Scientific Basis for Ayurvedic Therapies -19

Scientific Basis for

Ayurvedic Therapies 

edited by

Brahmasree Lakshmi Chandra Mishra

15.5 Ayurvedic Herbs as Antimutagens

Phytochemicals are secondary metabolic products produced by plants in response to

environmental stresses. Thousands of these phytochemicals have been identified and,

when consumed in human diet, may affect chronic disease risk.

29

Of well over 2000

preparations known to the modern practitioner of Ayurveda, nearly 1500 are of plant

origin.

Susruta Samhita

refers to 700 drugs including a small number which were not

available in the country in that time. The list has grown substantially since then. Ancient

Indian medical literature has references not only to plants that cure difficult and incurable

diseases, but also to some endowed with many magical properties. Times have changed

and people are looking at logical causes and effects. India has been exposed for well over

a century to the application of allopathic medicines with their definite merits as well as

failings.

There is a return to some kind of natural healing and to Ayurveda and Siddha medicines.

The blind superstitious belief of the past has prompted extensive testing of several ancient

medicines. Out of the known 1500, well over 100 medicines have qualified for entry in

the Indian and British pharmaceutical Codex and the U.S. dispensatory. The investigations

have involved clinical and pharmacological testing of principal components, such as

phenolics and alkaloids, extracted from the herbs. Studies carried out on antimutagenic

effects of the Ayurvedic therapies were critically reviewed, their protocol studied, and the

significant results have been pointed out, as this effect may account for their therapeutic

effect to great extent. The Ayurvedic herbs discussed here with respect to their antimutagenic

and antiviral activity occupy an important place in the Ayurvedic system of

medicine and are used in the treatment of various ailments either alone or from a part of

various formulations.

15.6 Scientific Basis of Antimutagenic and Antiviral Activity of Ayurvedic

Therapies

Antimutagenic and antiviral activities of the herbs used in Ayurveda are presented. The

review indicates that the use of these herbs may be protective from exposure to environmental

mutagens.

15.6.1

Terminalia arjuna

With a view to explore Ayurvedic plants for chemopreventive activities,

T. arjuna

was

selected for testing of antimutagenic activity in our laboratory.

T. arjuna

is an important

cardiotonic plant described in Ayurveda and is widely used in the preparation of important

Ayurvedic formulations like

Arjunaristam, Cintamanirasam, Laksagulgulu,

Liv.52,

etc.

15.6.1.1 Antimutagenic and Antitumor Activity

Ellagic acid was isolated from

T. arjuna

and its antimutagenic potential was evaluated in

TA98 and TA100 strains of

Salmonella typhimurium

against direct- and indirect-acting

mutagens. It was found to be quite effective against promutagen 2-aminofluorene.

30

Various

other fractions were also isolated and were tested in Ames assay, comet, and micronucleus

(MN) tests, which indicate that the bark harbors constituents with promising

antimutagenic and anticarcinogenic potential.

31–33

Gallic acid, ethyl gallate, and luteolin

were reported to be active cancer-cell growth inhibitory constituents from

T. arjuna

by

Pettit and co-workers

34

using bioassay-guided separation methods. Kandil and Nassar

35

have reported an ellagitannin from the leaves of

T. arjuna

to be an anticancer promoter.

The effects of acetone and methanol extracts of

T. arjuna

were investigated on the growth

of human normal fibroblasts (WI-38), osteosarcoma (U2OS), and glioblastoma (U251) cells

in vitro

.

36

It was found that both extracts inhibited the growth of human U251 and U2OS

at 30 to 60

m

g/ml. It was also discovered that the extracts contained components that can

induce growth arrest of transformed cells by p53-dependent and independent pathways.

15.6.1.2 Antiviral Activity

Casuarinin, a hydrolysable tannin isolated from the bark, has been shown to possess antiherpes

virus activity.

37

15.6.2

Ocimum sanctum

Different parts of

O. sanctum

are traditionally used in Ayurveda and Siddha systems for

treating diverse ailments. Examples include infections, skin diseases, hepatic disorders,

common cold and cough, and malarial fever.

O. sanctum

is also used as an antidote for

snake bite and scorpion sting.

