.

4

87.37±13.1

2a

227.23±39.9

8 a

260.45±36.80

a

291.59±6.00 a

Non-enzymatic antioxidant activity

Estimation

of reduced

glutathione

(GSH)

Liver 851.47±10

5.2

358.36±59.

4a

907.66±200.9c

1034.03±44.52

a

1015.20±166.8 a

kidney 837.95±94

.7

400.74±68.

84a

1041.61±55.6

9 a

1136.86±93.07

a

1161.12±112.4 a



IFCC (International Federation of Clinical Chemistry and Laboratory Medicine), SGPT by

Kinetic UV test, according to the International Federation of Clinical Chemistry and

Laboratory Medicine, serum creatinine by Jaffe Method (modified), serum urea by Urease-

GLDH enzymatic UV test method .

Estimation of protein:

Procedure described by Lowery et.al. (1951) was used for protein estimation. The

method was based on the biuret reaction, formation of a protein-copper complex and

reduction of phosphomolybdo tungstate reagent (Folin-ciocalteu phenol reagent) by tyrosine

and tryptophan residues of protein to form a coloured product.

Statistical Analysis:

The data for various parameters were analyzed using analysis of variance (ANOVA)

Tukey, followed by compared of all pairs of column.

The ethanolic extract of Amorphophallus Paeonifolius has shown significant

antitumor and antioxidant effect in animals. This may be due to presence of flavanoids in

ethanolic extract of Amorphophallus Paeonifolius. The present preliminary investigation

suggests that Amorphophallus Paeonifolius tuber stimulates both cellular and humoral

immunity. Further studies have elucidate the exact antitumor mechanism of Amorphophallus

Paeonifolius tuber9.

ANTI-DIARRHOEAL ACTIVITY:

Acute diarrhoea is one of the principal causes of death in the infants, particularly in

developing countries. An array of medicinal plants with anti-diarrhoeal properties has been

widely used by the traditional practitioners; however, the efficacy of many of these anti

diarrhoeal conventional medicines has not been scientifically evaluated. Hence the present

study was under taken to explore anti-diarrhoeal activity of Amorphophallus paeoniifolius

leaves.

Swiss Albino rats of either sex weighing 150-180 g were used for castor oil-induced

anti-diarrhoeal activity. The animals were kept at 27 ± 2°C, relative humidity 55-65% and

light and dark cycles of 12 and 12 h, respectively, for 1 week before and during the

experiments. All animals were fed standard animal feed and water ad libitum before the

experiments. All the experiments were performed in the morning for the care of the

laboratory animals.

The diarrhoea was clinically apparent in all the animals of control group, (for the next

4 hour) 30 min after administration of castor oil. This was markedly reduced (88% inhibition)

by the oral administration of Loperamide (positive control), 3 mg/kg. Again remarkable

reduction in the number of defaecation as well as diarrhoeic faeces over 4 hours was achieved

in treated group with ExAP, which was found to be dose dependent. The 100 mg/kg, i.p. dose

of the extract showed 46% percentage inhibition in defecation. The percentage inhibition in

defecation was 56% and 69% at the dose of 200 and 400mg/kg respectively, which was

statistically significant (p<0.05).



Table3. Anti-diarrhoeal activity:

Values are Mean±SEM, n=6; * p<0.05 Significant Vs Control

Among the different doses, 200 and 400mg/kg exhibited prominent antidiarrhoeal

activity. Both the doses reduced the total number of faeces as well as total number of wet

faeces in test animals. But the effect was prominent at 400mg/kg, i.p. dose of extract.

Although the investigated plant may be useful in a wide range of diarrhoeal states; further

studies are needed to completely understand the mechanism of anti-diarrhoeal action of A.

paeoniifolius leaves10

.

SYNERGISTIC DEPRESSANT ACTIVITY:

Pharmacodynamic synergism results from two drugs directed at a similar receptor

target or physiological system. Synergistic activity of tea samples with antibiotics showed

best response against the bacteria. The synergistic effect from the association of antibiotic

with plant extracts against resistant bacteria enables the use of the respective antibiotic when

it is no longer effective by itself during therapeutic treatment. Generally neurotransmitters are

involved in the work of regulation of central nervous system (CNS) activity. GABA A

receptors are located post-synaptically and they mediate postsynaptic inhibition by increasing

the Cl- permeability and hyperpolarizing the cell. GABAA receptors are target for the several

important centrally acting drugs like benzodiazepines, barbiturates, etc. Similarly the tuber of

the Ayurvedic plant A. paeoniifolius were found to have CNS depressant activity in mice. So

the study was planned to find out the interaction of the extracts with the benzodiazepines and

barbiturates receptor agonist.

