gjrmi - volume 3, issue 7, july 2014
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Global Journal of Research on Medicinal plants & Indigenous medicine - July 2014 issueTRANSCRIPT
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INDEX – GJRMI - Volume 3, Issue 7, July 2014
MEDICINAL PLANTS RESEARCH
Micro-biology & Bio-Chemistry
PHYTOCHEMICAL ANALYSIS AND ANTI-LIPID PEROXIDATION ACTIVITY OF TAMARIX
AFRICANA L. EXTRACTS
BENABDALLAH Hassiba, GHARZOULI Kamel, KHENNOUF Seddik, AMIRA Smain, SOUFANE Sihem
278–285
Bio-Technology
DIRECT SOMATIC EMBRYOGENESIS FROM MATURE LEAVES OF PIGEON PEA (CAJANUS
CAJAN L. MILL SP)
Pagadala Vijaya kumari 286–293
INDIGENOUS MEDICINE
Ayurveda - Kaumarabhritya
A COMPARATIVE STUDY OF BHASMAKNASHAK YOGA WITH EXERCISE AND DIET
RESTRICTIONS IN OVERWEIGHT CHILDREN
Renu B Rathi, Bharat Rathi
294–302
Ayurveda - Review Article
HEPATOPROTECTIVE HERBS USED IN AYURVEDA - A REVIEW
Giby Abraham
303–311
COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – INFLORESCENCE OF SAPTALA – ACACIA CONCINNA (WILLD.) DC.,
OF THE FAMILY MIMOSACEAE PLACE – KOPPA, CHIKKAMAGALUR DISTRICT,
KARNATAKA, INDIA
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 278–285
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
PHYTOCHEMICAL ANALYSIS AND ANTI-LIPID PEROXIDATION
ACTIVITY OF TAMARIX AFRICANA L. EXTRACTS
BENABDALLAH Hassiba1*, GHARZOULI Kamel
2, KHENNOUF Seddik
3,
AMIRA Smain4, SOUFANE Sihem
5
1Département de Microbiologie et Biochimie, Faculté des Sciences. Université de M’sila. Algérie.
2,3,4Département de Biologie, Faculté des Sciences de la Nature et de la Vie. Université de Sétif 1.
Algérie. 5Département de Biologie, Université de B.B. Arreridj. Algérie.
*Corresponding Author: E-mail: [email protected]
Received: 17/06/2014; Revised: 05/07/2014; Accepted: 06/07/2014
ABSTRACT
The homogenate from rabbit brain represents an important source of lipids used directly in the
study of peroxidation. The ratio of the peroxides is usually expressed as equivalent of
malondialdehyde and determined by using 1,1,3,3-tetramethoxypropane as standard. Tamarix
africana L. is widely used as a medicinal plant in Algeria. Polyphenols present in this plant are
considered active compounds. The extraction of the flavonoids of Tamarix africana L. allowed their
separation into two fractions (ethyl acetate extract and aqueous extract) containing flavonoids. The
effect of extracts of Tamarix africana L. was studied in vitro. Examination of the data showed a
significant inhibition of the relative rate of peroxidation by the ethyl acetate extract and the aqueous
extract in comparison with the control representing 100% of peroxidation. Since there was no
significant difference between these two extracts, the average rate is 53.8% inhibition. The ability of
extracts to reduce the rate of lipid peroxidation resulted mainly because of the presence of flavonoids
and phenolic acids.
KEYWORDS: flavonoids, lipid peroxidation, malondialdehyde, Tamarix africana L., thiobarbituric
acid.
Research Article
Cite this article:
BENABDALLAH Hassiba, GHARZOULI Kamel, KHENNOUF Seddik,
AMIRA Smain, SOUFANE Sihem (2014), PHYTOCHEMICAL ANALYSIS AND ANTI-LIPID
PEROXIDATION ACTIVITY OF TAMARIX AFRICANA L. EXTRACTS,
Global J Res. Med. Plants & Indigen. Med., Volume 3(7): 278–285
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 278–285
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
In addition to conventional drugs used in
the treatment of several diseases, traditional
medicine practiced in the world makes use of
plants to high levels of protective substances.
Among these plants, Artemisia herba alba L.,
Punica granatum L., Quercus ilex L. and
Tamarix africana L. which are widely used in
Algerian traditional medicine in the treatment
of gastroduodenal diseases. These plants are
rich in polyphenols (phenolic acids, tannins and
flavonoids) (Khennouf et al., 2003). Phenolic
compounds are secondary metabolites widely
distributed in the plant kingdom. Phenolic
acids, tannins and flavonoids are the major
classes of polyphenols. These compounds have
remarkable biochemical and pharmacological
activities such as antibacterial, antiviral and
anti-inflammatory activities (Kim et al., 1998;
Di Pietro et al., 2002). These activities are
mainly due to their antioxidant power (De
Whalley et al., 1990; Morton et al., 2000).
Lipid peroxidation leads to oxidative
degradation of unsaturated fatty acids. It
involves the reaction of the latter with
molecular oxygen to form a lipid radical and
semi-stable hydroperoxides (Tapel, 1973;
Barber and Bernheim, 1976). Many phenolic
compounds react with free radicals (Lonchampt
et al., 1989; Bagchi et al., 1998) to prevent the
degradation of membrane phospholipids which
is due to intense reactivity of free radicals
(Halliwell et al., 1992). Lipid peroxidation can
be enzymatic or non-enzymatic and occurs in
three stages: initiation, propagation and
termination (Halliwell and Gutteridge, 1984).
In Algeria, leaves of Tamarix africana L.
are traditionally used in decoction and infusion
in the treatment of disorders of the digestive
tract. In order to find principle compounds
responsible for this effect and to search the
mechanisms involved in this treatment, the
present study was conducted to extract the
flavonoids of this plant and to study their
effects on lipid peroxidation.
MATERIALS AND METHODS
The materials for the study were collected
during June in the region of Bordj Bou
Arreridj, Algeria during the fruiting period. The
plant was taxonomically identified using flora
of Quézel and Santa (1962–1963), Ozenda
(1983) and Maire (1952–1987); verified,
characterized and confirmed by professional
botanists of the department. Voucher
specimens were deposited in the Herbarium.
The samples of plant (leaves) were cleaned and
allowed to dry at room temperature. The dried
material is ground for use in the extraction of
flavonoids.
Extraction of flavonoids
The extraction of flavonoids was performed
according to the method recommended by
Markham (1982). It was based on the degree of
solubility of these compounds in organic
solvents. This method has two major steps: the
first is with methanol to dissolve the flavonoids
and the second is with chloroform and ethyl
acetate to separate aglycones and glycosylated
fractions of flavonoids. The extraction of
flavonoids of Tamarix africana L. is made
from the finely ground dry matter. After two
successive extractions with 85% and 50%
methanol, the filtrates were subjected to
evaporation at low pressure (35°C Vapor Rota,
Büchi 461, Germany). The filtrate was freed of
waxes, fats and chlorophyll by successive
washings with n-hexane to give an aqueous
phase. To separate aglycones flavonoids and
glycosylated flavonoids, the aqueous phase was
mixed with chloroform to obtain an organic
phase containing the flavonoid aglycones and
aglycones methoxylated. The remaining
aqueous phase underwent a series of
extractions with ethyl acetate to recover the
organic phase which contained some flavonoid
aglycones, but especially mono- and
diglycosides flavonoids. The remaining
aqueous phase contained more polar
glycosylated flavonoids such as di-, tri- and
tetraglycosides flavonoids. In this study, two
extracts were used: ethyl acetate extract and
aqueous extract. The collected fractions were
submitted to a concentration at low pressure at
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 278–285
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
35°C and then lyophilized for 24 hours. Each
lyophilisate was weighed to calculate the yield
of the extraction, expressed in grams per 100
grams of freeze-dried matter.
Determination of total polyphenols
The determination of total polyphenols was
performed according to the method of Prussian
blue and Price Butler (1977), modified by
Graham (1992). The concentration of total
polyphenols was derived from a standard curve
prepared with gallic acid as the standard. The
assay results were expressed as milligrams
equivalent of gallic acid per gram of freeze-
dried matter.
Determination of total flavonoids
The quantitative determination of
flavonoids was performed according to the
method of aluminium trichloride (Bahorun et
al., 1996). A standard curve was set separately
with quercetin to calculate the concentration of
flavonoids in each extract. Assay results were
expressed in milligrams equivalent of quercetin
per gram of freeze-dried matter.
Lipid peroxidation
Both extracts of Tamarix africana L. (ethyl
acetate extract and aqueous extract) were
suspended in the 0.5% carboxymethylcellulose
(CMC) at final concentrations 10, 25 and 50
μg/ml. The CMC solution alone was used as a
control solution.
The measure of the level of lipid
peroxidation was carried out on a rabbit brain
homogenate. Animals were anesthetized by
intraperitoneal injection of urethane (25%).
Cold saline (0.9% NaCl) was infused through
the jugular vein to the brain to rid the blood of
the tissue. The brain was removed and
homogenized in 1.15% KCl.
The rate of peroxide was measured by the
method described by Ohkawa et al., (1979).
This method was based on the reaction between
peroxides and thiobarbituric acid (TBA) which
lead to the formation of a pink complex
indicator of lipid peroxidation. The brain
homogenate were incubated at 37°C in a water
bath for one hour in the presence or absence of
the test solutions. At the end of the incubation
period, the mixture was centrifuged at 2700 g
for 10 minutes (Rotina 35R, Hettich, Germany)
and the supernatant were added to 8.1% sodium
dodecyl sulfate (SDS) and 0.8% TBA prepared
in acetic acid. The mixture was heated in a
water bath at 100°C for one hour. After
cooling, the samples were subjected to a second
centrifugation to eliminate proteins and the
optical density of the supernatant was read at
532 nm. The results are expressed as relative
rate of peroxidation in relation to peroxidation
of the homogenate untreated (control).