38

15.6.2.1 Antimutagenic and Antitumor Activity

O. sanctum

was one of the nine plant products that significantly decreased the incidence

of both benzo[a]pyrene (B[a]P)-induced neoplasia in the stomachs of Swiss mice and 3'-

methyl-4-dimethylaminoazobenzene (3'MeDAB)-induced hepatomas in Wistar rats.

39

The

topical treatment of the ethanolic leaf extract of

O. sanctum

significantly reduced the values

of tumor incidence, the average number of tumors per tumor bearing mice, and the

cumulative number of papillomas in 7,12-dimethylbenz[a]anthracene (DMBA)-induced

skin papillomagenesis in male Swiss albino mice at the periinitiational, postinitiational,

or continuously at both the peri- and postinitiational stages of papillomagenesis as compared

with the corresponding control group.

40

There was also significant twofold elevation

of reduced glutathione content in the skin of mice (

p

< 0.05) and an elevation of glutathione

S-transferase activity by 25% compared with the control group (

p

< 0.05) after 15 days of

the treatment.

In a study carried out by Banerjee et al.,

41

oral treatment with the alcoholic leaf extract

at 400 and 800 mg/kg body weight given to mice for 15 days significantly elevated the

activities of cytochrome P450 (

p

< 0.05), cytochrome b5 (

p

< 0.01,

p

< 0.001), aryl

hydrocarbon hydroxylase (

p

< 0.05), and glutathione S-transferase (

p

< 0.05,

p

< 0.01),

all of which are important in the detoxification of carcinogens as well as mutagens. The

reduced glutathione level in liver, lung, and stomach tissues was also elevated (

p

< 0.01,

p

< 0.001) by treatment with the leaf extract.

O. sanctum

leaf extract was also reported by Prashar and coworkers

42

to block or suppress

the events associated with chemical carcinogenesis by inhibiting metabolic activation of

the carcinogen. They treated the primary cultures of rat hepatocytes with up to 500

m

g of

O. sanctum

extract for 24 h and then with DMBA (10 or 50

m

g) for 18 h. A significant

reduction in the levels of DMBA-DNA adducts was observed in all cultures pretreated

with

O. sanctum

extract.

O. sanctum

also showed chemopreventive activity against DMBAinduced

hamster buccal pouch carcinogenesis.

43

O. sanctum

in the form of fresh leaf paste,

aqueous extract, and ethanolic extract were topically applied and the extracts were orally

administered to buccal pouch mucosa of animals exposed to 0.5% of DMBA. There was

significant reduction in the incidence of papillomas and squamous cell carcinomas. In

addition, there was an increase in the survival rate in the topically applied leaf paste and

orally administered extracts to animals. From the observations, it was suggested that the

orally administered extract of

O. sanctum

may have the ability to prevent the early events

of carcinogenesis.

Antiproliferative activity of seed oil of

O. sanctum

against HeLa cells in culture has also

been reported.

44

Prakash and Gupta

45

tested the chemopreventive activity of the seed oil

of

O. sanctum

against subcutaneously injected 20-methylcholanthrene-induced fibrosarcoma

tumors in the thigh region of Swiss albino mice. Supplementation of maximal

tolerated dose of 100

m

l/kg body weight of the oil significantly reduced the 20-methylcholanthrene-

induced tumor incidence and tumor volume. In liver enzymatic and nonenzymatic

antioxidants and lipid peroxidation end product, malondialdehyde levels were

significantly modulated with oil treatment as compared with untreated 20-methylcholanthrene-

injected mice. The potential chemopreventive activity of the oil was partly attributed

to its antioxidant properties.

15.6.2.2 Antiviral Activity

O. sanctum

is an ingredient of

tefroli,

which is used in condition of viral hepatitis. Rajalakshmi

and co-workers

46

have reported

O. sanctum

to show highly significant clinical and

biochemical clearance of viral hepatitis (

manjal kamalai

) when used as a single drug within

2 weeks of treatment. Patients for the above study were screened and selected from the

outpatient department of the Central Research Institute (Siddha) [C.R.I], Chennai, India,

based on the symptoms such as fever, dull ache in the right costal margin, yellow tint of

the sclera, anorexia, nausea and vomiting, dark-colored urine, clay-colored stools, and

enlarged and tender liver. Clinical diagnosis was confirmed by laboratory parameters such

as bile salts, bile pigment level in urine, and liver function test along with additional

laboratory parameters. Serum alkaline phosphatase, serum amylase, choleocystogram,

and stool tests were done to rule out the cases of surgical jaundice, neoplasm in the

abdominal cavity, obstruction in the biliary tract due to stone, and infestation with worms,

respectively. Twenty cases were screened and admitted in the inpatient department of CRI

for trial.

