C:^JlTTT7l III- 7i:tyi:t}V Of£I7!E2lA7tlJi:F

"Reyiew of Citerature

3.1. Review on Curcuma longa

The turmeric (Curcuma longa) plant, a perennial herb belonging to the Zingiberaceae family, is

cultivated extensively in south and southeast tropical Asia. The rhizome of this plant is also

referred to as the "root" and is the most useful part of the plant for culinary and medicinal

purposes. The most active component of turmeric is curcumin, which makes from 2 to 5% of the

spice. The characteristic yellow color of turmeric is due to the curcuminoids, first isolated by

Vogel in 1842. Turmeric contains (Kapoor r̂. al., 1990).

Protein

6.30%

Fat

5.10%

Minerals

3.50%

Carbohydrate

69.40%

moisture

13.10%

The essential oil (5.8%) obtained by steam distillation of rhizomes has (Kapoor et. al., 1990)

a-phellandrene

(1%)

sabinene

(0.6%)

cineol

(1%)

borneol

(0.5%)

zingiberene

(25%)

sesquiterpines

(53%)

Curcumin (diferuloylmethane) (3-4%) is responsible for the yellow colour, and comprises

curcumin I (94%), curcumin II (6%) and curcumin III (0.3%) (Ruby et. al., 1995). Demethoxy

and bisdemethoxy derivatives of curcumin have also been isolated (Vopel et. al., 1990).

Curcumin was first isolated in 1815 and its chemical structure was determined by Roughley and

Whiting,in 1973. It has a melting point at 176-177°C; forms a reddish-brown salt with alkali and

is soluble in ethanol, alkali, ketone, acetic acid and chloroform (Chattopadhyay et. al., 2004).

C. longa is reported to inhibit the growth of some bacteria and fungi (Apisariyakul et. al., 1995;

Singh et. al., 2002; Chauhan et. ai, 2003). It had antibacterial activities against MRSA and

lowered the MIC's of p-lactams (Kang et. al., 2005). Mahady et. al., (2002) revealed that both

11

'Re'view of Citerature

the methanol extract of turmeric and curcumin inhibited the growth of H. pylori infection. The

water extract of Curcuma longa inhibit the growth of H. pylori (Behrooz et. al, 2008). Turmeric

fractions inhibited the growth of some intestinal and pathogenic bacteria in vitro (Shankar,

1979). Curcumin has also been shown to exhibit antimicrobial activities (Anand et. al., 2007).

Negi et. al., (1999) demonstrated that turmerone and curlone components of turmeric oil possess

excellent antibacterial action against a wide range of microbes, such as Bacillus cereus, Bacillus

coagulans, Bacillus subtills, Staphylococcus aureus, Escherichia coli, and Pseudomonas

aeruginosa. Turmeric also induce predominant anti-fungal activity as reported by

Wuthiudomelert et. al, (2000). Behura et. al, (2000) reported that essential oil of C. longa leaf

has anti-fungal action as similar to standard fungicides like cabendazin and manocozeb.

Venugopal and Saju et. al., (1999) reported that turmeric oil exhibits excellent insect repellent

property even at 1 % concentration in water.

Curcumin is also well known for its therapeutic properties. The modulatory effect of dietary

curcumin (0.05%, w/w) on drug metabolizing and general marker enzymes of liver and

formation of Aflatoxin Bi-adducts (DNA and protein adducts) due to dietary Aflatoxin

Bi exposure for a period of 6 weeks have been evaluated in a rodent model (Nayak and

Sashidhare/. al, 2010).

Curcumin is a potent antioxidant and anti-inflammatory agent with hepatoprotective, anti-

carcinogenic and antimicrobial properties (Pal et. al., 2001). In in vivo evaluation of curcumin

soya-lecithin complex in paracetamol-induced hepatotoxicity in mice, it was observed that the

complexed curcumin afforded a significantly higher protection against paracetamol-induced rise

in serum aspartate aminotransferases and alanine aminotransferase and alanine transferase levels

as compared to pure curcumin (Mukesh et. al, 2008). Deshpande et. al., (1998) carried out a

12

'Review of Citerature

study on hepatoprotective activity in rats. They have reported that pretreatment with C. longa

caused reduction in bilirubin, cholesterol, AST and alkaline phosphatase activity in CCI4 induced

liver toxicity in animal model. Report by Rajsekharan et. ai, (1998) showed the protective effect

of curcumin in ethanol induced liver toxicity. Kandarkar et. ai, (1998) reported that dietary

administration of turmeric or ethanolic extract of turmeric for 14 days at cancer preventive doses

was found to be hepatotoxic in mice as observed by histopathologic and ultrastructural studies.

Soni et. ai, (1997) showed protective effect of food additives on aflatoxin induced mutagenicity

and hepatocarcinogenicity. Singh and Aggarwal (1995) reported postnatal modulation of hepatic

biotransformation system enzymes via translactational exposure of Fl mouse pups to turmeric

and curcumin. Curcumin significantly prevented the serum levels of AST and ALT in iron-

treated rats, indicating that this spice principle reduced the severity of iron toxicity by reducing

the lipid peroxidation (Reddy and Lokesh, 1994). Ethanol administration resulted in biochemical

and histopathological changes in the liver, kidney and brain that were reverted when curcumin

and N-acetylcysteine were given to rats intoxicated with ethanol (Rajakrishnan et. ai, 1999).

Curcumin has been shown to inhibit the production of arachidonic acid in the liver, kidney and

brain (Rajakrishnan et. ai, 2000). Rajakrishnan et. al., (1999) found a significant increase in the

levels of Prostaglandin PGEl, Prostaglandin PGE2, Prostaglandin PGF2a and Prostaglandin

PGD2 in the liver, kidney and brain of alcohol-fed rats, while co treatment with curcumin

decreased the level of prostaglandins significantly. Curcumin helps in maintaining the membrane

structure, integrity and function of vital organs, protecting the liver, brain and kidney from

alcohol toxicity (Rajakrishnan et. al., 2000). The findings of Shapiro et. ai, (2006) study confirm

a role of ROS, nitric oxide and NF-kB activation in the pathogenesis of thioacetamide-induced

liver damage and indicate that a blockade of their formation or activation by curcumin

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'Review of Citerature

administration has a potent anti-inflammatory effect. Because curcumin is safe in humans

(Chainani Wu, 2003), its use in patients with acute liver injury can be evaluated based on these

results. Curcumin protects against inflammation and oxidative stress in carbon tetrachloride

(CCl4)-induced acute hepatic injury (Reyes et. al., 2007). Studies involving systemic curcumin

administration have demonstrated its beneficial effects by modulating NF-kB activity (Singh et.

al., 1995); in addition, the ability of curcumin to scavenge a variety of ROX (Maheswari et. al.,

2006) makes this compound a suitable tool to be studied in CCU-induced liver damage. The

study of Park et. al., (2000) demonstrated that curcumin can effectively inhibit the hepatic

damage produced by either acute or subacute CCI4 treatment as monitored by serum biochemical

parameters, fibrosis and lipid peroxides in the liver. The ability of oral curcumin (200 mg/kg) in

preventing acute CCI4 (4 g/kg, p.o.) intoxication was studied recently (Reyes et. al, 2007).