Male Swiss albino mice (20-25 g) were obtained from animal house. The animals

were housed under CPCSEA specified environmental condition and fed with standard diet,

water ad libitum.

The CNS activity was evaluated using Actophotometer and rota-rod apparatus. The

petroleum ether extract of A. paeoniifolius was administered in the form of suspension in 5%

v/v Tween 80 as vehicle. The standard drug diazepam and phenobarbitone was administered

in the form of suspension in 5% v/v Tween 80 as vehicle. The male animals were divided into

twelve groups each composed of six animals as follows.

Faecal output

Groups Dose

Total no of

faecal output

during 4

hour

Total no of

wet faeces

during 4 hour

% Inhibition

Group 1

(Saline)

10 ml/kg, i.p. 13.6± 2.4 10.8±1.8 -

Group 2

(Loperamide)

3 mg/kg, orally 1.8±0.43* 1.2±0.41* 88

Group 3 (EAP) 100 mg/kg, i.p. 7.4± 0.818* 5.8± 0.47* 46

Group 4 (EAP) 200 mg/kg, i.p. 6.2± 0.93* 4.8±0.53* 56

Group 5 (EAP) 400 mg/kg, i.p. 4.8±0.62* 3.4±0.77* 69



Group I: Control (received 5% Tween 80 at the dose of 10 ml/kg, i.p.)

Group II: PEAP (100 mg/kg, i.p.)

Group III: PEAP (300 mg/kg, i.p.)

Group IV: PEAP (1000 mg/kg, i.p.)

Group V: Diazepam (0.1 mg/kg, i.p.)

Group VI: Diazepam (0.5 mg/kg, i.p.)

Group VII: Diazepam (1 mg/kg, i.p.)

Group VIII: Phenobarbitone (1 mg/kg, i.p.)

Group IX: Phenobarbitone (5 mg/kg, i.p.)

Group X: Phenobarbitone (20 mg/kg, i.p.)

Group XI: PEAP (250 mg/kg) and diazepam (0.5 mg/kg, i.p.)

Group XII: PEAP (250 mg/kg) and phenobarbitone (12 mg/kg, i.p.)

After 1hr of the administration of the drug, the animals were placed in an

actophotometer for 10 min and the locomotor activity was observed.

From the dose response curve of each drug, the effective doses were calculated. The

synergistic activity of petroleum ether extract in combination with either diazepam (Group

XI) or phenobarbitone ((Group XII) was evaluated.

The petroleum ether extract of A. paeoniifolius was found to have CNS depressant

activity. Phenobarbitone and diazepam are well known CNS depressant drugs and also

showed CNS depressant activity in a dose-dependant manner. A significant synergistic effect

of the petroleum ether extract with diazepam was found whereas there was little synergistic

effect with phenobarbitone. Both of the phenobarbitone and diazepam exert their CNS

depressant effect by acting on the GABA A receptor. The GABA A receptor has 5 subunits (α,

2ί, γ, δ). The α subunit has the benzodiazepine-binding sites and the barbiturate-binding sites

are located on the ί subunit. The drugs phenobarbitone and diazepam bind with their

respective binding sites and give their effect. From the results, it was concluded that pet-ether

extract has more synergistic activity of the CNS depression with diazepam than

phenobarbitone. As the petroleum ether extract showed synergistic effect with diazepam but

not with the phenobarbitone, the components present in the petroleum ether extract may bind

with the α subunit and facilitate the GABA mediated Cl- channel opening, thus

hyperpolarizes the cell and show CNS depressant action. Diazepam is a benzodiazepine

receptor agonist. So the extract has agonistic activity with benzodiazepine receptor, which

might similar to that of diazepam. Further investigations are needed for better understanding

of molecular mechanism of action and signal transduction of the components present in

petroleum ether extract of A. paeoniifolius regarding CNS depressant activity11

.

8



ANALGESIC ACTIVITY:

The methanol extract of the tuber of Amorphophallus paeoniifolius was studied for its

in-vivo analgesic activity by using the tail-flick method and the acetic acid induced writhing

response in mice. The safety of extract in animals was confirmed from the acute toxicity

studies. The time course study was performed to find the peak time for the maximum

analgesic activity. The effective dose of the extract for analgesic activity was calculated from

dose-response curve by using the tail flick and acetic acid induced writhing method. In both

of the cases diclofenac sodium was used as standard. For the assessment of analgesic activity

in each method the animals of male sex were divided into five groups each composed of six

animals. All groups received intraperitoneal injection (maximum 1 ml as per ethical norms).

Group I: Control animals received 5% Tween80 at the dose of 10 ml/kg.