Relative rate of peroxidation (%) =
(absorbance of sample/absorbance of control) ×
100.
Chemicals
Aluminium trichloride, acetic acid, gallic
acid, CMC, quercetin, TBA, MDA, SDS were
of analytical grade (Fluka, Merck, Prolabo,
Sigma).
Statistical Analysis
The results of different experiments are
expressed as mean ± SEM. The calibration
curves were calculated by the method of linear
regression. The significant difference between
control and treated groups was determined by
analysis of variance on ranks followed by
Dunn's test for multiple comparisons with
α=5%.
RESULTS
Extraction and determination of total
phenolic compounds
Extraction of flavonoids by organic
solvents from dry weight of Tamarix africana
L. showed that the aqueous extract is the
highest (19–22%) compared with ethyl acetate
extract (3–4.5%) (Table 1).
The determination of total polyphenols by
the modified Prussian blue method showed the
sensitivity and the reproducibility of this
method. The amounts of total phenolic
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 278–285
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
compounds in the two extracts of Tamarix
Africana L. are shown in Table 1. A high
content was observed in ethyl acetate extract
(500–546 mg/g) in comparison with the
aqueous extract (140–170 mg/g).
Aluminium trichloride is a method used for
the determination of total flavonoids. Content
of flavonoids is expressed as mg quercetin per
g of freeze-dried matter. The amount of
flavonoids is higher in ethyl acetate extract
(332.2 mg/g) in comparison with the aqueous
extract (14.2 mg/g).
Effect of extracts on lipid peroxidation
Incubation of the homogenate of rabbit
brain for an hour in the presence of extracts of
Tamarix africana L. (ethyl acetate extract and
aqueous extract) showed that these extracts (10,
25 and 50 µg/ml) inhibited lipid peroxidation.
The presence of one of the two extracts in the
incubation medium induced a significant
decrease in the relative rate of peroxidation.
Since there is no significant difference between
these two extracts, which give 53.8% of
inhibition of lipid peroxidation (Fig. 1).
Table 1. Determination of the rate of different classes of phenolic compounds of Tamarix
africana L. extracts.
Extracts Yield (%) Total polyphenols*
(mg/g)
Total flavonoids**
(mg/g)
Ethyl acetate
3.0–4.5
500–546
332
Aqueous 19.0–22.0 140–170 14
* Modified Prussian blue method.
** Aluminium trichloride method.
Figure 1. Effect of ethyl acetate and aqueous extracts of Tamarix africana L. on the rate of lipid
peroxidation.
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 278–285
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
DISCUSSION
Folk medicine practiced throughout the
world relies heavily on the use of plants as a
source of natural active substances. Among
these substances, plants containing phenolic
compounds and flavonoids in particular are
highly beneficial in therapeutics. Starting from
the observation that Tamarix africana L. is
widely used in Algerian traditional medicine
and to obtain a good separation of their active
principles responsible for its gastroprotective
activity, the mining method adopted is based on
differences in the degree of solubility of
flavonoids in organic solvents. A relatively
high yield (19–22%) was obtained with the
aqueous extract of Tamarix africana L.
containing the most polar flavonoids (di-, tri-
and tetraglycosides). In contrast, the
performance of the ethyl acetate extract
containing some flavonoid aglycones including
mono- and diglycosides is five times lower than
that of the aqueous extract.
Several methods are applied to extracts for
the quantitative determination of different
classes of phenolic compounds of Tamarix
africana L. (total polyphenols and flavonoids).
The modified Prussian blue method was
effective for the determination of total
polyphenols in the extracts. The quantitative
determination of phenolic compounds showed
that the amount of polyphenols in the ethyl
acetate extract is relatively high in comparison
with the aqueous extract. Quercetin is widely
used as a standard for determining the content
of flavonoids in a sample. In comparison with
the ethyl acetate extract, the aqueous extract
appears poor in flavonoids.
Analysis of extracts of Tamarix africana L.
with HPLC (unpublished results) showed the
presence of quercetin, kaempferol, luteolin and
isorhamnetin in the ethyl acetate extract. In
addition, the aqueous extract contains mainly
flavonoid glycosides or rutinosides and some
flavonoid aglycones which isoquercetin and
luteolin are identified by this method. In
addition to flavonoids, phenolic compounds
identification by HPLC showed that the
extracts contain procyanidins and some
phenolic acids such as ellagic acid, gallic acid
and vanillic acid. These data confirm that the
extraction process adopted is acceptable to
some extent to separate flavonoid glycoside
and aglycone. The richness of Tamarix
africana L. on active compounds such as
flavonoids, phenolic acids and tannins can be
one of the principles of its use in traditional
medicine in the treatment of diseases of the
digestive tract.
Lipid peroxidation leads to oxidative
degradation of unsaturated fatty acids and leads
to the alteration of the structural integrity of
membranes and their permeability. However,
the conversion of Thiobarbituric Acid Reactive
Substances (TBARS) equivalent of MDA is
widely used to assess the importance of lipid
peroxidation (Wills, 1987; Minamiyama et al.,
1994). In the present study based on the
extinction coefficient of MDA, the incubation
of tissue for an hour results in the formation of
11.6 ± 1.5 nmol MDA equivalents per gram of
fresh tissue. This rate is not far from that found
in the liver homogenate (11.9 ± 1.1 nmol/g)
and rat stomach (20.0 ± 2.5 nmol/g) (Yegen et
al., 1990). Flavonoids, phenolic acids and
tannins inhibit mechanisms of enzymatic and
non-enzymatic initiation of lipid peroxidation
(Nakayama et al., 1992; Galvez et al., 1995;
Morton et al., 2000). Incubation of rabbit brain
homogenate in the presence of caffeic acid,
gallic acid and ellagic acid showed inhibition of
lipid peroxidation. This result is consistent with
that of Okuda et al., (1992) and Nardini et al.,
(1998) who found that phenolic acids are
powerful antioxidants, including those with a
catechol -type structure such as caffeic acid.
The gallic acid and caffeic acid have the same
antiradical efficiency (Sanchez –Moreno et al.,
1998). The esterification of caffeic acid and
gallic acid increases their antiperoxidation
activity (Nakayama et al., 1992).
The application of two extracts of Tamarix
africana L. in peroxidation test revealed their
antioxidant potential. The ethyl acetate extract
and the aqueous extract have a similar effect.
As mentioned before, the two extracts contain
some aglycones but mostly glycosylated
flavonoids. The presence of flavonoids and
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 278–285
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
phenolic acids in Tamarix africana L. extracts
suggests that the mechanisms involved in the
antioxidant activity of these extracts are the
same as those of the pure phenolic compounds
(scavengers of free radicals, chelating metals).
CONCLUSION
Extraction protocol applied for the
separation of flavonoids of Tamarix africana L.
into two fractions is acceptable and it has
achieved in all quite acceptable extraction
yields. Moreover, the quantitative
determination of different classes of phenolic
compounds showed that the ethyl acetate
extract is rich in flavonoids. Ethyl acetate
extract and aqueous extract inhibited lipid
peroxidation in vitro. Although the two
fractions are quite complex in their
composition, they exert a similar effect. As the
phenolic compounds have other activities in
addition to their antioxidant effect, they can
replace the classic antioxidant, ascorbic acid.
The fact that the composition of each extract is
complex, it is necessary to isolate and assess
the major active principles of this plant to test
their effect against several diseases.
ACKNOWLEDGEMENT(S)
The National Agency for the Development
of Health Research and the Ministry of Higher
Education and Research Scientist of Algeria are
thanked for financial support for research
projects (07/01/01/03/06/97, F/1901-04-95).
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Source of Support: National Agency for the
Development of Health Research and the
Ministry of Higher Education and Research
Scientist of Algeria
Conflict of Interest: None Declared
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 286–293
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
DIRECT SOMATIC EMBRYOGENESIS FROM MATURE LEAVES OF
PIGEON PEA (CAJANUS CAJAN L. MILL SP)
Pagadala Vijaya Kumari1*
!Department of Botany, Cytogenetics and Plant Biotechnology Laboratory, Osmania University,
HYDERABAD - 500 007, India.
Present Address: Biotechnology, Department of Biology, Ambo University , AMBO – Ethiopia.
*Corresponding Author: Email: [email protected]
Received: 16/05/2014; Revised: 20/06/2014; Accepted: 30/06/2014
ABSTRACT
Protocols were standardized for plant regeneration via direct somatic embryogenesis from 35-
day-old leaf explants of three cultivars of pigeonpea [Cajanus cajan] (ICPH-8, ICPL-87 and ICPL-
7295). Frequency of Somatic embryo induction was dependent on the age of the leaves. Leaves
isolated from in vitro and glass house grown plants responded well when compared to field grown
plants. Leaves produced embryogenic calli from cut ends and somatic embryos appeared directly
from leaf margins when cultured on MS medium supplemented with 5 mg/l naphthalene acetic acid
(NAA), 1 mg/l, 6-benzylaminopurine (BAP) and 6% sucrose. Dark incubation of cultures for 30
days showed a remarkable increase in frequency (50) of somatic embryogenesis. Somatic embryos at
various developmental stages matured upon transfer to 0.2 mg/l BAP and 0.1 mg/l NAA with 4%
sucrose in half-strength MS medium. Plantlets obtained from somatic embryos were transferred to
pots for acclimatization and grown to maturity with 70–80% frequency.
KEY WORDS: Somatic embryogenesis; Mature leaf; Pigeon pea; Cajanus cajan; grain legume.