Anti-inflammatory activity of turmeric oil has also been reported on pepper's model

(Ramachandran et.al, 2000). Mishra et. al, (1997) reported that the volatile oil of C. longa was

effective in anti-inflammatory and anti-hyaluronidase action. They suggested that it has

antioxidative effect as evidenced by inhibition of diffusion capability of the hyaluronidase

enzyme by the oil. Further the cytotoxic, anti-inflammatory and antioxidant activity of curcumin

I, II and III from C. longa was studied by Ramsewk et al., (2000). They observed cytotoxic

activity against leukemia, colon cancer, CNS melanoma, renal and breast cancer. Leaf oil of C.

longa also showed potent anti-inflammatory activity in carranngenin induced paw edema in male

albino rats as reported by Iyengar et. al., (1994). Anto et. al., (1996) reported that curcumin III is

a most potent anti-inflammatory agent amongst present natural curcuminoids (I, 11, III) and other

B-synthetic curcuminoids. Lantz et. al., (2005) reported the effect of turmeric extracts on

14

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inflammatory mediator production. Ammon et. al., (1993) suggested mechanism of anti­

inflammatory actions of curcumin. Kulkami et. al., (1991) proposed treatment of osteoarthritis

with a herbomineral formulation: a doubleblind, placebo-controlled, cross-over study.

Huang et. al., (1992) demonstrated the inhibitory effect of curcumin, an anti-inflammatory agent,

on vascular smooth muscle cell proliferation.

Turmeric was found to be effective in treatment of allergy. Studies on anti-allergic activity

(Yano et. al., 2000 and 1996) were carried out on various extract of Curcuma longa rhizome.

The ethyl acetate fraction was found to be most potent anti-allergic agent amongst other extracts.

This causes potent inhibition of histamine release from mast cells. Crude extract of fresh rhizome

of C. longa was found to posses good cycloxygenase (COX) inhibitory action (Venkatatshwarlu,

1997) in an in-vitro bioassay test. Turmeric is established as an excellent remedy for wound

healing since antiquity. The local application of Curcuma longa powder efficiently heals septic

wounds in diabetic patients (Pandya ,1995).

Numerous lines of evidence suggest that curcumin can modulate both the proliferation and the

activation of T cells (Ranjan et. al., 2004). Curcumin exhibits immunosuppressive effects

mediated through regulation of IL-2. Another study on mouse lymphocytes has reported that a

low-dose curcumin exposure increased the proliferation of splenic lymphocytes, whereas at high-

doses curcumin depressed it, indicating its ability to differentially regulate the proliferation of

splenic lymphocytes (Li and Liu, 2005). The experiments on rat splenic lymphocytes showed

that curcumin treatment enhances the immune response of the lymphocytes by increasing IgG

production (Kuramoto et. a/., 1996). Curcumin not only plays an important role in the

immunomodulation of normal but also transformed T cells, where it adversely affects the cell

15

'Review of Citerature

proliferation of these cells by suppression of IL-2 gene expression and by inhibiting the

activation of NF-ATB (Gertsch et. al, 2003). Curcumin treatment stimulates proliferation of B

cells in the mucosa of intestine of C57BL/6J-Min/+ (Min/+) mice indicating its

immunostimulatory activity (Churchill et. al., 2000). Studies have shown curcumin's ability to

modulate the activation of macrophages, e.g. regulation of the immune function of mice in a

dose-dependent fashion as curcumin treatment enhanced the phagocytosis of peritoneal

macrophages and differentially regulates the proliferation of splenocytes (Li and Liu, 2005).

Apart from cell proliferation, a daily diet of curcumin (30 mg/kg body weight/day) for 2 weeks

in rats reportedly attenuated the ability of macrophages to generate ROS, (Joe et. a/., 1994) and

secrete lysosomal enzymes collagenase, elastase, and hyaluronidase (Joe et. al., 2000). The

ability of curcumin to downregulate Thl and NO production has been directly correlated to its

ability to differentially activate the host macrophages (Bhaumik et. al., 2000). Curcumin can also

apparently modulate the activation of natural killer (NK) cells. Studies by South and his

colleagues, in rats showed that curcumin at a dose of 1 and 20 mg/kg body weight could not

enhance the IgG levels in the NK cells, whereas a higher dose (40 mg/kg) did elevate IgG levels

significantly. More importantly, none of the three doses of curcumin significantly enhanced

either delayed-type hypersensitivity or NK cell activity (South et. al., 1997). There is only one

report to date on immune modulation of murine dendritic cells (DCs) using curcumin by Kim et.

al, who found that curcumin significantly depressed the expression of CD80, CD86, and MHC

class II antigens in GM-CSF/IL-4 stimulated DCs without affecting MHC class I antigens. In yet

another study, curcumin caused cell death by apoptosis in both normal and transformed human

(HL 60) and rodent cells despite the lack of oligonucleosomal DNA fragmentation (DNA

"ladder"). However, curcumin blocked HL-60 in sub-Gl and increased caspase-3 activity

16

Jieviey^ of Citerature

(Bielak et. al., 2000). These results indicate that curcumin exerts its immunomodulatory action

on other immune cells. The effect of curcumin in Alzheimer's disease is mediated through the

downmodulation of cytokine (i.e., TNF-a and IL-]/]) and chemokine (i.e., MlP-lb, MCP- 1, and

IL-8) activity in peripheral blood monocytes and reduces amyloid-y5 plaque formation (Lim et.

al., 2001; Giri et. al, 2004; Yang et. al., 2005 and Zhang et. al., 2006).

Turmeric powder has beneficial effect on the stomach. It increases mucin secretion in rabbits and

may thus act as gastroprotectant against irritants (Lee et. al, 2003). However, controversy exists

regarding antiulcer activity of curcumin. Both antiulcer (Sinha et. al,1915) and ulcerogenic

(Prasad et. al, 1976; Gupta et. a/., 1980) effects of curcumin have been reported but detailed

studies are still lacking. Curcumin has been shown to protect the stomach from ulcerogenic

effects of phenylbutazone in guinea pigs at 50 mg/kg. It also protects from 5-hydroxytryptamine-

induced ulceration at 20 mg/kg dose in mice model (Dasgupta et. al, 1969; Sinha et. al, 1974).

In fact, at higher doses of 50 mg/ kg and 100 mg/kg, it produces ulcers in rats (Sinha et. al,

1975). Though the mechanism is not yet clear, an increase in the gastric acid and/or pepsin

secretion and reduction in mucin content have been implicated in the induction of gastric ulcer

(Cream et. a/., 1974). Curcumin decreases the severity of pathological changes and thus protects

from damage caused by myocardial infarction (Nirmala and Puvanakrishnan, 1996). Curcumin

improves Ca^^-transport and its slippage from the cardiac muscle sarcoplasmic reticulum,

thereby raising the possibility of pharmacological interventions to correct the defective Ca "̂

homeostasis in the cardiac muscle (Sumbilla et. al, 2002). Curcumin and manganese complex of

curcumin offer protective action against vascular dementia by exerting antioxidant activity

(Vajragupta et. al, 2003; Thiyagarajan and Sharma, 2004).