All the cases were assessed once a week with both clinical and biochemical parameters.

The leaves of

O. sanctum

were ground into paste and given in the dose of 10 g/day in

two divided doses. A diet of fat, tamarind, and spices with plenty of glucose was given

throughout the period of treatment. Within a week, anorexia, nausea, and vomiting disappeared

and stool regained its normal color. After 2 weeks of treatment, 50% of the cases

showed clearance of conjunctiva and dark-colored urine was seen only in 15% cases. The

reduction in the icteric index level was observed to be slow when compared with reduction

in serum bilirubin level. After 3 weeks of treatment, all the symptoms had cleared except

liver enlargement, which was persistent in 10% of the cases.

15.6.3

Glycyrrhiza glabra

The root of

G. glabra

is extensively used in traditional medicine and in food products as

a sweetening and flavoring agent.

15.6.3.1 Antimutagenic and Antitumor Activity

Zani et al.

47

investigated the effects of

G. glabra

extract, glycyrrhizinic acid, and 18

a

- and

18

b

-glycyrrhetinic acids on the mutagenicity of ethylmethanesulfonate (3

m

l/plate),

N

-

methyl-

N

'-nitro-

N

-nitrosoguanidine (1

m

l/plate), and ribose-lysine (25

m

l/plate) Maillard

model systems in TA100 tester strain of

S. typhimurium with the S. typhimurium and

microsome reversion assay. The compounds tested were also found to exhibit desmutagenic

activity, where the compound exerted its effect by direct interaction with the

mutagen, and antimutagenic activity, where the compound acted at the cellular level

suppressing the process of mutagenesis. All the compounds showed des-mutagenic activity

only against ribose-lysine mutagenic browning mixture. G. glabra extract showed

antimutagenic activity against ethylmethanesulfonate and ribose-lysine.

Glycyrrhizin, the main water-soluble constituent of licorice, was shown to possess

considerable antitumorigenic activity in Sencar mice. In this study, Agarwal et al.48 showed

that oral feeding of glycyrrhizin to Sencar mice resulted in substantial protection against

skin tumorigenesis caused by DMBA initiation and 12-O-tetradecanoylphorbol-13-acetate

(TPA) promotion. The latent period prior to the onset of tumor development was considerably

prolonged in glycyrrhizin-fed animals compared with animals not fed by glycyrrhizin.

Results showed a significant decrease in the number of tumors per mouse both

during and at the termination of the experiment. Oral feeding of glycyrrhizin in drinking

water also resulted in inhibition in the binding of topically applied [3H]B[a]P and

[3H]DMBA to epidermal DNA. The possible mechanism(s) of the antitumor-initiating

activity was attributed to the possible involvement of glycyrrhizin as an inhibitor of the

carcinogen metabolism followed by DNA adduct formation.

15.6.3.2 Antiviral Activity

Pompei et al.49 studied the effect of glycyrrhizic acid on the growth of vaccinia, herpes

simplex type 1 (HSV-1), Newcastle disease, vesicular stomatitis, and polio type 1 viruses

in cultures of human aneuploid HEp2 cells. Twenty-four-hour-old cell monolayers (107

cells per sample) were infected with five infectious units per cell of each virus at 20°C for

1 h, washed three times in Hank’s balanced saline solution (BSS), and incubated at 37°C

for 18 h in Eagle’s essential medium supplemented with 2% calf serum (pH 7.4). Infectious

virus yield was determined by the Dulbecco and Vogt technique and slightly modified

for vaccinia and HSV-1 viruses. Cytopathic effects were evidenced by observing Giemsastained

cells with a light microscope and by measuring spectrophotometrically at 530 nm

the amount of neutral red incorporated by cell cultures (100 mg/ml, 1-h pulses in drugfree

medium) after solubilization in 1% sodium deoxycholate in BSS. It was reported that

addition of 8mM glycyrrhizic acid after incubation completely inhibited both growth and

cytopathic effects of viruses except poliovirus type 1. It produced irreversible inactivation

of HSV-1 virus as the suspensions of this virus suffered a loss of infectivity of 105 when

incubated at 37°C with 8 mM glycyrrhizic acid for only 15 min. It was hypothesized that

glycyrrhizic acid interacted with sensitive virus proteins both at virionic stage and later,

when these are synthesized in host cells.