Research Article
Cite this article:
Pagadala Vijaya kumari (2014), DIRECT SOMATIC EMBRYOGENESIS FROM MATURE
LEAVES OF PIGEON PEA (CAJANUS CAJAN L. MILL SP), Global J Res. Med. Plants &
Indigen. Med., Volume 3(7): 286–293
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 286–293
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
Pigeon pea (Cajanus cajan (L Mill sp) is
one of the major grain legume (pulse) crops of
the tropics and subtropics. Direct somatic
embryogenesis is the formation of somatic
embryos or embryo-genic tissue directly from
the explant without the formation of an
intermediate callus phase (Raghavan 1986). In
embryogenesis systems, this is almost always
what happens (Merkle et al., 1990; Finer
1994). Unfortunately, in most other plants
somatic embryogenesis is more difficult to
obtain, but progress has been made in some
grain legumes. Grain Direct embryogenesis
occurs when embryos are started directly from
explant tissue creating an identical clone while
indirectly occurs from unorganized tissue
(callus). Plant transformations and
Mass propagation
are the two important
methods which can make use of these Somatic
Embryogenesis. Agrobacterium mediated
transformation is the easiest and most simple
plant transformation can be done by using
these somatic embryogenesis. legumes are
difficult to regenerate, however different few
reports are available from different explants
where the concept of regeneration was
observed. Anbazhagan and Ganapathi (1999)
described a protocol for initiation of cell
suspension cultures from leaflet explants and
subsequent plant regeneration via somatic
embryogenesis in Cajanus cajan. haploid plant
production by anther culture (Bajaj et
al.,1980). An efficient and reproducible
protocol for regeneration is an urgent need to
develop transgenics and genetic variations in
Pigeon pea. Few reports on direct somatic
embryogenesis in Pigeon pea from different
explants and genotypes are available.
Regeneration of shoot buds from excised
cotyledons of Pigeon pea with BAP was
reported earlier (Kumar et al., 1983; Mehta U
& Mohan Ram., 1980). Plant regeneration was
obtained on MS medium with IAA, Kinetin
and coconut mild in Pigeon pea (Patel et al.,
1982). The frequency of success on in vitro
plant regeneration via somatic embryogenesis
is very low (Seenivasu et al.,1998) All the
protocols reported till date were callus
mediated, and the present report mainly deals
with direct somatic embryogenesis from
leaves. For the first time, high frequency plant
regeneration via direct somatic embryogenesis
in Pigeon pea from mature leaf margins was
accomplished. This finding is of great
significance in developing the transformation
protocols, which may be exploited in Pigeon
pea crop improvement.
MATERIALS AND METHODS
Plant material - Seeds of three pigeonpea
genotypes ICPH-8, ICPL-87 and ICPL-7295
were procured from Genetic Resources Unit,
International Crops Research Institute for Semi
- arid Tropics, Patancheru, and Hyderabad,
India. Seeds were surface sterilized with 70%
ethanol for 5 minutes followed by 0.1%
aqueous mercuric chloride for 10 min. They
were washed thoroughly with sterile water and
germinated aseptically on hormone free
Murashige and Skoog’s (1962) (MS) medium
containing 3% sucrose and 0.8% difco bacto
agar. MS medium containing 4–6% sucrose,
BAP, NAA, indole-3-acetic acid (IAA), 2, 4-
dichlorophenoxyacetic acid (2, 4-D), zeatin
(ZEA) and kinetin (KN) either alone or in
combination was used for induction of somatic
embryos and maturation. The pH of all the
media was adjusted to 5.8 before autoclaving.
The media were dispensed into culture tubes of
25×150 mm and sterilized at 1.4 Kg cm2 for 20
minutes.
Experimental design / Methodology - The
effect of age of the leaf on somatic embryo
induction was investigated by collecting them
from 5 to 40 day-old seedlings grown in field
(29–35°C), in vitro (25°C) and glass house
(30°C). Leaf number was counted from the top
after the initiation of first two pair of leaves
from the seedlings and the age from the date of
emergence was noted. In all the cases, 25
explants from each set were inoculated. The
effect of growth regulators such as BAP, NAA,
2,4-D, ZEA, IAA and KN (0.5–10.0 mg/l)
either alone or in combinations was studied on
the induction of somatic embryos. All the
experiments were repeated twice with ten
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 286–293
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replicates, frequency of somatic embryogenesis
and standard deviation were calculated.
Induction of somatic embryos - Mature
leaves aged 35-days were cut vertically into
two pieces and inoculated onto MS medium
supplemented with 6% sucrose, 5 mg/l NAA
and 1.0 mg/l BAP. The cultures were incubated
for three weeks at 25 2 C under light (60
Em-2
s-1
) with 16 / 8 light/dark photoperiods.
The calli along with proembryos on the leaf
margins were subcultured into the same media
and incubated for 30 days in dark. Globular
and heart shaped embryos were separated and
transferred onto half-strength MS medium
supplemented with 4% sucrose, 0.2 mg/l BAP
and 0.1 mg/l NAA. Plantlets with well-
developed roots were transferred to
experimental pots with 75% humidity for
hardening and then to greenhouse for
acclimatization.
Histological studies - For histological
observations, the leaf along with callus and
somatic embryos was fixed in acetic acid:
ethanol (1:3) for 72 hours, dehydrated in an
ethanol and butanol series followed by paraffin
embedding as described by Sharma & Sharma
(1980). The embedded tissue was cut with
microtome (Reichert-Jung, No 2030 Supercut,
Germany) into 10 m thick sections, stained
with hematoxylin eosin, mounted with DPX
and observed microscopically.
RESULTS AND DISCUSSION
The frequency of somatic embryogenesis
was highest (70%) in 35-day-old leaf explants
with an average of 52 somatic embryos per
explant in presence of 5 mg/l NAA and 1 mg/l
BAP only when incubated in dark for 30 days
(Fig. 1A). Prolonged incubation in dark for 35
and 42 days decreased the frequency of
embryogenesis to 20% and 10% respectively.
Thus, it appeared that light may not be
necessary for the induction of somatic
embryogenesis. Similarly, in Triticale, (1)
reported 2.25-fold increase in embryoid
induction in dark as compared to light
incubation at (3000 lux). At the end of third
week, the leaf margins showed tiny, globular,
smooth, greenish as well as golden yellow
coloured proembryos in bunches. Embryogenic
callus was also induced from the cut ends of
the leaf. Culture of 40 day-old leaf resulted
only in callus initiation without somatic
embryogenesis. This implies that age of the
leaf is critical for embryo induction. Explants
collected from in vitro grown seedlings
responded well compared to leaf of other
seedlings (Table 1).
In the presence of different concentrations
of auxins and cytokinins, callus was induced
without somatic embryogenesis when cultures
were incubated in dark for 30-days. At the
concentrations tested, 2, 4-D yielded golden
yellow, KN brownish yellow and ZEA green
calli (Table 2). The effect of auxin along with
cytokinin at various concentrations was tested
for the induction of somatic embryos from the
cultivar ICPH-8 (Table 3). With a NAA (5
mg/l), BAP (1 mg/l) and 6% sucrose the
frequency of somatic embryogenesis was 70%
only when incubated in dark for 30-days.
Increasing concentrations of sucrose increased
the frequency of response until 6% and
declined thereafter (Fig. 1B). After three – four
weeks, calli along with proembryos on the leaf
margins were subcultured into the same media
and incubated for 28–30 days in dark. Different
stages of somatic embryos appeared only
during this second subculture (Fig. 2A).
Histological sections of these cultures revealed
the presence of embryos at different stages (all
the pictures of the embryos not shown)(Fig.
2B&C). They were isolated and transferred to
half-strength MS medium fortified with 0.1
mg/l NAA, 0.2 mg/l BAP and 4% sucrose for
maturation. Embryos developed into plants
between 3–4 weeks on this medium (Fig. 2 D
& E). Plantlets with well-developed roots were
transferred into experimental small pots for 10
days for hardening at 75% humidity and
26 1 C and subsequently to green house for
acclimatization (Fig F). Induction of somatic
embryos in pigeonpea via callus from different
explants with NAA and BAP, failed to develop
further into complete plants (Nalini et
al.,1996).
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Table 1. Effect of source on leaf Somatic embryogenesis in Pigeonpea Cv.ICPH-8.
Age of
leaf(D)
Field grown
plants
Glass house
grown plants
In vitro grown
plants
10 0 0 0
25 0 C C+S.E
35 0 C+S.E C+S.E
40 0 C C+S.E D-Days, C-Callus, S.E-Somatic Embryogenesis
Table 2. Effect of Auxins and Cytokinins on the induction of callus in Pigeonpea Cv. ICPH-8.