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'Review of Citerature

Curcumin reduces low density lipoprotein and very low density lipoprotein significantly in

plasma and total cholesterol level in liver along with an increase of a-tocopherol level in rat

plasma, suggesting in vivo interaction between curcumin and a-tocopherol that may increase the

bioavailability of vitamin E and decrease cholesterol levels (Kamal et. al, 2000). Curcumin

binds with egg and soy-phosphatidylcholine, which in turn binds divalent metal ions to offer

antioxidant activity (Began et. al, 1999). The increase in fatty acid content after ethanol-induced

liver damage is significantly decreased by curcumin treatment and arachidonic acid level is

increased (Akrishnan and Menon, 2001).

Curcumin acts as a scavenger of oxygen free radicals (Subramanian et. a/., 1994). In vitro,

curcumin can significantly inhibit the generation of reactive oxygen species (ROS) like

superoxide anions and H2O2 as well as nitrite radical generation by activated macrophages,

which play an important role in inflammation (Joe and Lxjkesh, 1994). Its derivatives,

demethoxycurcumin and bis-demethoxycurcumin also have antioxidant effect (Unnikrishnan and

Rao, 1995; Song et. al, 2001). Curcumin exerts powerful inhibitory effect against H202-induced

damage in human keratinocytes and fibroblastsS 1 and in NG 108-15 cells (Mahakunakom et. al,

2003). It also decreases lipid peroxidation in rat liver microsomes, erythrocyte membranes and

brain homogenates (Pulla and Lokesh, 1994). As this damage was prevented by antioxidant a-

tocopherol, the pro-oxidant role of curcumin has been proved. Curcumin also causes oxidative

damage of rat hepatocytes by oxidizing glutathione and of human erythrocyte by oxidizing

oxyhaemoglobin, thereby causing haemolysis (Galati et. al, 2002).

Petroleum ether and aqueous extracts of turmeric rhizomes show 100% antifertility effect in rats

when fed orally (Garg, 1974). Implantation is completely inhibited by these extracts (Garg et.

a/., 1978). Curcumin inhibits 5a-reductase, which converts testosterone to 5a-

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'Review of Citerature

dihydrotestosterone, thereby inhibiting the growth of flank organs in hamster (Liao et. ai, 2001).

Curcumin also inhibits human sperm motility and has the potential for the development of a

novel intravaginal contraceptive (Rithapom et. ai, 2003).

Curcumin prevents galactose-induced cataract formation at very low doses (Suryanarayana et.

ai, 2003). Both turmeric and curcumin decrease blood sugar level in alloxan-induced diabetes in

rat (Arun and Nalini, 2002). Curcumin also decreases advanced glycation end products induced

complications in diabetes mellitus (Sajithlal et. a/., 1998).

Curcumin exerts both pro- and antimutagenic effects. At 100 and 200 mg/kg body wt doses,

curcumin has been shown to reduce the number of aberrant cells in cyclophosphamide- induced

chromosomal aberration in Wistar rats (Shukla et. al, 2002).

Curcumin acts as a potent anticarcinogenic compound. Among various mechanisms, induction of

apoptosis plays an important role in its anticarcinogenic effect. It induces apoptosis and inhibits

cell-cycle progression, both of which are instrumental in preventing cancerous cell growth in rat

aortic smooth muscle cells. The antiproliferative effect is mediated partly through inhibition of

protein tyrosine kinase and c-myc mRNA expression and the apoptotic effect may partly be

mediated through inhibition of protein tyrosine kinase, protein kinase C, c-myc mRNA

expression and bcl-2 mRNA expression (Chen and Huang, 1998). Curcumin induces apoptotic

cell death by DNA-damage in human cancer cell lines, TK-10, MCF-7 and UACC-62 by acting

as topoisomerase II poison (Martin et. al., 2003). Recently, curcumin has been shown to cause

apoptosis in mouse neuro 2a cells by impairing the ubiquitin-proteasome system through the

mitochondrial pathway. Curcumin causes rapid decrease in mitochondrial membrane potential

and release of cytochrome c to activate caspase 9 and caspase 3 for apoptotic cell death (Jana et.

al, 2004). Recently, an interesting observation was made regarding curcumin-induced apoptosis

19

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in human colon cancer cell and role of heat shock proteins (hsp) thereon. In this study, SW480

cells were transfected with hsp 70 cDNA in either the sense or antisense orientation and stable

clones were selected and tested for their sensitivity to curcumin. Curcumin was found to be

ineffective to cause apoptosis in cells having hsp 70, while cells harbouring antisense hsp 70

were highly sensitive to apoptosis by curcumin as measured by nuclear condensation,

mitochondrial transmembrane potential, release of cytochrome c, activation of caspase 3 and

caspase 9 and other parameters for apoptosis (Rashmi et. al, 2004). Expression of glutathione S-

transferase Pl-1 (GSTPl-1) is correlated to carcinogenesis and curcumin has been shown to

induce apoptosis in K562 leukaemia cells by inhibiting the expression of GSTPl-1 at

transcription level (Duvoix et. a/.,2003). The mechanism of curcumin induced apoptosis has also

been studied in Caki cells, where curcumin causes apoptosis through downregulation of Bcl-XL

and LAP, release of cytochrome c and inhibition of Akt, which are markedly blocked by

A^acetylcysteine, indicating a role of ROS in curcumininduced cell death (Woo et. al, 2003). In

LNCaP prostrate cancer cells, curcumin induces apoptosis by enhancing tumour necrosis factor-

related apoptosis-inducing ligand (TRAIL). The combined treatment of the cell with curcumin

and TRAIL induces DNA fragmentation, cleavage of procaspase 3, 8 and 9, truncation of Bid

and release of cytochrome c from mitochondria, indicating involvement of both external

receptor- mediated and internal chemical-induced apoptosis in these cells (Deeb et. a/.,2003). In

colorectal carcinoma cell line, curcumin delays apoptosis along with the arrest of cell cycle at Gl

phase (Chen et. a/., 1996). Curcumin also reduces P53 gene expression, which is accompanied

with the induction of HSP-70 gene through initial depletion of intracellular Ca^ .̂ Curcumin also

produced nonselective inhibition of proliferation in several leukaemia, nontransformed

haematopoietic progenitor cells and fibroblast cell lines (Gautam et. a/., 1998). Curcumin induced

20

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apoptosis and large-scale DNA fragmentation have also been observed in Vg9Vdl+ T cells

through inhibition of isopentenyl pyrophosphate-induced NFA:B activation, proliferation and

chemokine production (Cipriani et. al, 2001). Curcumin inducesapoptosis in human leukaemia

HL-60 cells, which is blocked by some antioxidants (Kuo et. a/., 1996). Colon carcinoma is also

prevented by curcumin through arrest of cell-cycle progression independent of inhibition of

prostaglandin synthesis (Hanif et. a/., 1997). Curcumin suppresses human breast carcinoma

through multiple pathways. Its antiproliferative effect is estrogen dependent in ER (estrogen

receptor)-positive MCF-7 cells and estrogen-independent in ER-negative MDA-MB-231 cells.