Glycyrrhizin was reported to inhibit varicella-zoster virus (VZV) in human embryonic

fibroblast (HEF) cells in vitro.50 On treatment of HEF with glycyrrhizin after inoculation

of virus (posttreatment), the average 50% inhibitory dose (ID50) for five VZV strains was

0.71 mM, and the selectivity index (ratio of ID50 for host-cell DNA synthesis to ID50 for

VZV replication) was 30. Glycyrrhizin was also effective against VZV replication when

HEF cells were treated 24 h before the inoculation (pretreatment). Furthermore, at a

concentration of 2.4 mM glycyrrhizin inactivated more than 99% of virus particles within

30 min at 37°C. Glycyrrhizin was reported to show a dose-dependent inhibition of the

replication of human immunodeficiency virus type 1 in MOLT-4 (clone No. 8) cells within

the concentration range of 0.075 to 0.6 mM.51 Within this concentration range, glycyrrhizin

also effected a dose-dependent reduction in the protein kinase C (PKC) activity

of MOLT-4 cells. PKC inhibition was considered as one of the mechanisms by which

glycyrrhizin inhibited human immunodeficiency virus type-1 (HIV-1) replication as a

PKC inhibitor, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride, also

proved inhibitory to HIV-1 replication in MOLT-4 cells.

Badam52 reported that indigenously purified glycyrrhizin was a more potent antiviral

agent than licorice from G. glabra and ammonium salt of glycyrrhizic acid (Sigma) in

inhibiting Japanese encephalitis virus (JEV) in vitro. Glycyrrhizin was found to inhibit

plaque formation in all the three strains of JEV — Nakayama, P-20778, and 821564 XY48

— at a concentration of 500 mg/ml at 96 h, whereas licorice and ammonium salt of

glycyrrhizic acid inhibited at 1000 mg/ml concentration. A daily injection of glycyrrhizin

(stronger neo-minophagen C [SNMC] containing 40 mg glycyrrhizin in a 20-ml ampoule)

was reported to lower alanine aminotransferase (ALT) levels in patients with chronic viral

hepatitis.53 The therapeutic effects of intermittent administration of SNMC three times/

week for 12 weeks were evaluated and compared between two doses (40 and 100 ml) in

a randomized clinical trial. The therapeutic response was better in the 53 patients allocated

100 ml than in the 59 who were allocated to have 40 ml of SNMC. At the completion of

SNMC treatment, ALT levels decreased more extensively in the patients on 100 ml than

in those on 40 ml of SNMC. Minor side effects occurred in both the patients on 100 ml of

(20%) and in those on 40 ml (12%) but did not require any therapies. It was suggested

that intermittent SNMC would be efficient in suppressing ALT levels in patients with

chronic viral hepatitis in a dose-dependent manner.

15.6.4 Semecarpus anacardium

S. anacardium Linn. of the family Anacardiaceae has many applications in the Ayurvedic

and Siddha systems of medicine. It is popularly known as multipurpose medicine (ardha

vaidya).

15.6.4.1 Antimutagenic and Antitumor Activity

Water, alcohol, and oil extracts of S. anacardium were found to be antimutagenic against

B[a]P-induced mutagenicity in TA98 and TA100 tester strains of S. typhimurium.54 Methanol

extract, resinous fraction, and Bhilawanol isolated from S. anacardium showed antitumour

activity against P388 lymphocytic leukemia in BDF1 mice as judged by their median

survival time.55 Smit and co-workers56 studied 14 specimens from the list of Ayurvedic

herbal drugs and collected from various parts of India and Nepal for cytostatic activity.

© 2004 by CRC Press LLC

Antimutagenic Effect of Ayurvedic Therapies 263

The ethanolic (70% v/v) extracts were tested for cytotoxicity on COLO 320 tumor cells,

using the microculture tetrazolium assay. The nuts of S. anacardium displayed a cytotoxic

effect with a IC50-value of 1.6 mg/ml.