Auxins Cytokinins
2,4-D IAA NAA BAP KN ZEA
(mg/l) % (mg/l) % (mg/l) % (mg/l) % (mg/l) % mg/l) %
0.5 50 0.5 60 1.0 20 0.5 30 1.0 20 0.5 20
1.0 10 1.0 0 2.0 0 1.0 0 2.0 0 1.0 0
2.0 15 2.5 20 2.5 0 2.0 0 2.5 0 1.5 0
2.5 17 2.5 0 5.0 0 2.5 0 5.0 0 2.0 0
3.0 20 3.0 0 6.0 0 3.0 10 6.0 0 2.5 0
4.0 25 4.0 0 7.5 0 3.5 15 7.5 0 3.0 20
5.0 30 5.0 0 10.0 0 5.0 0 10.0 20 3.5 0
6.0 29 7.5 0 0 0 6.0 0 0 0 5.0 25
7.5 40 0 0 0 0 7.5 0 0 0 6.0 40
10.0 20 0 10 0 0 10.0 5 0 0 10.0 0 % = % Frequency of callus induction
Table 3. Effect of auxin and cytokinin on the induction of somatic embryogenesis from leaf
margins of Pigeonpea Cv.ICPH-8
Conc.growth
Regulators (mg/I)
Type of Response % of Somatic
embryogenesis
Average No. of
somatic embryos
2,4-D 2.0 + KN 0.5 C 0 0
2,4-D 5.0 + KN 0.5 C 0 0
2,4-D 7.5 + KN 1.0 C 0 0
2,4-D 2.0 +BAP 1.0 C 0 0
2,4-D 5.0 + BAP0.5 C 0 0
2,4-D 7.5 +BAP 1.0 C 0 0
NAA 1.0 + BAP 1.0 C 0 0
NAA 2.0 + BAP 1.0 C 0 0
NAA 3.0 + BAP 1.0 C 0 0
NAA 4.0 + BAP 1.0 C + S.E 25 ± 1.5 15 ± 0.7
NAA 5.0 + BAP 1.0 C + S.E 70 ± 2.1 52 ± 1.5
NAA 6.0 + BAP 1.0 C + S.E 30 ± 0.9 20 ± 1.2
Values represent mean SD ( C-Callus, S.E - Somatic embryos )sucrose 4%
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Table 4. Effect of BAP and NAA on maturation of Somatic embryos of Pigeonpea CV. ICPH 8
Hormones (mg/l) % of S.E. matured No.of S.E/explants
BAP 0.5 + 0.1 NAA 0 0
BAP 1.0 + 0.5 NAA 0 0
BAP 0.5 + 0.5 NAA 10 ± 0.5 5 ± 0.6
BAP 1.0 +1.0 NAA 0 0
BAP 0.2 + 0.1 NAA 65 ± 1.2 39 ±1.3
BAP 0.2 + 0.5 NAA 15 ± 0.9 10 ±0.2
Values represent mean SD, S.E Somatic embryos
1A - Age of the leaf corossponding with frequency of somatic embryos.
1B – Sucrose Concentration showing the percentage of Somatic Embrygenesis.
1C - Genotype Variation with percentage of Somatic Embryos.
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 286–293
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In the present study, some of the embryos
germinated on the leaf explant giving rise to
shoots with two small leaflets with out roots.
Anatomical studies in pigeonpea, made from
different explants (Prakash et al., 1994)
showed the initiation of shoot buds from sub-
epidermal tissues. In the present study also,
epidermal tissue of the leaf margins might have
become competent for embryo induction.
Different concentrations of BAP and NAA
were tried for maturation of embryos and their
subsequent germination into plantlets. Mature
embryos were separated and transferred onto
half-strength MS with 0.2 mg/l BAP and
0.1 mg/l NAA (Table 4). The frequency of
maturation of embryos and subsequent
conversion into plantlets was 65% with shoot
and root average of (39) somatic embryos per
explant. The response of other two cultivars
ICPL-87 and ICPL-7295 were also evaluated
for the induction of somatic embryos (Fig. 1C)
and the frequency if somatic embryogenesis
was similar. These observations suggest that
induction of embryogenesis from leaf explants
may be genotype independent (Hazra et al.,
1989). Induction of somatic embryogenesis in
groundnut was reported by (Bernard., 1980)
with 2,4-D alone or 2,4-D plus NAA. Somatic
embryogenesis in pigeonpea was observed
earlier by (George & Eapen., 1994), while they
used 2,4-D, NAA or picloram. On the other
hand (Seenivasu et al., 1998) used thidiazuran,
but in all the above reports somatic
embryogensis were observed in callus cultures.
2A 2B 2C
2D 2E 2F 2 A. Mature leaf margins showing bunch of tiny globular stage somatic embryos.
2 B. Histological section of a globular embryo.
2 C. Histological section of globular embryo with suspensor from the margins of mature leaf.
2 D. Young plantlet germinated from bipolar embryo with shoot and root systems
2 E. Plants with well-established shoot and roof system derived from somatic embryos.
2 F. Regenerated Plant in the Experimental Pot.
Figure 2:
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 286–293
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
CONCLUSION
The present study clearly suggests that
direct somatic embryogenesis from matured
leaf in all the three cultivars of Pigeonpea were
induced with high frequency (60–70%) for the
first time. The Plantlets were regenerated and
transferred to pots and were grown to maturity
with high frequency 70–80%. The cultivar
ICPH–8 with MS medium, 6% sucrose gave
the maximum frequency of embryogenesis and
also regeneration. Further standardization for
the plantlet survival to soil is under
experimentation. This finding has great
significance and can be successfully exploited
in developing genetic variations including
transgenics of this economically important
crop.
ACKNOWLEDGEMENTS
The author is grateful to Dr. Ramanandam,
ICRISAT, Patanchuru for providing the seeds
and to Dr. Shashikaran, National Institute of
Nutrition (NIN), Hyderabad for carrying out
histology (microtomy) work. Senior Research
Fellowship awarded by the Council of Scientific
and Industrial Research - CSIR, New Delhi, to
Ms Pagadala Vijaya Kumari is gratefully
acknowledged. Heartful thanks to Prof
J.K.Bhalla (Ex- Head Dept of Botany) for
constant encouragement and Supervision in
Manuscript preparation. Together with
Department of Biology - AMBO University
Ethiopia for their constant Co operation.
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Bajaj, Y.P.S., Singh, H., Gosal, S.S. (1980).
Haploid embryogenesis in cultures of
Pigeon pea ( Cajanus cajan) Theor.Appl.
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hexaploid triticale: determination of
physical consitions increasing embryoid
and green plant production.
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Dixon RA, Gonzales RA (eds) Plant
cell culture: a practical approach.
Oxford University Press, Oxford, pp
67–102
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Kumar AS, Reddy TP & Reddy GM, Plant
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cultures of pigeonpea (Cajanus cajan
L).Plant Sci. Lett , 32 (1983) 271.
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plants from cotyledons of cajanus cajan.
Indian J. Exp. Biol , 8 (1980) 800.
Merkle SA, Parrott WA, Williams EG (1990)
Applications of somatic embryogenesis
and embryo cloning. In: Bhojwani SS
(ed) Plant tissue culture: applications
and limitations. Elsevier, Amsterdam,
pp 67–102
Murashige T & Skoog F, A revised mediun for
rapid growth and bioassay with tobacco
tissue culture Physiol. Plant, 15 (1962)
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Nalini Mllikarjuna, Reena MJT, Sastri DC &
Moss JP, Somatic embryogenesis in
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Exp. Biol,4 (1996) 282.
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Patel, D.B., Barve, D.M., Nagar, N., Mehta,
A.R. In vitro development of immature
and hybrid embryos of cajanus cajan L)
(1994). Indian J. Exp. Biol. 32: 740–
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Prakash NS, Pental D & SarinNB, Regeneration
of pigeon peas ( Cajajus cajan) from
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formation. Plant Cell Rep, 13 (1994)
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Raghavan V (1986) Embryogenesis in
angiosperms: a developmental and
experimental study. Cambridge
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Seenivasu K.,Malik SK, Ananda Kumar P&
Sharma RP, Plant regenratin via
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Source of Support: NIL Conflict of Interest: None Declared
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 294–302
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
A COMPARATIVE STUDY OF BHASMAKNASHAK YOGA WITH EXERCISE
AND DIET RESTRICTIONS IN OVERWEIGHT CHILDREN
Renu B Rathi1*
, Bharat Rathi2
1Professor & Head, Kaumarbhritya Department, MGACHRC, Salod, Wardha, Maharashtra, India
2Dr. Bharat Rathi, M.D. Professor & Head, Rasshastra Bhaishajya Kalpana Department, MGACHRC, Salod,
Wardha, Maharashtra, India
* Corresponding Author: E-Mail: [email protected]; 919011058302
Received: 29/05/2014; Revised: 25/06/2014; Accepted: 01/07/2014
ABSTRACT
Due to modern lifestyle, the rate of developing overweight in children is also increasing
tremendously. It is a proved fact that exercise and diet restriction of specially sweet and fatty
foodstuffs help to reduce weight, maintain fitness and can have the direct effect to prevent systemic
illness. In overweight individuals, there will be increased hunger and thirst which again facilitates
weight gain. In Rasatantrasaara and Siddhaprayoga Sangraha, it has been mentioned that
Apamarga seeds (Achyranthus aspera Linn.) are useful in Bhasmaka Vyadhi (Voracious appetite).
The action of Apamarga seeds was considered to be hard to digest creates sense of fullness and
enables to cope with overweight. The study aims at evaluating the efficacy of Bhasmaknashak yoga
in overweight children as compare to exercise and diet restrictions. Total 30 patients of age 8–15
years were divided into 3 groups of each 10. Subjects of Group A were put on exercise and diet
restriction, Group B was administered with trial drug and Group C received both. It shows that the
statistical effect on clinical features in group C was highly significant than group A and B (P values-
BMI <0.0005, Abdominal circumference <0.0005, climbing time <0.0001). The study revealed that
awareness of overweight problems and adaptation of change in lifestyle along with remedies are
much important in treating overweight as compare to remedy or regimen alone.
KEY WORDS: Bhasmaknashak yoga, overweight, exercise and diet restrictions.
Research Article
Cite this article:
Renu B Rathi, Bharat Rathi (2014), A COMPARATIVE STUDY OF
BHASMAKNASHAK YOGA WITH EXERCISE AND DIET RESTRICTIONS
IN OVERWEIGHT CHILDREN, Global J Res. Med. Plants & Indigen. Med.,
Volume 3(7): 294–302
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 294–302
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
In recent years, there is an increasing trend
of developing overweight in children. There are
lots of reasons behind it like more consumption
of fast food, bakery items, sedentary life style
and lack of outdoor playing. There is 16.9%
prevalence of obesity in children and
adolescents aged between 2–15 years (Ogden et
al., 2012). Obesity is associated with a
multitude of adverse health effects. Central or
visceral fat in obesity pours out free fatty acids
and increases insulin resistance. The adipose
cells secrete multiple hormones, known as
‘adipokines,’ and markers of inflammation (H.