Curcumin also downregulates matrix metalloproteinase (MMP)-2 and upregulates tissue

inhibitor of metalloproteinase (TIMP)-l, two common effector molecules involved in cell

invasion (Shao et. al, 2002). It also induces apoptosis through P53-dependent Bax induction in

human breast cancer cells38. However, curcumin affects different cell lines differently. Whereas

leukaemia, breast, colon, hepatocellular and ovarian carcinoma cells undergo apoptosis in the

presence of curcumin, lung, prostate, kidney, cervix and CNS malignancies and melanoma cells

show resistance to cytotoxic effect of curcumin (Khar et. a/.,2001). Curcumin also suppresses

tumour growth through various pathways. Nitric oxide (NO) and its derivatives play a major role

in tumour promotion. Curcumin inhibits iNOS and COX-2 production 69 by suppression of

NF)tB activation34. Curcumin also increases NO production in NK cells after prolonged

treatment, culminating in a stronger tumouricidal effect. Curcumin also induces apoptosis in AK-

5 tumour cells through upregulation (Khar et. al, 1999) of caspase-3. Reports also exist

indicating that curcumin blocks dexamethasone induced apoptosis of rat thymocytes (Sikora et.

al.,1997; Jaruga et. a/., 1998). Recently, in Jurkat cells, curcumin has been shown to prevent

glutathione depletion, thus protecting cells from caspase-3 activation and oligonucleosomal

f

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Review of Citerature

DNA fragmentation (Piwocka et. al., 2001). Curcumin also inhibits proliferation of rat

thymocytes (Sikora et. al, 1997).

3.2. Review on Zingiber officinale

Zingiber officinale Rose, is a slender, perennial rhizomatous herb. Leaves are linear, sessile and

glabrous. Flowers are yellowish green, arranged in oblong, cylindric spikes and ensheathed in a

few scarious, glabrous bracts. The rhizomes are white to yellowish brown in colour, irregularly

branched, somewhat armulated and laterally flattened. The growing tips are covered by a few

scales. The surface of the rhizome is smooth and if broken a few fibrous elements of the vascular

bundles project out from the cut ends (Warrier et. al, 1995). The plant is widely cultivated all

over India, Bangladesh, Taiwan, Jamaica and Nigeria. This perennial grows in warm climates.

Among natural food sources with antimicrobial activities, ginger ihizome (Z officinale Roscoe;

family Zingiberaceae) has been used as widely grown food spices and medicinal crops for

centuries. In particular, its pungent oil components harbour a series of polyphenolic ketones

called gingerols with many pharmacological activities. A few gingerol-related components have

been found to possess antibacterial and antifungal properties (Hiserodt et. al, 1998; Mahady et

al, 2003; Picker et. al, 2003). A broad range of biological activities have been attributed to Z

officinale. These include antiemetic (Mowrey and Clayson, 1982; Wood et. al., 1988; Bone et.

al, 1990), insecticidal (Roth et. al, 1998; Sahayaraj, 1998; Agarwal et. al, 2001), antibacterial

(Oloke et. al, 1989; Hiserodt et al, 1998), anUoxidant (Habsah et. al, 2000; Nakatini, 2000;

Sekiwa et. al, 2000), antirhinoviral (Denyer et. al, 1994), anti-inflammatory (Mascolo et. al,

1989) and antihepatotoxic (Hikino et. al, 1985) activities. Agarwal et. al, (2001) isolated five

compounds from Z officinale including gingerol, gingerone , dihydrogingerone,

22

"Review of Citerature

dehydrozingerone and dehydroshogaol, all of which were reported to significantly inhibit hyphal

growth of the potato pathogen Rhizoctonia solani. Although these studies identified compounds

with antifungal activities from ginger, human pathogens were not tested.

The ethanol and n-hexane extracts of ginger exhibited antibacterial activities against three

anaerobic Gram-negative bacteria, Porphyromonas gingivalis AT. CORDIFOLIAC 53978,

Porphyromonas endodontalis AT. CORDIFOLIAC 35406 and Prevotella intermedia AT.

CORDIFOLIAC 25611, causing periodontal diseases (Miri et. ai, 2008).

That the crude ethyl acetate extract, containing many compounds had excellent activity against a

diverse group of fungal pathogenic to humans. This was also reported by Agarwal et al. (2001)

where ginger oleoresin, a residue of acetone and ethanol extracts containing only 4 to 7.5%

gingerols and gingerol derivatives including shogaol, zingerone, paradols and gingerdiols was

reported to have antifungal activity against a plant pathogen similar to that of pure [6]-gingerol.

That results further indicate that Z. officinale can be used as an antifungal extractive enriched in

these compounds.

Z. officinale has been used to treat headaches, rheumatism, burns, peptic ulcer, dyspepsia,

depression and impotence (Tyler and Robbers, 1999). The oleoresin from the rhizome contain 6-

gingerol and its homolog's which have been shown to possess anti-inflammatory, antipyretic,

antihepatotoxic, analgesic and cardiotonic properties (Surh et. al., 1999). According to Mustafa

et. al., (1993), Z. officinale was found to inhibit the activity of cyclooxygenase and lipoxygenase

and hence decrease the pain in rheumatism and headaches. Topic et. al., (2002) also found that Z

officinale could inhibit lipid peroxidation by maintaining the levels of antioxidants in the serum

of rats treated with malathion. This is in concordance with an earlier study which found that 6-

gingerol in Z. officinale was a potent scavenger for the peroxyl radical which is the main product

23

"Review of Citerature

of lipid peroxidation (Aeschbach et. a/, 1994). Yamahara et. al, (1988) found that 6-gingerol

decreased the size of gastric lesions by 54.5%. Other studies had also found the antiemetic,

anticholesterolaemic and antiplatelet effects of Z officinale (Mowrey et. al., 1982; Tanabe et. al,

1993; Guh et. al, 1995). Mahady et. al, (2003) showed that, Ginger {Zingiber officinale Roscoe)

and the gingerols inhibit the growth of Cag A+ strains of Helicobacter pylori.

6-Shogaol of ginger, reduced the inflammatory response and protected the femoral cartilage from

damage produced in a CFA monoarthritic model of the knee joint of rats (Arkene et. al, 2006).

