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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

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Renu B Rathi, Bharat Rathi

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Giby Abraham

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OF THE FAMILY MIMOSACEAE PLACE – KOPPA, CHIKKAMAGALUR DISTRICT,

KARNATAKA, INDIA

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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: benabdallahhassiba@yahoo.com

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: vkpagadala@rocketmail.com

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.

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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.

REFERENCES

Anbazhagan, V.R., Ganapati, A. (1999).

Somatic embryogenesis in cell

suspension cultures of pigeon pea

(Cajanus cajan). Plant Cell Tiss. Org.

Cult.56:179–184.

Bajaj, Y.P.S., Singh, H., Gosal, S.S. (1980).

Haploid embryogenesis in cultures of

Pigeon pea ( Cajanus cajan) Theor.Appl.

Genet. 58: 157–159.

Bernard S, (1980). Invitro androgenesis in

hexaploid triticale: determination of

physical consitions increasing embryoid

and green plant production.

Z.Pflanzenzuchtung, 85 : 308−321.

Finer JJ (1994) Plant regeneration via

embryogenic suspension cultures. In:

Dixon RA, Gonzales RA (eds) Plant

cell culture: a practical approach.

Oxford University Press, Oxford, pp

67–102

George L. & Eapen S, Organogenesis and

embryogenesis from diverse explants in

pigeon pea. Plant Cell Rep, 13 (1994)

417.

Hazra S, Sathaye SS & Mascarenhas, BioTech,

7(1989) 949.

Kumar AS, Reddy TP & Reddy GM, Plant

regeneration from different callus

cultures of pigeonpea (Cajanus cajan

L).Plant Sci. Lett , 32 (1983) 271.

Mehta U & Mohan Ram HY, Regeneration of

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)

473.

Nalini Mllikarjuna, Reena MJT, Sastri DC &

Moss JP, Somatic embryogenesis in

pigeon pea (cajanus cajan L ) Indian J.

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–

744.

Prakash NS, Pental D & SarinNB, Regeneration

of pigeon peas ( Cajajus cajan) from

cotyledonary node via multiple shoot

formation. Plant Cell Rep, 13 (1994)

623–627.

Raghavan V (1986) Embryogenesis in

angiosperms: a developmental and

experimental study. Cambridge

University Press, New York

Seenivasu K.,Malik SK, Ananda Kumar P&

Sharma RP, Plant regenratin via

somatic embryogenesis in pigeon pea (

Cajanus cajan ( L.) Mill sp). Plant Cell

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Source of Support: NIL Conflict of Interest: None Declared

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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: rbr.226@gmail.com; 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.

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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.

REFERENCES

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(2014),

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Bhavprakash Nighantu, Chaukhambha

Bharati Academy, Reprint edition, pg

no 400

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Ayurveda: Dietary, Lifestyle, and

Herbal Considerations, Center for

Integrative Medicine, the Ohio State

University, Columbus, OH, USA†,

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Uttarasthana, Chapter19th

Page No.173–

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Chauhan, Chetan R Patel, (2013)(1);

13–17 Scholars Research Library,

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Pharmacognostical and Microbial

Screening of Achyranthus aspera

(Amaranthaceae),

http://scholarsresearchlibrary.com, last

retrieved on April14

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Krishnanand, Badrinarayan Shastri, (1991)

Rastantrasar and Siddha prayogsangrah,

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edition, pg no.685

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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:dr.gibyabraham@gmail.com; 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

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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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