M. Chandola, H. Sharma, 2013). According to
WHO, a BMI greater than or equal to 25 is
overweight and a BMI equal or more than 30 is
obese (Centers for Disease control &
prevention, 2014). The term most commonly
used to quantify overweight is Body Mass
Index or BMI calculated as Wt in kg / Ht in m2.
It is a proved fact that exercise and diet
restrictions play a key role in reducing
overweight (Thomas A. et al., 2005), as
‘exercise’ means = designed, repetitive for the
rationale of training any part of the body, hence
taken for the study in group A. Drug
intervention used in present study
‘Bhasmaknashak yoga’ (Krishnanand &
Badrinarayan Shastri, 1991) is widely used in
clinics to treat voracious appetite. Seeds of
Achyranthus aspera are proved to have
properties like anti-obesity potential (Neerja
Rani et al., 2012) hepatoprotective
(Manjunatha BK, et al., 2012; Kokila Parmar,
et al., 2013) etc. Till date there is no
documented clinical study on trial drug, as per
author’s knowledge, hence the study was taken
up with an aim of evaluating the therapeutic
efficacy of ‘Bhasmakanashka Yoga’ in
overweight children. The study was also
planned in order to evaluate the role of diet
restrictions specially fried, sweet food and
adoption of regular exercise or outdoor playing
in reducing the overweight in children and to
see the combined effect of remedy and regime.
MATERIALS AND METHODS:
Study design and duration
It is an open ended randomized
comparative clinical pilot study.
Total 30 patients were divided into 3 groups
randomly as per liking and consent. 30 patients
of Sthaulya (overweight) attending the OPD of
Balrog, Mahatma Gandhi Ayurveda College,
Hospital and Research Centre, Wardha,
Maharashtra, India were registered for this
study. Present study was a pilot study with
small sample size, further in continuation, a
project with large sample size has been
submitted for ethical approval.
Group A – All 10 overweight children
receiving diet instructions (Diet chart not to eat
sweets, fried food stuffs was given to them) and
at least half an hour daily exercise or outdoor
play.
Group B - All 10 overweight children
receiving trial drug Bhasmaknashak yoga in a
dose of 3gm with hot water twice a day post
meal up to 1 month.
Group C - All 10 overweight children were
instructed for taking orally Bhasmaknashak
yoga with diet restrictions and exercise.
Drug Review: The prime ingredients of this
yoga have been enumerated in Table I. All
drugs were taken in equal proportion, powdered
and mixed together to prepare the trial
formulation.
Dose and Duration: 3 gm twice a day, with
hot water, post meal for a period of one month
Follow-up has been taken up to 1 month and
found that drug effect existed in.
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 294–302
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Table I showing ingredients of the Trial drug Bhasmaknashak yoga with Sanskrit and
Botanical names with citation.
Inclusion Criteria:
All overweight children of 8 to 15 years age
group having > 25 BMI
Child who was not taking weight-affecting
medications directly or indirectly.
Child not having a medical condition for
which a weight loss program would be
contraindicated.
Exclusion Criteria:
Overweight children having hormonal
disorders like hypothyroidism and Diabetes
mellitus.
Genetic disorders like Down’s syndrome,
Turner syndrome.
Kwashiorkar, Nephrotic syndrome,
Congestive Cardiac Failure in which there
is anasarca, overviewed as overweight (Lily
John, D.B. Tripathi, 1994).
Withdrawal Criteria: The children who had
left the treatment modality during the course or
being irregular.
Assessment Criteria:
1. Weight, height, BMI were recorded before
starting the treatment and later on every week
of the study. Weight was also recorded for all
the patients who have come for the follow-up
study.
2. Abdominal circumferences at umbilicus
level were recorded before and thereafter every
week, till the completion of the course of
treatment, to assess the effect of therapy.
3. Climbing time: The time taken to climb fixed
number of stairs ten continuously without
taking rest in seconds was taken as climbing
time to assess the clinical features like Guruta,
Ayasen Shwas, Daurbalya.
No investigations had been studied like Lipid
profile, etc. due to fund constraint as it was a
pilot study.
Statistical Analysis-was done by Sigma state
software, paired t test
RESULTS :
It was observed that maximum patients
were belonging from the age group of 10 to 15
years. Male patients were dominant total 24
boys (80%) and 6 girls in this study in all the 3
groups. The main cause of overweight was over
eating in 73.33%, very less physical exertion in
60% and heredity cause was observed in 90%
(At least 1 parent was overweight) in all the 3
groups. The common clinical features were
similar to medoroga like desire for more food,
Sr. No. Sanskrit Name Botanical Name Useful part
1 Amalki Phyllanthus emblica Linn. Fruit
2 Bibhitaki Terminalia belerica Roxb. Fruit
3 Haritaki Terminalia chebula Retz. Fruit
4 Musta Cyperus rotundus Linn. Rhizome
5 Apamarga Achyranthus aspera Linn Seeds
6 Pippali Piper longum linn Fruit
7 Vidanga Embelia ribes Burm Fruit
8 Sita Sugar -
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water, cool air, sleep etc. (KR Srikantha
Murthy, 2002).
The symptoms found in all the children
enrolled for the study had Guruta
(90%), Nidraadhikya-oversleep (86.34%),
Atipipasa – over thirst (93.97%), AtiKshudha-
voracious appetite (100%). Other signs and
symptoms observed were
Daurbalya (92.77%), Ayasena shvasa
(91.6%), Utsahahani (92.77%), Atisvedapravr
uti (82.15%). It provided significant relief
in Utsahahani (30.58%), Daurbalyata (28.41),
Anga Guruta (25.67%), Nidradhikya
(23.88%) and Aayasena Shvasa (19.4%),
Atiswedpravruti with statistically highly
significant (p<0.0001 in all features) in group C
children. Along with clinical features the
results were drawn by using paired t test to
judge the effect by different parameters like
BMI, Abdominal circumference and climbing
time which has been depicted in the following
tables (Tables II to XIII).
Table-II, showing statistical efficacy on clinical features in Group A
Clinical Feature* Mean D Std. Error t value p value
BT AT BT AT
Atikshudha 3.26 2.57 0.69 0.070 0.068 10.24 <0.0001
Atitrushna 3.090 2.500 0.59 0.057 0.073 7.42 <0.0001
Atinidra 3.378 2.689 0.69 0.040 0.061 14.2 <0.0001
Guruta 3.220 2.350 0.87 0.119 0.096 5.54 <0.0005
Utsahhani 3.470 2.320 1.15 0.076 0.116 7.01 =0.0001
Aayasen Shwas 3.350 2.590 0.76 0.054 0.087 8.86 <0.0001
Daurbalya 3.370 2.610 0.76 0.068 0.092 9.28 <0.0001
Atisweda 3.390 2.840 0.55 0.071 0.044 5.83 =0.0003
* Atikshudha-Voracious appetite, Atitrushna-over thirst, Atinidra-Over sleep, Guruta-
heaviness, Utsah hani-lethargic, Aayasen shwas- dyspnea/palpitation, Daurbalya- Weakness,
Atisweda- over sweating
Table-III, showing statistical efficacy on clinical features in Group B
Clinical Feature Mean D Std. Error t value p value
BT AT BT AT
Atikshudha 3.05 2.52 0.53 0.060 0.065 12.2 <0.0001
Atitrushna 3.040 2.520 0.52 0.068 0.093 9.390 <0.0001
Atinidra 3.311 2.900 0.41 0.067 0.041 10.65 <0.0001
Guruta 3.280 2.410 0.87 0.120 0.056 7.292 <0.0001
Utsahhani 2.850 2.250 0.60 0.0764 0.099 9.000 <0.0001
AayasenShwas 3.350 2.8300 0.52 0.054 0.059 6.868 =0.0001
Daurbalya 3.230 2.760 0.47 0.055 0.045 10.48 <0.0001
Atisweda 3.180 2.821 0.36 0.059 0.062 7.962 <0.0001
P value<0.0001-highly significant
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Table -IVshowing statistical efficacy on clinical features in Group C
Clinical Feature Mean D Std. Error t value p value
BT AT BT AT
Atikshudha 3.54 1.620 1.92 0.091 0.093 18.63 <0.0001
Atitrushna 3.760 2.180 1.58 0.077 0.063 15.66 <0.0001
Atinidra 3.34 2.47 0.87 0.05 0.047 21.34 <0.0001
Guruta 3.400 1.400 2.00 0.047 0.058 24.91 <0.0001
Utsahhani 3.170 1.530 1.64 0.059 0.083 15.68 <0.0001
AayasenShwas 3.71 2.35 1.36 0.038 0.070 26.12 <0.0001
Daurbalya 3.760 1.690 2.07 0.035 0.127 15.62 <0.0001
Atisweda 3.530 1.432 0.21 0.049 0.029 19.23 <0.0001
P value<0.0001-highly significant
Table V, showing effect on BMI in all groups-
Group Mean D Std. Error t
value
p
value BT AT BT AT
A 20.55 19.39 1.16 0.0957 0.216 4.824 <0.001
B 21.39 19.40 1.950 0.192 0.339 5.802 <0.001
C 24.04 20.39 3.65 0.297 0.434 12.632 <0.001
P value<0.0001-highly significant
Table VI, showing Effect on Abdominal circumference in all groups-
Group Mean D Std. Error t value p value
BT AT BT AT
A 84.530 81.870 2.260 0.409 0.265 5.854 <0.001
B 85.27 82.36 2.910 0.146 0.475 7.472 <0.001
C 90.09 84.80 5.29 0.736 0.342 7.952 <0.001
P value<0.001-highly significant
Table VII, showing Effect on climbing time in all groups-
Group Mean D Std. Error t value p value
BT AT BT AT
A 17.8 16.4 1.32 0.218 0.243 5.381 <0.0001
B 18.88 17.86 1.00 0.322 0.395 3.348 <0.0001
C 19.38 14.87 4.51 0.228 0.241 21.31 <0.0001
P value<0.001-highly significant
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Table-VIII, showing comparison of means in Clinical features of group A Versus B
Clinical Feature D DF Std. Error t value p value
Atikshudha −0.05 18 0.09402 −0.532 =0.6014
Atitrushna −0.25 18 0.11812 −2.117 <0.0005
Atinidra −0.48 18 0.8898 −2.117 <0.0005