Ginger extracts rich in gingerols, shogaols, and other substances (gingerdiols, diaryltheptanoids,

zingiberene, arcurcumene, beta-bisabolene, neral, geranial, D-camphor, beta-phellandrene,

geranial, linalool, E-alpha-farnesene, beta-eudesmol) have recently gained considerable interest

for their capacity to interfere with cancer at initiation, promotion, and treatment (Aggarwal and

Shishodia, 2004). The gingerols (such as 6-, 8-, and 10-gingerols), a series of phenolic

compounds present in ginger root (ginger contains 1.0-3.0% gingerols), have been shown to

have chemopreventative effects that are associated with their antioxidative and antiinflammatory

activities (Katiyar et. al, 1996; Nagasawa et. al, 2002; Kim et. al, 2005; Surh et. al, 1999). The

rhizome of ginger (Zingiber offficinale Roscoe) has been used widely not only as a spice but also

as a useful crude drug in traditional Chinese medicine, including stomachic, antiemetic,

antidiarrheal and cardiotonic (Leung et. al, 1996). The constituents responsible for the pungent

taste of ginger are a homologous series of phenolic ketones, known as (Koo et. al., 2001)-, [6]-,

[8J-, [10]-, [12]-gingerols [2]. Among the total constituents, [6]-gingerol, an important pungent

component of ginger, was found to possess various pharmacological effects, such as cardiotonic,

antiemetic, antioxidant, antitumor, antiplatelet and anti-inflammatory [Suekawa et. a/., 1984, Koo

24

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et. a/.,2001, Ippoushi et. a/.,2003 and Tripathi et. a/.,2008]. In another finding, John and

Ojewole, (2006) reported that ethanol extract of Z. officinale rhizomes possesses analgesic, anti­

inflammatory and hypoglycaemic properties. This lends pharmacological support to folkloric,

ethnomedicinal uses of ginger in the treatment and/or management of painful, arthritic

inflammatory conditions, as well as in the management and or control of type 2 diabetes mellitus

in some rural Africa communities. It is used extensively in traditional Chinese medicine to treat

headaches, nausea, febrile conditions and colds; and in Ayurvedic and Western herbal medical

practices for the treatment of arthritis, rheumatic disorders and muscular discomfort (Dedov et.

ai, 2002; Jiang et. al, 2006). In South Africa, the fresh (or dried) rhizomes of Zingiber officinale

are used medicinally as stomachics and tonics to treat indigestion or dyspepsia, flatulence and

nausea (Van et. ai, 2002). The dried rhizomes of the herb (or extracts thereof) are also used

traditionally in other parts of the world for a variety of human ailments, including the treatment,

management and/or control of diarrhoea, dysentery, fever, cough, ulcers, boils and wounds.

Other reported pharmacological effects of Zingiber officinale rhizomes include antimicrobial,

analgesic, antipyretic, antiemetic, antiulcer, anxiolytic, cardiotonic, antihypertensive,

hypoglycaemic, antihyperlipidaemic, antiinflammatory and immuno-stimulant properties

(Suekawa et. ai, 1984; Mascolo et. al, 1989; Van Wyk et. al., 2004; Young et. al, 2005;

Ghayur and Gilani, 2005; Goyal and Kadnur, 2006; Jiang et al, 2006). 6-shogaol, the active

constituents of ginger downregulates inflammatory iNOS and COX-2 gene expression in

macrophages by inhibiting the activation of NFjB by interfering with the activation PI3K/Akt/IjB

kinases IKK and MAPK (Pan et. al, 2008).

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'Review of Citerature

Early animal studies had demonstrated the anti-emetic property of fresh ginger, but it was the

clinical work of Mowrey and Clayson which generated a wider interest in his use of ginger

(Mowrey and Clayson, 1982). They compared the effects of 1.88g of dried powdered ginger,

lOOmg dimenhydrinate (Dramamine) and placebo on the symptoms of motion sickness in 36

healthy subjects who reported very high susceptibility to motion sickness. Motion sickness was

induced by placing the blind folded subject in a tilted rotating chair. Ginger was found to be

superior to dimenhydrinate and placebo in preventing the gastrointestinal symptoms of motion

sickness and the authors postulated a local effort in the gastrointestinal tract for ginger. This was

particularly likely since it was given as a powder only 25 minutes before the test. The gingerols

and shogaols were subsequently identified as the main anti-emetic compounds in ginger (Kawai,

1994).

Early Chinese and Japanese research found that oral and intragastric application of fresh ginger

decoction produced a stimulant action on gastric secretion. German scientists found that chewing

9g of crystallised ginger had a profound effect on saliva production (Blumberger et. al., 1965).

Amylase activity was also increased and the saliva was less watery, although it contained slightly

less mucroprotein. Intraduodenal doses of ginger extract increased bile secretion in rats

(Yamaharaef. a/., 1985).

Gingerol and 10-gingerol were identified as the active components in given (Yamahara et. al.,

1985). Fresh ginger also contains a proteolytic enzyme (Thompson et. al., 1973). Ginger, in

conjunction with other pungent Ayurvedic herbs, increased the bioavailability of a number of

drugs by promoting their absorption and/or protecting them from being metabolized in their first

passage through the liver 1. Oral doses of 6-shogaol accelerated intestinal transit in rats (Suekawa

26

Jieview of Citerature

et. ai, 1984). Also an extract of ginger, and isolated 6-shogaol and gingerols, enhanced

gastrointestinal motility in mice after oral doses (Desai et. al., 1990).

Ginger and 6-gingerol inhibited experimental gastric ulcers in rats (Yamahara et. al., 1988; al-

Yahya, et. al., 1989). Fresh ginger decocted in water resulted in symptomatic improvement in 10

patients with peptic ulcers. Srivastava and co-workers found that aqueous extract of ginger

inhibited platelet aggregation induced by ADP, epinephrine, collagen and arachidonic acid in

vitro (Srivastava, 1984a). Ginger acted by inhibiting thromboxane synthesis (Srivastava, 1984b).

It also inhibited prostacyclin synthesis in rat aortal 1. The antiplatelet action of 6-gingerol was

also mainly due to the inhibition of thromboxane formation (Guh et. al., 1995).

Essential oil of ginger inhibited chronic adjuvant arthritis in rats (Mascolo et. al., 1989). Ginger

and its pungent components are dual inhibitors of arachiodonic acid metabolism. That is, they

inhibit both cyclooxygenase (prostaglandin synthetase) and lipoxygenase enzymes of the

prostaglandin and leukotriene biosynthetic pathways. Ginger extract given orally reduced fever

in rats by 38%, while the same dose of aspirin was effective by 44% (Mascolo et. ai, 1989). The

antipyretic activity of 6-shogaol and 6-gingerol has also been observed (Flynn et. ai, 1968).

Ginger exerted a powerful positive inotropic effect on isolated guinea pigs left atria (Shoji et. ai,

1982). Extracts of ginger have pronounced antioxidant activity comparable to that of synthetic

antioxidant preservatives (Govindarajan et. al., 1982).

Antihepatotoxic activities of gingerols and shogaols were observed using carbon tetrachloride

and galactosamine induced cytotoxicity in cultured rat hepatocytes (Hikino et. ai, 1985).

Injection of 6-shogaol showed an intense antitussive action in comparison with dihydrocodeine

phosphate (Naora et. ai, 1992).

27

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Two highly alkylated gingerols, [10]-gingerol and [12]-gingerol effectively inhibited the growth

of Porphyromonas gingivalis, Porphyromonas endodontalis and Prevotella intermedia, causing

periodontal diseases at a minimum inhibitory concentration (MIC) range of 6-30 microg/mL.

These ginger compounds also killed the oral pathogens at a minimum bactericidal concentration

(MBC) range of 4-20 microg/mL, but not the other ginger compounds 5-acetoxy-[6]-gingerol,

3,5-diacetoxy-[6]-Gingerdiol and galanolactone (Park et. al, 2008). [6]-gingerol inhibits

angiogenesis and may be useful in the treatment of tumors and other angiogenesis-dependent

diseases (Kim et.al, 2005).