Group II: Animals received methanol extract at the dose of 250 mg/kg.

Group III: Animals received methanol extract at the dose of 500 mg/kg.

Group IV: Animals received standard Diclofenac sodium at the dose of 5 mg/kg.

Group V: Animals received standard Diclofenac sodium at the dose of 10 mg/kg.

Table 4. Analgesic activity:

n=6 Values are Mean±SD in each.

Comparison were made between group I Vs II, III, IV, V and values *p>0.05, **p<0.01.

In both of tail flick method and acetic acid induced writhing response method; the

intraperitoneal administration of methanol extract of Amorphophallus paeoniifolius tubers

(250, 500mg/kg) induced a significant analgesic activity in a dose-dependent manner. The

plant may have the phytoconstituents which inhibit cyclooxygenase enzyme or act on central

opioid receptors (&receptors)12

.

CYTOTOXIC ACTIVITY:

Amorphophallus paeoniifolius, belongs to the family Araceae, widely distributed in

tropical and sub-tropical regions and are extensively used in South India for various diseases.

The study evaluates the cytotoxic property of different solvent extracts of Amorphophallus

paeoniifolius tuber using Allium cepa L. root tip cells and HEp-2 cell lines as two model

Group Flicking response of tail

%increase in

analgesic

activity

Before treatment After treatment Difference

I 0.85±0.25 0.96±0.77 0.11 1.10

II 1.06±2.39 6.87±2.11 5.81 58.10

III 1.21±0.72 9.1±1.16 7.89 78.90

IV 1.13±0.3411 7.66±0.98 6.53 65.30

V 1.2±0.58 9.47±0.88 8.27 82.70



invitro systems. Of the seven different extracts of Amorphophallus tuber were tested, the

mitotic index and cytolytic index were found to be high in petroleum ether and ethanol

fractions when compared with other solvent extracts. The magnitude of cytotoxicity was

predominant in petroleum ether extract and ethanol extract and displayed a antiproliferative

activity on Hep2 cells. Thus the present study confirms the cytotoxic property of

Amorphophallus paeoniifolius and also demonstrated the role of Amorphophallus

paeoniifolius used in the traditional medicine13

.

EFFECTS ON CENTRAL NERVOUS SYSTEM IN MICE:

A Central Nervous System activity refers to physiological depression of the central

nervous system, general or local anesthesia, relaxation of skeletal muscles, or anticonvulsant

activities. Many depressants and anesthetics acting on the central nervous system do so by

increasing the activity of a particular neurotransmitter known as gamma-aminobutyric acid

(GABA), although other targets such as the N-methyl D-aspartate (NMDA) receptor, μ

opioid receptor and CB1 cannabinoid receptor can also be important, depending on which

drug is involved. Skeletal muscle relaxants act peripherally at neuromuscular junction or in

the cerebrospinal axis to reduce muscle tone.

Acute toxicity studies showed mortality at 2500 mg/kg body weight, hence the

extracts were regarded safe to be administered at lower doses. Acute toxicity studies indicate

that petroleum ether extract of Amorphophallus paeoniifolius can be used safely up to doses

of 1500 mg/kg body weight. A spontaneous dose-dependent decrease in central nervous

system activity was found. The central nervous system depressants induce sedation and

reduce the locomotor activity of the experimenting animal. The mechanism of this depression

is not clearly understood at this point, but it can be assumed that the drug may exert central

nervous system depressant effect by interfering with the function of the cortex. Neither the

muscle relaxant activity has been understood.

Table 5. Data obtained for calculation of lethal dose (LD50)

Group Dose(mg/Kg) Log dose Dead/

Total

%Dead %Corrected Probit

1 1500 3.176 0/10 0 2.5 3.04

2 2000 3.301 2/10 20 20 4.16

3 2500 3.397 5/10 50 50 5.00

4 3000 3.477 8/10 80 80 5.84

5 3500 3.544 10/10 100 97.5 6.96

The peak central nervous system depression activity was observed at 60 min. The

recovery time of CNS depressant activity of petroleum ether extract (1000 mg/kg) and

diazepam (1.5 mg/kg) were found to be 24 hr and 20 hr, respectively14.



Table 6. Recovery of CNS depressant activity:

Time(h) Control group Group receiving

extract

Group receiving

Diazepam

0 177.5±24.85 122.5±29.17 119.5±22.39

1 176±23.35 25.25±3.77 22.75±3.30

4 - 32.25±10.07 30.5±4.86

8 - 52.5±14.52 46.25±9.81

12 - 65.25±10.62 66.25±10.01

16 - 79±5.35SS 90.75±7.76

18 - 91±12.67 105±17.33

20 - 108±8.75 120±18.67

22 - 120±7.76 -

24 - 122.75±23.94 -

CONCLUSION:

Herbal plants usually have great importance in the world of Pharmaceuticals. The

plant Amorphophallus paeoniifolius is found to be a potent herbal plant showing diverse

activities. The ethanolic extract shows antioxidant and antitumor activities. The leaves found

to have antidiarrhoeal activity. The effects of petroleum ether extract on the CNS activity of

the mice were observed.