Premalatha et al.57 have reported the modulating effect of the extract of SA against

aflatoxin B1-induced experimental hepatocellular carcinoma. Anticancer property was

attributed to its strong antioxidant capacity and its capability to induce in vivo antioxidant

system.58 In a study,59 the beneficial effect of SA in the treatment of hepatocellular carcinoma

was attributed to the stabilization of biomembranes by S. anacardium nut extract. S.

anacardium nut milk extract was administered orally at a dose of 200 mg/kg/day for 14

days to normal rats as well to animals whose biomembranes were rendered fragile by the

induction of hepatocellular carcinoma with aflatoxin B1, where the discharge of lysosomal

enzymes increased significantly with a subsequent increase in glycoprotein components.

The nut extract administration reversed these adverse changes to near normal in treated

animals. The administration of the same dose to male albino rats with aflatoxin B1-induced

hepatocellular carcinoma was found to be highly effective in inducing phase I and phase

II biotransformation enzymes.60

The administration of S. anacardium nut extract has also been reported to cause a significant

decrease in the activity of glycolytic enzymes and an increase in gluconeogenic

enzymes activities to near normal values in drug-treated animals.61 Anacartin forte, an

Ayurvedic preparation, exhibited not only a broad spectrum of anticancer properties in

clinical and animal studies, but also a wide margin of safety in therapeutic dosage even

when used for long periods. It showed satisfactory results in cases of cancer of the

esophagus, liver, urinary bladder, liver, and chronic leukemia by giving subjective and

objective improvement, alleviation, or disappearance of troublesome symptoms and clinical

benefit with extension of survival time. The preparation has selective action, attacking

only the cancer cells without harming the normal cells.62

15.6.5 Terminalia chebula

T. chebula is a rasayana to vata, increasing awareness, and has a tonic effect on the central

nervous system. It improves digestion, promotes the absorption of nutrients, and regulates

colon function. T. chebula is most useful in prolapsed organs, improving the strength and

tone of the supporting musculature.

15.6.5.1 Antimutagenic and Antitumor Activity

Water extract of T. chebula was shown to significantly reduce 4-nitro-o-phenylenediamine

(NPD) as well as 2-AF induced his+ revertants in Ames assay.63 A tannin fraction (TC-E)

obtained from the dried fruit pulp of T. chebula was subjected to column chromatography

to yield four fractions — TC-EI, TC-EII, TC-EIII, and E-IV — which were evaluated for

their antimutagenic potential. The fractions were quite effective in inhibiting the mutagenicity

of 2-AF. The monomeric fraction (TC-EI) was the least effective in comparison with

other oligomeric fractions.64 A 70% methanol extract of T. chebula fruit was studied for its

effects on growth in several malignant cell lines including a human (MCF-7) and mouse

(S115) breast cancer cell line, a human osteosarcoma cell line (HOS-1), a human prostate

cancer cell line (PC-3), and a nontumorigenic immortalized human prostate cell line

(PNT1A) by using assays for proliferation, cell viability, and cell death. In all cell lines

studied, the extract decreased cell viability, inhibited cell proliferation, and induced cell

death in a dose-dependent manner.65

   

15.6.5.2 Antiviral Activity

Badmaev and Nowakowski66 tested a multicomponent herbal formula, ledretan-96, consisting

of 23 components on an epithelial tissue culture cell line (MDCK) for its protective

activity against cytopathic effects caused by influenza A virus. The whole formula and

each of its 23 individual components were tested in the same system. The results indicated

that the formula, when prepared according to established procedure, in the form of

decoction was active in protecting epithelial cells against damage caused by influenza A

virus used at different dosages. Of the 23 components tested, only T. chebula showed a

significant protective effect when applied to the epithelial cells individually. The bioassaydirected

isolation of T. chebula fruits afforded four HIV-1 integrase inhibitors, gallic acid,

and three galloyl glucoses.67 The galloyl moiety played a major role for inhibition against

the 3'-processing of HIV-1 integrase.

T. chebula has also been reported to show a significant inhibitory activity on human

immunodeficiency virus reverse transcriptase.68 Anti-HSV-1 activity of T. chebula has been

reported by Kurokawa et al.69 T. chebula reduced virus yield in the brain and skin more

strongly than acyclovir alone and exhibited stronger anti-HSV-1 activity in the brain than

in the skin, in contrast to acyclovir treatment by itself. A group70 from Japan showed T.

chebula to possess anticytomegalovirus (CMV) activity. Shiraki et al.71 reported T. chebula

as one of the medicinal plants to inhibit replication of human CMV and murine CMV in

vitro and suggested it to be beneficial for the prophylaxis of CMV diseases in immunocompromized

patients.