One study reported that consumption of ginger through diet can confer protective effect against

the toxic effect of xenobiotics ( Nirmala et. al, 2010). The combinations of propolis extract +

clarithromycin and Z. officinale extract + clarithromycin have the potential to help control H.

pylori-associated gastroduodenal disease. (Nostro et. al., 2006).

Concomitant dietary feeding of ginger (l%w/w) significantly attenuated lindane-induced lipid

peroxidation, accompanied by modulation of Oxygen Free Radical scavenging enzymes as well

as reduced glutathione (GSH) and the GSH dependent enzymes glutathione peroxidase (Gpx),

glutathione reductase (GR) and glutathione-S-transferase (GST) in rats. (Rafat et. al., 2008).

Ginger supplementation suppressed liver carcinogenesis by scavenging the free radical

formation, and by reducing lipid peroxidation. (Yasmin et. al., 2009). 6-gingerol has two types of

antitumor effects: 1) direct colon cancer cell growth suppression, and 2) inhibition of the blood

supply of the tumor via angiogenesis. (Amy et. al., 2009). Z. officinale displayed some degree of

anti-schistosomal activity through reducing of the S. mansoni eggs output and the liver granuloma size.

(IsmaiUf. a/.,2007).

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"Review of Citerature

33. Review on Tinospora cordifolia

Tinospora cordifolia (Wild.) Miers ex Hook. F. & Thomas (T. cordifolia) being a rasayana drug

from Ayurveda is recommended for a number of diseases and for the promotion of health. It is

distributed throughout India as well as in China, Burma and Srilanka. There is evidence that the

plant is found in tropics of Africa and Australia (Singh et. ai, 1984). Its habitat ranges across a

wide region in India spreading from Kuraaon Mountains to Kanyakumari, the southern tip of

India. T. cordifolia has several vernacular names in various Indian languages: Gulvel (Marathi),

Giloe (Hindi), Gulancha (Bengali), Somida (Telugu), Sindal (Tamil) and Sittamrytu (Malyalam).

This plant has continued to draw the attention of research workers all over the world for over 50

years. The research work done on T. cordifolia ranges from experimental to clinical studies and

from phytochemistry to bioefficacy in diverse areas. A decision to review the available literature

was taken to determine if research carried out using modem methodologies has confirmed the

properties and uses described in traditional literature. It was thought that such an article would

serve as a useful database for future research and also enable experts in the field to determine if

the research on T. cordifolia is on right path.

Singh et. ai, (1984) have reported that an aqueous suspension of the alcohol extract of T.

cordifolia stem (300 mg/kg) exerted protection against CCU-induced liver damage (1.5 mL in

liquid paraffin v/v) in rats and mice. It maintained the hexobarbitone-induced sleeping time,

serum AST, ALT and ALP values as well as the hepatic architecture. It also significantly

reduced bromosulphathalein clearance time in CCU-treated rabbits. Post-treatment with T.

cordifolia (100 mg/kg, aqueous extract) for 15 days after development of liver injury (with 0.7

mL/kg of e c u ip- for 7 days) was found to be protective as demonstrated by a significant fall in

29

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AST, ALT, ALP and serum bilirubin (Bishayi et. al., 2002). In the above study, T. cordifolia was

used as a 'Satwa', which is a starchy polysaccharide preparation as recommended by Ayurveda

(Sharma, 2000). In addition, it reversed the immunosuppressive effects of CCI4 on rat peritoneal

macrophages.

Unlike the findings of previous workers, other studies have shown that treatment with T.

cordifolia (100 mg/kg) for 4 weeks prior to administration of a hepatotoxin such as CCI4 (1.25

mL/kg with liquid paraffin in 1:4 v/v) or galactosamine, resulted in aggravation of liver damage.

However, if used concurrently or following hepatotoxicity, it prevented fibrosis and preserved

the parenchymal cells (Rege et. al., 1984). In three different models of chronic liver damage (e.g.

liver damage due to free radicals, immune complex deposition and obstruction of the biliary

tract), fibrosis was found to be associated with altered activity of the reticuloendothelial system.

T. cordifolia was found to decrease fibrous tissue deposition and improve the functional activity

of the reticuloendothelial system (Nagarkatti et. al., 1994; Rege et. al., 1999). Additional

experimental studies showed that T. cordifolia reduced the incidence of endotoxemia in

cholestatic rats as evident from a bioassay. There was a reduction in mortality in the T. cordifolia

treated cholestatic rats given lead acetate (30% vs 100% in disease control; p < 0.05). It also

offered protection against complications of endotoxemia such as bacteremia, coagulopathy and

ischemic renal injury (Rege et. al., 1998). Co-administration of the aqueous extract of T.

cordifolia with antitubercular drugs was also found to decrease the histopathological scores

compared with the control group of animals with antitubercular drugs (Ambarkhane, 2004). In

studies with isolated primary hepatocytes and the HepG2 cell line, T. cordifolia increased the

proliferation of damaged hepatocytes exposed to 2.5 mM CCI4 compared with the disfilled water

control. T. cordifolia was also found to be cytotoxic to WEHl fibroblasts al a concentration of

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"Review of Citerature

1000 |ig/mL. However, it did not prevent the proliferation of fibroblasts when incubated directly

with these cells in nontoxic concentrations. The supernatants procured from macrophages after

incubating them with these concentrations of T. cordifolia extract were used to study their effects

on fibroblast proliferation. It was found that the supernatant obtained after incubating 800 |ig/mL

of T. cordifolia with macrophages prevented the proliferation of fibroblasts. Thus, the

antiproliferative activity of T. cordifolia was found to be secondary to the immune response

induced by T. cordifolia (Swar et.al, 2002). Studies using a standardized aqueous extract have

been reported in patients with hepatic disorders showing evidence of fibrosis and/or

immunosuppression. Maximum clinical exploration was done in patients with obstructive

jaundice, wherein an add-on regime of T. cordifolia (16 mg/kg/day) to conventional therapy

prior to surgical correction was found to reduce the mortality significantly [from 61.54% to 25%

in patients with PTBD (percutaneous transhepatic biliary drainage) and from 39% to 6.25% in

patients without PTBD. This was associated with a decrease in patients developing septicemia in

the T. cordifolia treated group. The depressed phagocytic activity of the polymorphonuclear cells

in patients with obstructive jaundice was found to improve after T. cordifolia therapy (Rege et.

ai, 1993). Serum GM-CSF levels were also increased as detected by the soft agar assay

technique. As evident from these studies, patients treated with T. cordifolia exhibited a better

quality of life in terms of reduced fullness of the abdomen, increased appetite, decreased nausea

and a sense of wellbeing. T. cordifolia was found to normalize the prolonged GI transit time

observed in obstructive jaundice. This may be secondary to the choleretic effect observed in

earlier studies (Singh et. al., 1984; Rege et. al., 1998). Pilot studies were conducted in a small

number of asymptomatic carriers of hepatitis B antigen (HBsAg) and cirrhotic patients to judge

the potential benefits of T. cordifolia. T. cordifolia therapy for 2 months in asymptomatic carriers

31

Jieview of Citerature

of hepatitis B antigen showed a three times higher rate of seroconversion (37.5%) than in the

placebo group (11.1%). These patients also showed an improvement in the intracellular killing

capacity of monocytes (51.67 ± 6.5% vs 38.38 ± 12.06% in placebo). In patients with cirrhosis,

therapy with T. cordifolia improved the Child-Pugh scores, shortened the prolonged antipyrine

half-life and increased the phagocytic and killing capacity of monocytes (Rege et. al., 1999a).