Guruta 0.06 18 0.11127 0.539 =0.5963
Utsahhani −0.07 18 0.1528 −0.458 =0.6523
AayasenShwas 0.24 18 0.10274 2.336 =0.0313
Daurbalya −0.48 18 0.08898 −5.394 <0.0001
Atisweda −0.48 18 0.08898 −5.394 <0.0001
Table-IX, showing comparison of means in objective parameters in group A versus B
Objective Criteria D DF Std. Error t value p value
BMI 0.1 18 0.4015 0.249 0.8061
Abd. Circumference 0.49 18 0.5443 0.900 0.3799
Climbing Time 0.30 18 0.305 0.767 0.453
Table-X, showing comparison of means in clinical features of group B Vs C
Clinical Feature D DF Std. Error t value p value
Atikshudha 0.9 18 0.11306 7.961 <0.0001
Atitrushna 0.34 18 0.11214 3.032 =0.0072
Atinidra 0.17 18 0.0611 2.783 =0.0123
Guruta 1.01 18 0.0809 12.484 <0.0001
Utsahhani 0.72 18 0.12937 5.566 <0.0001
AayasenShwas 0.48 18 0.08898 5.394 <0.0001
Daurbalya 0.48 18 0.08898 5.394 <0.0001
Atisweda 0.48 18 0.08898 5.394 <0.0001
P value<0.0001-highly significant
Table XI, showing comparison of means in objective parameters in group B versus C
Objective Criteria D DF Std. Error t value p value
BMI 0.99 18 0.5502 1.799 <0.0005
Abd. Circumference 2.44 18 0.5853 4.169 <0.0005
Climbing Time 3.49 18 0.093 9.408 <0.0001
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Table -XII, showing comparison of means in group C versus A
Clinical Feature D DF Std. Error
t value p value
Atikshudha 0.95 18 0.11532 8.238 <0.0001
Atitrushna 0.32 18 0.09637 3.320 <0.0005
Atinidra 0.219 18 0.07478 2.929 <0.0005
Guruta 1.01 18 0.0809 8.496 <0.0001
Utsahhani 0.95 18 0.11182 5.529 <0.0001
AayasenShwas 0.48 18 0.08898 5.394 <0.0001
Daurabalya 0.48 18 0.05838 5.134 <0.0001
Atisweda 0.48 18 0.08898 5.394 <0.0001
P value<0.0001-highly significant
Table-XIII, showing comparison of means in objective parameters in group C versus A
Objective Criteria D DF Std. Error t value p value
BMI 1.09 18 0.4842 2.251 <0.0005
Abd. Circumference 2.93 18 0.4325 6.775 <0.0001
Climbing Time 3.19 18 0.033 9.847 <0.001
P value<0.0001-highly significant
DISCUSSION:
Individually all the three modalities of
overweight treatment were significant in
lowering chief complaints as well as objective
parameters but integrative approach has more
quick and long lasting efficacy over separate
regimen or remedy as seen in follow-up period.
It was found that individually every group has
statistically significant in clinical features but
while comparing the effect of exercise and diet
restrictions versus trial drug between Group A
and B, it was observed that Group B was
effective than group A in decreasing some chief
complaints like Daurbalya and Atisweda which
proving that Group B had good impact over
Group A as a treatment modality of overweight
children.
While comparing the effect of trial drug
versus integrative approach between Group B
and C, it was observed that Group C was highly
effective than group B in decreasing almost all
chief complaints except Atitrushna and
Atinidra. Also C group has statistical
significance on objective criteria on climbing
time which proving that Group C has good
impact over Group B as a treatment modality of
overweight children. While comparing the
effect of therapy between Group C and A, it
was observed that Group C was highly
effective than group A in decreasing almost all
chief complaints like Guruta, Utsah hani,
Aayasen Shwas, Daurbalya and Atisweda. Also
C group had statistical significance on
objective criteria like abdominal circumference
and climbing time which proves that Group C
had an edge over Group A as a treatment
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modality of overweight children. The mean
BMI, Abdominal circumference and climbing
time of group C children were less as compare
to group A and B. All children felt easiness in
routine activities with increased strength,
decrease in dyspnea post treatment
intervention.
Ingredients of Bhasmaknashakyoga have
effect on overweight due to their Kapha-
vathara (normalizing body humour) lekhana,
(scrapping of body fat) deepana, pachana
(normalizing metabolism) properties (Priyavrat
Sharma, 2013; G. Pande, 2013).
Mishri/sita provides palatability to the
formulation. It was observed that the
formulation was safe and had no any untoward
effect in group B and C children. Group B and
C children had their meal as routine. After
study, children were advised for Psychologist’s
counseling to decrease the affinity for diet.
In this study, 4 patients were withdrawn
due to irregularity in treatment modalities and
replaced with other 4 overweight children.
Follow-up study- After completion of due
course of treatment, all the children were asked
to report for follow-up study for a period of 1
month. In follow-up study significant changes
in body weight, abdomen circumference were
observed.
CONCLUSION:
Overweight is becoming a burning problem
in society. Above study reveals that to
overcome it, multi-dimensional approach is
essential. It’s not so easy and instant to change
ones personality that too in children. The study
has shown that awareness of overweight and
adaptation of change in lifestyle along with
remedies are much important in its treatment.
Here as compare to remedy or regimen alone,
the combination of both as in group C children
shown more benefits when compared to group
A and B.
From the study, it can be concluded that
trial medicine having encouraging weight
reducing effect but it can be enhanced by
adding diet restrictions and exercises. This was
a pilot study, but it’s encouraging results
indicate the need of further extensive research
with large sample and long term follow-up.
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Source of Support: NIL Conflict of Interest: None Declared
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 303–311
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
A REVIEW ON HEPATO-PROTECTIVE HERBS USED IN AYURVEDA
Giby Abraham1*
1Research and Development, Confederation for Ayurvedic Renaissance – Keralam Limited (CARe Keralam
Ltd.), KINFRA Small Industries Park, Nalukettu Road, Koratty, Thrissur – 680 309, Kerala, India
*Corresponding Author: E-mail:[email protected]; Mob: 09995215790
Received: 10/06/2014; Revised: 20/06/2014; Accepted: 25/06/2014
ABSTRACT
Liver is considered to be one of the vital organs which helps in maintaining the health of body.
Yakrit (liver) is being described right from the vedic period. Modern lifestyles can overstress the
liver and make it malfunctioning. No significant and safe hepato-protective drugs are available in
modern therapeutics. The nature has bestowed some plants with the property to prevent, treat and
cure hepatic disturbances with interception of fewer side effects. The focus of this review is to
elucidate the importance of liver and aimed at compiling data based on reported works on medicinal
plants that have been tested in hepato-toxicity models and proved as hepato-protective. Also the
probable mode of action of a few herbs has been discussed in Ayurvedic and modern aspect.
KEY WORDS: Liver, Yakrit, Hepato-protective, Medicinal plants.
Review Article
Cite this article:
Giby Abraham (2014), A REVIEW ON HEPATO-PROTECTIVE HERBS USED IN
AYURVEDA, Global J Res. Med. Plants & Indigen. Med., Volume 3(7): 303–311
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 303–311
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
"Is your life worth living”? It depends on
the liver as it is the largest glandular organ in
the body which works all the time to keep the
body healthy. The liver is important because a
person’s nutritional level is not only
determined by what he or she eats, but by what
the liver processes. The incredible complexity
of liver chemistry and its fundamental role in
human physiology is so daunting to researchers
that the thought that simple plant remedies
might have something to offer is astonishing
and incredible!
Liver is considered to be one of the most
vital organs that functions as a centre of
metabolism of nutrients such as carbohydrates,
proteins and lipids and excretion of waste
metabolites. Additionally, it is also handling
the metabolism and excretion of drugs and
other xenobiotics from the body thereby
providing protection against foreign substances
by detoxifying and eliminating them.
In Ayurvedic literature, yakrit (liver) is
considered as an important anga (organ) of the
human body right from the vedic period.
Bhavamisra (16th
Century) has described that it
is situated right and below to the hridaya
(heart) and is the sthana (seat) of pitta and
sonitha (blood) (Srikantha Murthy, 2002).
Susrutha (500 BC) mentioned yakrit (liver) as
the abode of ranjaka pitta (Srikantha Murthy,
2004).
Susrutha (500 BC) describes yakrit (liver)
as the sthana (seat) of rakta (blood) (Srikantha
Murthy, 2005). Charaka (1000 BC) while
describing the srotas (body channels),
mentioned yakrit (liver) and pleeha (spleen) as
the moola (root) of raktavaha Srotas (blood
carrying channels) (Sharma and Dash, 2007).
But it is Bhavamisra who for the first time
introduced the term ‘yakrit vikara’ (liver
disorders). Madhavanidana, in parishista
prakarana, explains yakrit roga (liver disease)
as a separate entity (Yadunandan Upadhyaya,
2000). The etio-pathogenesis of Yakrit roga has
been described in Figure 1.