Much money, time and energy have been spent in this field of research. So to get the

fruit of this labour, it is better to increase the re-evaluation and also further list of herbal

drugs have to be evaluated. So, further research has to be carried out to overcome the

shortcomings of the drugs and to relieve the suffering humanity and to obtain superior

therapeutically active drugs from this source.

REFERENCES:

1. http://wiki.answers.com/Q/What_is_the_importance_of_herbal_plants

2. S De, YN Dey, and AK Ghosh, Phytochemical investigation and chromatographic

evaluation of the different extracts of tuber of Amorphophallus paeoniifolius (Araceae),

International Journal on Pharmaceutical and Biomedical Research., Vol.1(5), pp.150-

157,2010.

3. KJ Bibu, AD Joy, and KA Mercey, Therapeutic effect of ethanolic extract of Hygrophila

spinosa T. Anders on gentamicin-induced nephrotoxicity in rats, Indian Journal of

Experimental Biology., Vol.48, pp.911-917, 2010.

4. Pratibha Singh, Manmohan Srivastava, and Lakhu Dev khemani, Renoprotective effects

of Andrographis paniculata (Burm f) Nees in rats, Upsala Journal of Medical Sciences.,

Vol.114, pp.136-139, 2009.

5. http://en.wikipedia.org/wiki/Amorphophallus

http://wiki.answers.com/Q/What_is_the_importance_of_herbal_plants

http://en.wikipedia.org/wiki/Amorphophallus



6. Shankhajit De, Yadu Nandan Dey, and Ajoy Kumar Ghosh, Anti-inflammatory activity of

methanolic extract of Amorphophallus paeoniifolius and its possible mechanism,

International Journal of Pharma and Biosciences., Vol.1(3), pp.1-8, 2010.

7. http://www.pfaf.org/user/Plant.aspx?LatinName=Amorphophallus+paeoniifolius

8. Yadu Nandan Dey, and Ajoy Kumar Ghosh, Pharmacognostic evaluation and

Phytochemical analysis of the tuber of Amorphophallus paeoniifolius, International

Journal of Pharma Research and Development., Vol.2(9), pp.44-49, 2010.

9. K Jagadheesh, V Arumugam, N Elangovan, and P Pavan Kumar, Evaluation of Anti-

tumor and Antioxidant Activity of Amorphophallus paeoniifolius on DMBA Induced

Mammary Carcinoma, International Journal of Chemical and Pharmaceutical Sciences.,

Vol.1(2), pp.40-50, 2010.

10. L Purwal, V Shrivastava, and UK Jain, Studies on Anti-diarrhoeal activity of leaves of

Amorphophallus paeoniifolius on experimental animals, International Journal of

Pharmaceutical Sciences and Research., Vol.2(2), pp.468-471, 2011.

11. Yadu Nanden Dey, Shankhajit De, Ajoy Kumar Ghosh, Sudhesh Gaidani, Suman

Kumari, and Mahvish Jamal, Synergistic depressant activity of Amorphophallus

paeoniifolius in Swiss albino mice, Journal of Pharmacology and Pharmacotherapeutics.,

Vol.2(2), pp.121-123, 2011.

12. Yadu Nandan Dey, Shankhajit De, and Ajoy Kumar Ghosh, Evaluation of analgesic

activity of methanolic extract of Amorphophallus paeoniifolius tuber by tail flick and

acetic acid induced writhing response method, International Journal of Pharma and

Bioscience., Vol.1(4), pp.662-667, 2010.

13. J Angayarkanni, KM Ramkumar, T Poornima, U Priyadarshini, Cytotoxic Activity of

Amorphophallus paeoniifolius Extracts Invitro, American-Eurasian Journal of

Agricultural and Environmental Sciences., Vol.2(4), pp.395-398, 2007.

14. SS Das, Malini Sen, YN Dey, S De, and AK Ghosh, Effects of Petroleum Ether Extract of

Amorphophallus paeoniifolius on Central Nervous System in Mice, Indian Journal of

Pharmaceutical Sciences., Vol.71(6), pp.651-655, 2009.

http://www.pfaf.org/user/Plant.aspx?LatinName=Amorphophallus+paeoniifolius

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