However, well-designed trials in larger study populations are needed to confirm these positive

trends. Two formulations containing T. cordifolia {Hemoliv and HP-1) have been documented to

have protective effects against CCU-induced hepatic damage in rats (Bhattacharya et. al, 2003;

Tasduq et. al, 2003). Another formulation, (Enliv), has demonstrated a protective effect in

broiler chickens given paracetamol as the damaging agent (Bhar et. al., 2005).

The effectiveness of T. cordifolia in models of infections and the lack of any evidence for

antimicrobial activity coupled with its properties and uses mentioned in the Ayurvedic textbooks

led to the hypothesis that the multiple actions ascribed to T. cordifolia may be mediated through

stimulation of immune cells and the subsequent cascade of events. Keeping in mind the role of

the psycho-neuroendocrino-immune axis, the effects of T. cordifolia on the immune system in

normal conditions and in several in vitro and in vivo models of immune suppression were

explored in depth (Dahanukar et. al., 1999). Based on the results, a standardized aqueous extract

of T. cordifolia was made available in the Indian market as an indigenous immunostimulant

(Dahanukar et. al., 1999; Rege et. al., 1999). In both normal and neutropenic mice, the whole

aqueous extract of T. cordifolia has been shown to cause leukocytosis with neutrophilia in a

dose-dependent manner. These findings have been confirmed by others (Manjrekar et. al., 2000).

T. cordifolia also stimulated the phagocytosis and killing activity of polymorphonuclear cells in

vitro as well as in vivo. The maximum effect on oral administration was seen on day 15 at 100

32

^Review of Citerature

mg/kg and on day 7 at 200 mg/kg. These doses have been shown to increase the proliferative

fraction of bone marrow cells, which explains the leukocytosis. Higher doses of T. cordifolia

(400 and 800 mg/kg) did not show a further increase in the WBC count and polymorphonuclear

cell functions and if therapy is continued beyond 7 days, the effects were not maintained. These

doses also increased apoptosis in normal bone marrow cells (Dahanukar et. al, 1999). In an in

vitro agarose gel assay, 200 ng/mL of T. cordifolia also produced a significant increase in the

chemotactic activity of PMN towards zymosan. The higher concentrations, however, did not

further increase the chemotactic activity (Rege et. al, 2002). An aqueous extract of T. cordifolia

has also been shown to stimulate phagocytosis and the candidicidal activity of peritoneal and

alveolar macrophages. T. cordifolia also exerted the same effect on human monocytes in vitro. It

augmented the clearance of particulate carbon by the reticuloendothelial system of rats. T.

cordifolia (100 mg/kg/day) has been shown to induce significantly colony forming units when

compared with distilled water (Thatte et. al, 1994). GM-CSF secreted by activated macrophages

also explains the stimulant action on polymorphonuclear functions. It has also been shown that if

activation of macrophages by T. cordifolia is blocked, the rise in WBC count is prevented. This

explains that macrophage activation by T. cordifolia plays a key role in its non-specific immune

effects (Rege et. al, 1996). An aqueous extract of T. cordifolia in concentrations ranging from

100 \i%lmL to 1000 ^g/mL, significantly increased nitric oxide (NO) release from unstimulated

and lipopolysaccharide (LPS) stimulated alveolar macrophages in an in vitro study. The effect

was maximum at a concentration of 200 ng/mL. Alveolar macrophages from the rats given T.

cordifolia extract (100 and 200 mg/kg/day for 15 days) also showed an increase in NO release,

both in unstimulated and LPS stimulated form, compared with the distilled water control. No

significant difference was observed between the two doses. Similar effects were seen when a

33

'Review of Citerature

murine macrophage cell line (RAW264.7) was used. A formulation of T. cordifolia containing a

standardized aqueous extract also evoked a significant rise in serum NO levels on day 21

compared with placebo (Vaingankar et.al, 2001). Recently, Singh et. al, (2006) have

demonstrated that an alcohol extract of the whole plant of T. cordifolia activated the antigen

presenting ability and phagocytosis of macrophages. Administration of T. cordifolia to young

mice also increased the thymocyte counts significantly. In T. cordifolia pre-treated mice, the

lymphopenic effect of hydrocortisone (125 mg/ kg i.p.) was blunted (Dahanukar et. al, 1999).

However, it did not exert any significant effect on the skin allograft reaction (Atal et. al, 1986).

The cytokine releasing properties of T. cordifolia have been recently studied by Dhuley et. al,

(1997) who used an alcohol extract of T. cordifolia (100 mg/kg) along with Ochratoxin A (OTA)

(1.5 mL/kg) in rats for a period of 17 weeks. Further studies have shown that in vivo

administration of an alcohol extract of the whole plant of T. cordifolia induces reactive nitrogen

intermediates, TNF-a and IL-1. It also exhibits a significant cytotoxicity (Singh et. al, 2004).

Sainis et. al, (1997) have conducted various experiments, which have led to the identification of

another target immune cell population for T. cordifolia, the B lymphocytes. They have shown

that an aqueous extract of the T. cordifolia stem exerts a concentration dependent polyclonal

mitogenic activity against murine spleen cells and nodes. Mature B lymphocytes appeared to be

more responsive than the precursors in the bone marrow. Using this bioassay, based on

mitogenic activity, fractionation of an aqueous extract has been carried out to isolate polyclonal

B cell activators in T. cordifolia, which are probably polysaccharides in nature (Sainis et al,

1997). Two preparations, partially purified immunomodulator (PPI) and its purified constituent

Gl-4 A, have been identified. It was also found that the aqueous extract inhibits rL-4 and IL-10

in supernatants from spleen cells (Sainis et al, 1999). The purified polysaccharide isolated by

34

"Review of Citerature

this research group has been characterized as an acidic arabinogalactan polysaccharide

(Chintalwar et. ai, 1999). The novel a-D-glucan (RRl) isolated from the T. cordifolia stem has

been shown to activate different subsets of lymphocytes, especially NK cells and increased

synthesis of cytokines associated with However, it did not produce any oxidative stress or

increased nitric oxide production (Nair et. ai, 2004). Further work with this glucan has revealed

that it stimulates the immune system by activating the macrophages through TLR 6 signalling,

and NF-KB activation mechanism, leading to the production of cytokines and chemokines (Nair

et. ai, 2006). Kapil and Sharma, (1997) have characterized the immunopotentiating properties of

various isolated phytochemical constituents from T. cordifolia. Apart from the clinical studies

conducted in patients with hepatic disorders having evidence of immunosuppression, T.

cordifolia has been evaluated in other conditions wherein support for the immune system is

warranted. Ayurveda advocates the use of T. cordifolia for the treatment of tuberculosis

(Upadhyaya, 2000). The effect of T. cordifolia (1500 mg/day in divided doses for 6 months

given as an adjunct to chemotherapy) has been evaluated on the our. cordifoliaome of the

chemotherapeutic course of patients with pulmonary tuberculosis in two double-blind placebo

controlled randomized trials. The incidence of adverse events was less in this group and the

quality of life was better (Dash, 2000). T. cordifolia included in the standard management

protocol for burns significantly increased the WBC count and IgG levels by day 14 of therapy

compared with the placebo-treated control. The survival rate in both the groups was, however,

comparable (Dahanukar et ai, 1999). In a double blind placebo-control study, the effect of T.

cordifolia on the development of antibody to vaccine and GM-CSF levels was explored. T.

cordifolia was used in a dose of 1500 mg/ day (for a mar. cordifoliahed placebo) on day 1 of

hepatitis B immunization and thereafter was continued for 6 months (Vaingankar, 2001).