15.6.6 Terminalia bellerica

T. bellerica is also a strong rejuvenator of the body and is recommended as a daily supplement.

15.6.6.1 Antimutagenic Activity

Two polyphenolic fractions isolated from T. bellerica were significantly effective against

mutagenic effects in S. typhimurium. Interaction of the polyphenols with S9 proteins may

be the probable cause of the inhibitory effect.72

15.6.6.2 Antiviral Activity

Suthienkul et al.73 reported the extract of T. bellerica to show retroviral reverse transcriptase

inhibitory activity. Hot water and methanol extracts of 57 Thai herbs and spices were

examined for their retroviral reverse transcriptase inhibitory activity with Moloney murine

leukemia virus reverse transcriptase. Hot water extract of T. bellerica at a concentration of

125 mg/ml showed relative inhibitory ratio of 75%, whereas its methanol extract exhibited

an inhibitory ratio (IR) value of 83%. An extract of T. bellerica showed significant inhibitory

activity on HIV-1 reverse transcriptase, with IC50 £50 mg/ml.68 Four lignans isolated possessed

demonstrable anti-HIV-1 activity in vitro.74

15.6.7 Emblica officinalis

E. officinalis is an important traditional medicine with broad prospects. The fruits of the

plant have been used in Ayurveda as potent rasayanas and form the major constituent of

chyavanprash awaleha and triphala.

15.6.7.1 Antimutagenic and Antitumor Activity

Water, acetone, and chloroform extracts of E. officinalis fruit were reported to significantly

reduce the mutagenicity of sodium azide and NPD in TA100 and TA97a strains, respectively,

of S. typhimurium.75 E. officinalis extract was reported to show a pronounced protective

effect in counteracting the genotoxicity induced by aluminium and lead.76,77 Oral

administration of E. officinalis extract for 7 consecutive days before the exposure of mice

to the metals by intraperitoneal injections reduced the frequencies of sister chromatid

exchanges and micronuclei-induced in bone marrow cells by both metals. Aqueous extract

of E. officinalis was reported to be quite effective in inhibiting mutagenicity of S9-dependent

mutagens, aflatoxin B1 (0.5 mg/plate), and B[a]P (1 mg/plate) in TA98 and TA100 tester

strains of S. typhimurium in Ames assay.78

Dietary supplementation with extract of fruit of E. officinalis to Swiss albino mice significantly

reduced the cytotoxic effects of 3,4-B[a]P in vivo.79 Age-matched Swiss albino

mice were fed by gavaging the fruit extract daily for 28 days, and one dose of the

carcinogen was given on alternate days up to a total of eight doses from day 9. On day

29, all mice were transferred to normal diet. Control sets received the extract alone, the

carcinogen alone, and olive oil alone. All mice were sacrificed at 12 weeks and 14 weeks

after the end of the experiment. Chromosome preparations were made from bone marrow

after the usual colchicine-hypotonic-fixative-airdrying-Giemsa staining schedule. The end

points screened were the frequencies of chromosomal aberrations and damaged cells that

were induced, which clearly pointed toward the modulation of B[a]P-induced cytotoxic

effects by the fruit extract.

Sharma and co-workers80 studied the effect of E. officinalis extract administration on the

in vivo genotoxicity of B[a]P and cyclophosphamide (CP) using bone marrow chromosomal

aberration and micronucleus induction tests in mice. Three doses (50, 250, and 500 mg/

kg body weight) of the plant extract were administered orally for 7 consecutive days prior

to the administration of single dose of mutagens (B[a]P 125 mg/kg oral; CP 40 mg/kg

intraperitoneal). It was found that administration of 250 and 500 mg/kg of E. officinalis

extract significantly inhibited the genotoxicity of B[a]P as well as CP in both the assay

systems. There was a significant induction in the levels of glutathione content (GSH) and

of antioxidant and detoxification enzymes. Extract of E. officinalis significantly inhibited

hepatocarcinogenesis induced by N-nitrosodiethylamine (NDEA) in a dose-dependent

manner.81 Its anticarcinogenic activity was evaluated by its effect on tumour incidence,

levels of carcinogen metabolizing enzymes, levels of liver cancer markers, and liver injury

markers, which clearly indicated its protection against chemical carcinogenesis.