In dealing with problems of the liver, the
primary goal is to enhance liver detoxification
processes and to help protect against further
liver damage. Significant and safe hepato-
protective agents are unavailable in modern
therapeutics. Therefore, due importance has
been given globally to develop plant-based
hepato-protective drugs effective against a
variety of liver disorders.
The present review is aimed at compiling
data based on reported works on promising
phytochemicals from medicinal plants that have
been tested on hepato-toxicity models.
Figure 1: Etio-pathogenesis of yakrit roga (liver disease)
Vidahi annapanam (food and
drinking that cause burning
sensation, Madya sevana
(alcohol intake),
Teekshna padartha
(strong/ sharp substance)
Pitta prakopa
(aggrevation of pitta)
Dushita rakta dhatu
(vitiated blood tissue)
Rakta pradoshaja
roga (disease
caused by vitiated blood tissue)
yakrit-pleeha-
kamala roga (diseases of
liver, spleen,
jaundice etc.)
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Table 1. showing some hepato-protective herbs with their pharmacological properties
(Sharma P.V, 2009)
S.N Plant
Rasa
Guna Virya Vipaka Dosakarma
1. Guduchi
(Tinospora cordifolia
(Willd.) Miers.)
Tikta,
Kashaya
Guru,
Snigdha
Ushna Madhura Tridoshahara
2. Pippali
(Piper longum Linn.)
Katu Laghu,
Snigdha,
Teekshna
Ushna Madhura Tridoshahara
3. Punarnava
(Boerhavia diffusa Linn.)
Madhura,
Tikta,
Kashaya
Laghu,
Ruksha
Ushna Madhura Tridoshahara
4. Kalamegha
(Andrographis paniculata
Nees.)
Tikta Laghu,
Ruksha
Ushna Katu Kaphapittahara
5. Bhumyamalaki
(Phyllanthus niruri Linn.)
Tikta,
Kashaya,
Madhura
Laghu,
Ruksha
Seeta Madhura Kaphapittahara
6. Daruharidra
(Berberis aristata DC.)
Tikta,
Kashaya
Laghu,
Ruksha
Ushna Katu Kaphapittahara
7. Katuki
(Picrorhiza kurroa Royle
ex Benth.)
Tikta Laghu,
Ruksha
Seeta Katu Kaphapittahara
8. Rohitaka
(Techoma undulata G.
Don.)
Katu,
Tikta,
Kashaya
Laghu,
Ruksha
Seeta Katu Kaphapittahara
9. Bhringaraja
(Eclipta alba Hassk.)
Tikta,
Kashaya
Laghu,
Ruksha
Ushna Katu Kaphavatahara
10. Sharapunkha
(Tephrosa purpurea Pers.)
Tikta,
Kashaya
Laghu,
Ruksha,
Teekshna
Ushna Katu Kaphavatahara
[Rasa (taste) – Katu (pungent), Tikta (bitter), Kashaya (astringent)
Guna (quality) – Guru (difficult to digest), Snigdha (unctous) Laghu (easily digestible), Ruksha (dry), Teeksha (sharp)
Virya (potency) – Seeta (cold), Ushna (hot)
Vipaka (post metabolic effect)
Dosa karma (action on functional entites of the body), hara (pacifies)]
Hepatoprotective Drugs
The important herbs used in Ayurveda to treat
Liver diseases have been described in Table 1.
Tinospora cordifolia (Willd.) Miers.
(Guduchi)
Tinospora cordifolia (Willd.) Miers.,
known as Guduchi, Amrita is one of the most
valuable medicinal herbs of Ayurveda. The
term 'Amrita' is attributed to this herb in
recognition of its ability to impart youthfulness,
vitality and longevity to its patron. In modern
medicine, it is well known for its
hepatoprotective, adaptogenic, immuno-
modulatory activities and anti-fibrolytic
activity. The active principle Tinosporin
corrects immunosuppression associated with
deranged hepatic function (Varsha et al., 2011).
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Kupffer cells are major determinants of
outcome of liver injury. The effect of
Tinospora cordifolia (Willd.) Miers. was
evaluated on Kupffer cell function, using
carbon clearance test as a parameter.
Antihepatotoxic activity of Tinospora
cordifolia (Willd.) Miers. was studied in albino
rats intoxicated with Carbon tetrachloride
(CCl4). Liver function was assessed based on
morphological, biochemical (SGPT, SGOT,
Serum alkaline phosphatase, Serum bilirubin)
and functional (Pentobarbitone sleep time)
tests. A study conducted by Nagarkatti et al.,
(1994) on Tinospora cordifolia (Willd.) Miers.
indicates that it had decreased fibrosis in rats,
induced by CCl4 and significantly improved the
suppressed Kupffer cell function in another rat
model of chronic liver damage induced by
heterologous serum. This raises the possibility
that anti-fibrotic effect of Tinospora cordifolia
is mediated through activation of kupffer cells.
Piper longum Linn. (Pippali)
Piper longum Linn. belongs to the family
Piperaceae, is a common Indian dietary spice
which has been shown to possess a wide range
of therapeutic utilities. It has been reported to
possess antiasthmatic, antiinflammatory,
hepatoprotective, hypocholestremic and
immunomodulatory activities. It contains
various alkaloids like piperine, piperlongumine,
piperlonguminine, etc. which helps in the
regeneration of hepatocytes (Gupta AK, 2003).
A study conducted by Jagruti and Urvi
(2009) showed a significant hepatoprotective
effect on Piper longum Linn. milk extract
treatment in CCl4 induced hepatic damage. An
evident decrease in level of serum enzymes,
total bilirubin and direct bilirubin was
observed. Histo-pathological findings indicated
that administration of Piper longum Linn. milk
extract offered protection to the hepatocytes
from damage induced by CCl4, with mild fatty
changes in the hepatic parenchymal cells,
which corroborated the changes observed in the
hepatic enzymes. It also showed regenerating
liver cells around the necrotic area.
Boerhavia diffusa Linn. (Punarnava)
The roots of Boerhavia diffusa Linn.,
commonly known as 'Punarnava', are used by a
large number of tribes in India for the treatment
of various hepatic disorders and for internal
inflammation. Clinical data has also reported
effectiveness of Boerhavia diffusa Linn. in
cases of oedema and ascites resulting from
early cirrhosis of the liver and chronic
peritonitis (Varsha et al., 2011). The effect of
ethanolic extract of roots of Boerhavia diffusa
Linn. on country made liquor induced
hepatotoxicity was studied in albino rats by
Agarwal et al.(1991). Histo-pathological
studies showed marked reduction in fat
deposits in animals receiving Boehavia diffusa
Linn. along with country made liquor. The
plant protected the rats from hepatotoxic action
by decreasing the serum alanine amino
transferase (ALT), triglycerides, cholesterol
and total lipid levels in both serum and tissues.
Punarnava contains alkaloids named as
punarnavine and punarnavoside which shows
anti-fibrinolytic activity but the
hepatoprotective activity has been attributed to
ursolic acid. Keppler and co-workers
demonstrated that ursolic acid isolated from the
leaves showed a dose dependent (5–20 mg/kg)
hepatoprotective activity (21–l00%) in rats
against thioacetamide, galactosamine and
carbon tetrachloride induced hepatotoxicity in
rats. These hepatotoxins decreased the viability
of hepatocytes as assessed by trypan blue
exclusion and rate of oxygen uptake tests and
decreased the volume of bile as well as the
level of its contents. Pretreatment with ursolic
acid increased the viability of rat hepatocytes
significantly (Keppler et al., 1968).
Andrographis paniculata Nees. (Kalamegha)
Andrographis paniculata Nees. is an
ancient Indian medicinal herb, which has been
used for centuries in Asia for its effects on
various bodily functions and ailments, ranging
from degenerative diseases to the common
cold. The plant is known as King of Bitters.
Andrographolide is an active constituent
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 303–311
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extracted and isolated from Andrographis
paniculata Nees which is very bitter in taste
(Anil Kumar et al., 2012).
A study conducted by Visen et al. (1993)
on andrographolide showed a significant dose
dependent protective activity against
paracetamol-induced toxicity on ex vivo
preparation of isolated rat hepatocytes. It
significantly increased the percent viability of
the hepatocytes as tested by trypan blue
exclusion and oxygen uptake tests. It blocked
the toxic effects of paracetamol on certain
enzymes (GOT, GPT and alkaline phosphatase)
in serum as well as in isolated hepatic cells. The bioactive constituent also antagonizes toxic
effects of CCl4 and acetaminophen on certain
enzymes (GOT, GPT and alkaline phosphates)
in serum as well as in isolated hepatic cells.
The results clearly depicted the plant extract to
exert a choleretic effect that reduces the
cholestasis and diminishes retention as well as
increase the excretion of toxic xenobiotics from
liver. Further, it also stimulated immune system
to fight against inflammation, is mediated from
the release of cytokinin from
immunomodulators (Varsha et al., 2011).
Phyllanthus niruri Linn. (Bhumyamalaki)
Phyllanthus niruri Linn. is a medicinal herb
used in connection with secondary hepatitis and
other ailments, in ayurvedic medicine for over
2000 years.
It is a proved antiviral drug in Hepatitis-B
in human subjects. In the preliminary study,
carriers of Hepatitis-B virus were treated with a
preparation of the plant 200 mg for 30 days. 22
of the 37(59%) treated patients had lost
Hepatitis-B surface antigen, when tested 15–
20days after the end of the treatment, compared
with only 1 out of 23 (4%) placebo treated
controls. It has exhibited an inhibition of DNA
polymerase on Hepatitis–B virus which is
responsible for the replication of virus
(Mehrotra et al., 1991).
In a study, phyllanthin, hypophyllanthin
and tricotanol were isolated from petroleum
ether extract of Phyllanthus niruri Linn. shows
significant results on rat hepatocytes.