In a study,82 in vitro antiproliferative activity of extracts from medicinal plants were

compared with human tumor cell lines, including human erythromyeloid K562, B-lymphoid

Raji, T-lymphoid Jurkat, and erythroleukemic HeLa cell lines. Extracts from E.

officinalis were the most active in inhibiting in vitro cell proliferation and contained pyrogallol

as an active component. Aqueous extract of E. officinalis has also been reported by

Jose et al.83 to be cytotoxic to L929 cells in culture in a dose-dependent manner. The

concentration needed for 50% inhibition was found to be 16.5 mg/ml. E. officinalis and

chyavanaprash extracts were also found to reduce ascites and solid tumors in mice induced

by Dalton's lymphoma ascites (DLA) cells. Animals treated with 1.25 g/kg body weight

of E. officinalis extract increased the life span of tumor-bearing animals by 20%, whereas

animals treated with 2.5 g/kg body weight of chyavanaprash produced a 60.9% increase

in the life span. Antitumor activity of E. officinalis extract was attributed partially to its

interaction with cell cycle regulation as was found to inhibit cell cycle-regulating enzymes.

15.6.7.2 Antiviral Activity

A bioassay-guided fractionation of a methanol extract of the fruit of E. officinalis yielded

Putranjivain A (1) as a potent inhibitory substance on the effects of HIV-1 reverse transcriptase

with IC50 = 3.9 mM, together with (2) 1,6-di-O-galloyl-b-D-glucose, (3) 1-O-galloylb-

D-glucose, (4) kaempferol-3-O-b-D-glucoside, (5) quercetin-3-O-b-D-glucoside, and (6)

digallic acid. The inhibitory mode of action by 1, 2, and 6 was noncompetitive with respect

to the substrate but competitive with respect to a template-primer.68

15.6.8 Cinnamomum cassia

C. cassia is known as dalchini in the Indian subcontinent. The bark of C. cassia is frequently

used in Ayurveda and Unani preparations.

15.6.8.1 Antimutagenic Activity

Sharma et al.84 evaluated the antimutagenic effect of C. cassia against two mutagens, B[a]P

and CP, by in vivo chromosomal aberration and micronuclei tests after pretreatment with

the extract orally for 7 consecutive days and with Ames assay. C. cassia pretreatment

decreased cytochrome P450 content but increased GSH content and the activity of glutathione-

dependent antioxidant enzymes. These results indicate its modulatory effect on

the xenobiotic bioactivation and detoxification processes. 2'-Hydroxycinnamaldehyde isolated

from C. cassia was found to strongly inhibit in vitro growth of 29 kinds of human

cancer cells and in vivo growth of SW-620 human tumor xenograft without the loss of

body weight in nude mice.85 It prevented adherence of SW-620 cells to the culture surface

but did not inhibit oncogenic K-Ras processing, implying its antitumor mechanisms at the

cellular level. Kwon et al.86 found the key functional group of the cinnamaldehyde-related

compounds in the antitumor activity to be in the propenal group.

15.6.8.2 Antiviral Activity

C. cassia was one of the many medicinal plants possessing potent anti-HIV activity. Premanathan

et al.87 studied the inhibitory effect of plant extracts on HIV replication in terms

of the inhibition of virus-induced cytopathogenicity in MT-4 cells. The MT-4 cells were

infected with HIV. The HIV-infected MT-4 cells were incubated at 37°C in a CO2 incubator

in the presence of the plant extracts. After 5 days, cell viability was measured by a

tetrazolium-based colorimetric assay.

15.6.9 Withania somnifera

W. somnifera is one of the Indian medicinal plants having a remarkable reputation, as a

factor of health care, among the indigenous medical practitioners. Several studies over

the past few years have indicated that W. somnifera has anti-inflammatory, antitumor,

antistress, antioxidant, mind-boosting, and rejuvenating properties.

15.6.9.1 Antitumor Activity

The alcoholic extract of the dried roots of the plant, as well as the active component

withaferin A isolated from the extract, showed significant antitumor and radiosensitizing

effects in experimental tumors in vivo, without any noticeable systemic toxicity.88 Russo

et al.89reported the effect of methanolic extract of W. somnifera in reducing the hydrogen

peroxide-induced cytotoxicity and DNA damage in human nonimmortalized fibroblasts.

The extract showed dose-dependent free radical scavenging capacity and a protective

effect on DNA cleavage.

The chemopreventive activity of a hydroalcoholic extract of W. somnifera roots against

20-methylcholanthrene induced fibrosarcoma tumors in Swiss albino mice was reported

by Prakash et al.90 A single subcutaneous injection of 200 mg 20-methylcholanthrene in 0.1

ml of dimethyl-sulphoxide into the thigh region of mice produced a high incidence (96%)

of tumors. Oral treatment of animals with 400 mg/kg body weight of extract (1 week

before injecting 20-methylcholanthrene and continuing until 15 weeks thereafter) significantly

reduced the tumor incidence and tumor volume and enhanced the survival of the

mice, compared with 20-methylcholanthrene–injected mice. Liver biochemical parameters

revealed a significant modulation of reduced glutathione, lipid peroxides, glutathione-Stransferase,

catalase, and superoxide dismutase in extract-treated mice. The extract was

also quite effective in preventing DMBA-induced squamous cell carcinoma of skin in Swiss

albino mice.91 The skin lesions were induced by the twice-weekly topical application of

DMBA (100 nmol/100 ml acetone) for 8 weeks on the shaved backs of mice. The extract

was administered at the maximal tolerated dose of 400 mg/kg body weight three times/

week on alternate days 1 week before DMBA and continued for 24 weeks thereafter. The

chemopreventive activity was also linked to the antioxidant and free radical-scavenging

constituents of the extract.

15.6.10 Centella asiatica

Centella asiatica (Umbelliferae) syn. Hydrocotyl asiatica has been used in various parts of

India for different ailments. Examples include headaches, body aches, insanity, asthma,

leprosy, ulcers, eczemas, and wound healing.

15.6.10.1 Antimutagenic and Antitumor Activity

The researchers at the Amala Cancer Research Centre in Kerala, India, tested both crude

extract of C. asiatica and its partially purified fractions (AF) for their antitumor activity.

AF inhibited the proliferation of the transformed cell lines significantly more than the

crude extract and other solvent fractions in dose-dependent manner. Fifty percent effective

doses of AF on its 3-h exposure for Ehrlich ascites tumor cells (EAC) and DLA were

reported to be 17 and 22 mg/ml, respectively. AF also significiantly suppressed the multiplication

of mouse lung fibroblast (L-929) cells at a concentration of 8 mg/ml in longterm

culture. Oral administration of the extracts retarded the development of solid and

ascites tumors and increased the life span of these tumor-bearing mice. Tritiated thymidine,

uridine, and leucine incorporation assay suggested that the fraction acted directly

on DNA synthesis.92 Yen et al.93 reported the inhibitory effect of C. asiatica against the

mutagenicity of 2-amino-3-methyl-imidazole (4,5-f) quinoline.

15.6.10.2 Antiviral Activity

Aqueous extract of C. asiatica was one of the 500 herbs tested that showed significant anti-

HSV-II action as determined by the virus inhibition logarithm.94 Yoosook et al.95 reported

asiaticoside from C. asiatica to be an active constituent against antiherpes simplex virus.

It showed both anti-HSV-1 and 2 activities in plaque inhibition assay.

15.7 Conclusion

A search of the literature on Ayurvedic herbs revealed ten herbs that have studies showing

antimutagenic and antiviral activity: T. arjuna, O. sanctum, G. glabra, S. anacardium, T.

chebula, T. bellerica, E. officinalis, C. cassia, W. somnifera, and C. asiatica. The data are indicative

of their possible protective effect against the environmental mutagens. Future research is

needed to further confirm their antimutagenic and antiviral effects in animals and humans.

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Benign Growths, Cysts, and Malignant Tumors

Manoranjan Sahu and Lakshmi Chandra Mishra

Om Tat Sat

                                                        

(Continued...) 

(My humble salutations to H H Maharshi ji,  Brahmasri Sreeman Lakshmi Chandra Mishra ji and other eminent medical scholars and doctors   for the collection)