Preclinical studies demonstrate that an extract
of the Phyllanthus niruri Linn. plant inhibits
endogenous DNA polymerase of hepatitis B
virus and binds to the surface antigen of
Hepatitis B virus. Extracts of Phyllanthus
niruri Linn. have been shown to exert
hepatoprotective effect against CCl4 induced
HepG2 cell damage in rabbits. Pre-treatment
with extract of Phyllanthus niruri Linn.,
reduced paracetamol-induced acute liver
damage in rats as monitored by estimating the
SGOT. In the in vitro-study, it decreased the
release of AST and ALT in rat primary cultured
hepatocytes being treated with ethanol
(Tabassum et al., 2005).
Berberis aristata DC. (Daruharidra)
Berberis asiatica DC. being an important
medicinal plant is used extensively for
treating variety of ailments like infection of
eyes, skin diseases, jaundice and rheumatism
(Kirtikar and Basu, 1933). The major
alkaloid of this plant is reported to be berberin
which possess anti-oxidant property (Brijesh
and Khosa, 2010).
The roots of Berberis aristata DC. possess
more effective hepatoprotective activity against
CCl4 intoxication in rats because of its
antioxidant bearing capacity. Acute CCl4
administration increased serum and liver lipid
peroxides significantly. Berberine treatment
could reduce these elevated levels. Pathological
analysis showed degeneration and necrosis
after CCl4 administration. Berberine treatment
could minimize these effects to a certain extent.
(Brijesh and Khosa, 2010)
Picrorhiza kurroa Royle ex. Benth (Katuki)
Picrorhiza kurroa Royle ex Benth. is a
renowned herb in the Ayurvedic system of
medicine and has traditionally been used to
treat disorders of the liver, upper respiratory
tract, reduce fevers, treat dyspepsia, chronic
diarrhoea, and scorpion sting. Kutkin, the
active principal of Picrorhiza kurroa Royle ex.
Benth is comprised of kutkoside and iridoid
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glycosides like picrosides I, II, and III
(Chaturvedi and Singh, 1996).
The hepato-protective action of Picrorhiza
kurroa Royle ex Benth. may be attributed to its
ability to inhibit the generation of oxygen
anions and to scavenge free radicals.
Picrorhiza’s antioxidant effect has been shown
to be similar to that of superoxide dismutase,
metal-ion chelators and xanthine oxidase
inhibitors. Animal studies indicate that
Picrorhiza’s constituents exhibit a strong anti-
cholestatic activity against a variety of liver-
toxic substances, appearing to be even more
potent than silymarin (Chander et al., 1992).
Picrorhiza also exhibits a dose-dependent
choleretic activity, evidenced by an increase in
bile salts and acids, and bile flow (Shukla et al.,
1991).
Techoma undulata G. Don (Rohitaka)
Techoma undulata G. Don is a tropical
coastal shrub that grows up to 1 m in height. It
occurs throughout the Indian subcontinent.
Techoma undulata G. Don leaves were tested
against liver damage of albino rats. Loss of
membrane structure and integrity because of
lipid peroxidation was accompanied with the
elevated levels of marker enzymes like SGOT,
SGPT and total bilirubin. This shows that the
plant has got membrane stabilizing function.
Techoma undulata G. Don was potentially
effective in blunting lipid peroxidation,
suggesting that the extract possibly has
antioxidant property to reduce ethanol-induced
membrane lipid peroxidation and thereby to
preserve membrane structure and might be due
to the presence of glycosides, flavonoids,
proteins, amino acids, tannins, saponins and
triterpenoids (Singh D. et al., 2011).
Eclipta alba Hassk. (Bhrngaraja)
Eclipta alba Hassk. known as Bhringraja,
is a plant belonging to the family Asteraceae. In
ayurvedic medicine, the leaf extract is
considered a powerful liver tonic. It possesses a
wide range of biological activities and is used
for the treatment of hepatitis and cirrhosis
(Wagner, H. 1986). A study conducted by
Murugaian P et al., 2008 on the whole plant
extract of Eclipta alba Hassk. exhibited the
protective activity against CCl4 induced liver
injury. The plant contains an alkaloid Ecliptine
which has got choleretic action. The extract
augmented the bile flow in rats suggesting a
stimulation of liver secretory capacity.
Tephrosia purpurea Pers. (Sharapunkha)
Tephrosia purpurea Pers. known as
Sharapunkha, forms one of the most effective
ingredients of formulations available in Indian
market for liver ailments. In the traditional
Indian medicine it is famous for its
effectiveness in bilious febrile attacks,
obstruction of liver and spleen apart.
Especially, it has shown good results in
cirrhosis and viral hepatitis in clinical trials
(human studies). Dried ethanolic extract of
Tephrosia purpurea Pers. was studied for its
efficacy using both acute and chronic models
CCl4 of experimentally induced hepatotoxicity.
In vitro studies exploiting trypan blue
exclusion assay revealed that the alcoholic
extract exerted a significant hydroxyl radical
scavenging activity (Sree Rama Murthy and
Srinivasan, 1993).
Hepato-protective effect of aerial parts was
evaluated against CCl4 induced hepatotoxicity
in rats. An oral dose of powdered aerial parts to
rats prevented the elevation of SGOT, SGPT,
Bilirubin levels caused by CCl4. The
mechanism of hepato-protection by Tephrosia
purpurea Pers. mainly involves membrane
stabilization of liver cells as indicated by
decrease in levels of SGOT, SGPT and
bilirubin levels, wherein it prevents cellular
leakage and loss of functional integrity of the
liver cell membranes caused by various
hepatotoxic agents. Tephrosia purpurea Pers.
also leads to increase in hepatic regeneration,
which again contributes to its hepatoprotective
efficacy (Jain, A. et al., 2006).
DISCUSSION
Yakrit (liver) is the sthana (seat) of pitta
dosha (functional entity of the body), rakta
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 303–311
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dhatu (blood tissue) and agni (power of
digestion). Treatment of all liver diseases in
Ayurveda concentrates mainly on pitta dosha
rather than the organ itself. Most of the hepato-
protective drugs are kapha pitta samaka
(pacifies pitta & kapha entities). The medicines
and diets that normalise pitta are commonly
used for all types of liver diseases.
Most of the hepato-protective herbs are
having predominantly Tikta- Katu rasa (bitter
and pungent taste) and deepana- pachana
karma (digestive stimulant and carminative).
These herbs are mainly agni vardhaka
(increases fire entity in the body) and act on
jatharagni (digestive fire) as well as dhatwagni
(fire residing in tissues). These rasas (taste)
have the property of increasing metabolism
(mainly enhancing catabolism), thereby these
herbs help in digestion of nitrogenous waste
products collected in body, due to disturbed
metabolism. Most of the hepato-protective
herbs possess laghu (easy to digest) and ruksha
(dry) gunas (quality). Laghu guna (easy to
digest quality) helps in increasing jatharagni
(digestive fire) as they are easily digestible and
they form less nitrogenous waste products.
Ushna virya (hot potency) help in enhancing
the Jatharagni (digestive fire) as well
dhatwagni because ushna virya (hot potency)
increases metabolism (catabolism).
According to modern pharmacology, the
main mechanism involved in the protection of
liver could be associated with the strong
capability of hepato-protective drugs to reduce
the intracellular levels of reactive oxygen
species by enhancing the level of both
enzymatic and non-enzymatic antioxidants.
These drugs protect liver tissues against
oxidative damage and somehow help in
stimulating the repair mechanism of liver.
The mode of action of hepato-protective
herbs varies from herb to herb. Hepatocyte
membrane stabilizing capacity is shown by
Techoma undulata G. Don., thereby preventing
toxins from entering the cell through entero-
hepatic recirculation. Berberis aristata DC.,
Tephrosa purpurea Pers. and Piper longum
Linn. help in regeneration of liver cells by
stimulating nuclear polymerase A and
increasing ribosomal protein synthesis.
Tinospora cordifolia (Willd.) Miers. enhance
the activity of Kuffer cells which is involved in
the production of substances like interleukins
and tumour necrosis factors which activate the
immune system of the body and act as
immuno-modulatory. Phyllanthus niruri Linn.
possess antiviral property and help in
microsomal induction or inhibition. Boerhavia
diffusa Linn possess antifibrinolytic activity.
Eclipta alba Hassk., Andrographis paniculata
Nees. and Picrorhiza kurroa Royle ex Benth.
increase the choleretic activity.
Different single herbs are very much useful
in liver disorders as shown by research studies.
A few Ayurvedic compound formulations such
as Phalatrikadi kwatha, Vasa guduchyadi
kashaya, Patola katurohinyadi kashaya, Guda
pippali, Arogyavardhini vati, Rohitakarista
mentioned in Sharangadhara Samhitha (13th
Century) are also found to be promising in
hepatopathy.
CONCLUSION
The challenge that modern medical system
face with liver disorders is that such drugs
would have to be metabolized in the liver.
Since the liver itself is in disorder, the problem
is how to ensure effective metabolism of the
drugs that have been prescribed. In this context,
Ayurveda sages have used their genius, to
formulate such herbal formulations that can be
metabolized even by a sluggish liver. The logic
on which such formulations work is that they
first heal and reinvigorate the liver and thus
contribute to the restoration of its normal
functions. Preserving health of the liver means
adding healthier years to one’s life. Be polite to
your liver & Keep it Living and Lively!!
ACKNOWLEDGEMENT
Author is thankful to Dr. Rajasekhara N,
Head & Professor and Dr. Vijayalakshmi P.B,
Lecturer, Dept. of Dravyaguna, KVG Ayurveda
College, Sullia for all the help and guidance in
writing this article.
Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 7 | July 2014 | 303–311
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