35

'Review of Citerature

Significantly higher anti-HBsAg litres were noted after 3 months of T. cordifolia therapy, which

were maintained until 6 months compared with the placebo group {p < 0.01). GM-CSF levels

were also significantly higher in the treated group (p < 0.05 vs placebo group). T. cordifolia may

have a role to play as a potential vaccine adjuvant in future.

Diverse modes of action of T.cordifolia like activation of macrophages (attenuation of radiation-

induced decrease in adherence and spreading), reduced apoptosis and increase cell proliferation

and increased IL-ip and GM-CSF levels may be responsible for impressive radioprotective

efficacy (Singh et. al, 2007).

The aqueous extract of stem of T. cordifolia has been shown to decrease fasting blood sugar

levels and increase glucose tolerance (Gupta et. al, 1967). It also caused a significant fall in

blood glucose levels in alloxan-induced hyperglycemia (Raghunathan and Sharma, 1969). When

an aqueous extract of T. cordifolia root was tested in doses of 2500 mg/kg, 5000 mg/kg and 7500

mg/kg administered orally in alloxan-induced (150 mg/kg i.p.) diabetic rats (Stanely et. al.,

2000), the 5000 mg/kg dose was most effective. Similar effects have been described with the

ethanol extract of T. cordifolia root (5000 mg/kg), which produced hypoglycemia and decreased

serum and tissue lipids (Prince and Menon, 2003). Grover et. al.,. (2000) have used the same

model but with increasing doses of alloxan (32 mg/kg i.v., 120 mg/kg i.v. and 150 mg/kg i.p.) to

induce mild, moderate and severe hyperglycemia. The effect of T. cordifolia (400 mg/kg of

aqueous extract) was maximum against mild hyperglycemia, less against moderate

hyperglycemia and poor against severe hyperglycemia. It was hence postulated that the

antihyperglycemic effect of T. cordifolia might be predominantly due to an increase of insulin

release from the beta cells and not due to a direct insulinomimetic action. In another study

Sengupta et. al., (2009) suggested that T. cordifolia have protective effect on lipid peroxidation

36

Tieview of Citerature

and antioxidative enzymes in streptozotocin-induced diabetic rats. An ethanol extract of the T.

cordifolia stem (250 mg/kg) also lowered blood glucose level within 1 week in alloxan-induced

(100 mg/kg i.p.) diabetic rats but the effect was less than that of other plants tested using the

same model (Kar et al., 2003). T. cordifolia has been shown to increase glycolysis and to

increase the glucose 6-phosphate concentration in diabetic rats (Grover et. al., 2000; Stanely et.

al, 2000). A significant antihyperlipidemic action of T. cordifolia root extract at a dose of 5000

mg/kg in alloxan-induced diabetic rats has been reported (Prince et. al, 1999). The aqueous

extract of T. cordifolia at a dose of 5000 mg/kg, the aqueous extract of T. cordifolia decreased

the ThioBarbituric Acid Reactive Substances (TBARS) in the liver and kidney while increasing

the TBARS in the heart. Also, the glutathione (GSH) and superoxide dismutase (SOD) values in

the liver and kidney were significantly lower compared with the diabetic controls (Prince and

Menon, 2001). In further studies, this team established that as observed with the aqueous extract,

the alcohol extract of T. cordifolia root (100 mg/kg) when given orally for 6 weeks following 2

weeks of alloxan-induced diabetes also normalized the antioxidant status of the liver and kidney

(Prince et. a/., 2004). Recently, an herbal formulation containing T. cordifolia (SH-OID) was

tested in an animal study along with agents causing insulin resistance, namely, dexamethasone

and^fructose (Shalam et. al, 2006). It not only prevented the rise in serum glucose, serum

cholesterol, LDL and triglycerides, but also improved the insulin sensitivity. The reductions in

body weight and liver glycogen levels were also prevented. Sarma et. al, (1996) investigated the

anti stress activity of an ethanol extract of T. cordifolia (100 mg/kg/day) in various animal

models simulating stress and related depression. The onset and duration of immobility after an

initial period of vigorous activity in the forced swimming endurance test have been suggested to

represent depression following stress. Pre-treatment with T. cordifolia for 10 days followed by

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'Revie'w of Citerature

administration of the above test, significantly delayed the onset and reduced the duration of

immobility. When analgesiometer measurements were taken following a similar treatment

schedule, T. cordifolia exerted a significant analgesic effect (Sarma et ai, 1996). T. cordifolia

appears to modulate the psychoneuroendocrine axis through its immunostimulant activity and to

offer protection against various stressors (Rege et. ai, 1999). Further studies showed that

prevention of cold immobilization stress induced gastric mucosal damage by an aqueous extract

of T. cordifolia was associated with a prevention in the rise of liver malondialdehyde and

depletion of liver GSH content in T. cordifolia treated rats (Sheth, 2002). In an in vitro study, 10

jxg/mL of methanol extract of T. cordifolia stem inhibited microvessel outgrowth from the rat

aorta ring induced by the conditioned medium from the B16F10 melanoma cells. The

conditioned medium obtained from extract treated B16F10 melanoma cells also prevented vessel

outgrowth from aortic rings (Leyon and Kuttan, 2004a). The polysaccharide fraction of T.

cordifolia isolated using the protocol of Chintalwar et. ai, (1999) when injected

intraperitoneally (0.5 mg/animal/day for 10 days) in syngeneic C57BL/6 mice along with tumor

challenge (B16F-10 melanoma cells; 106 cells/0.5 mL PBS i.v.), inhibited the number of lung

metastatic colonies giving 72% inhibition compared with the vehicle control and hence it was

postulated that T. cordifolia may prevent lung metastases by modulating B cells or non-specific

immune cells such as NK cells (Leyon and Kuttan, 2004b). A study reported by Ayurvedic

practitioners demonstrates the antipyretic effect of T. cordifolia individually and as concomitant

therapy with paracetamol and/or antibiotics (Kumar and Shrivastav, 1995). Ayurveda states that

T. cordifolia has antiinflammatory properties and is used in therapy of rheumatoid arthritis and

gout (Gogte, 2000). Pendse et. ai, (1977) have shown that T. cordifolia stem (aqueous extract,

50 mg/100 g, p.o. and i.p.) exhibits a significant reduction in carrageenin-induced acute

38

'Review of Citerature

inflammation. The percent reduction in edema both by oral and intraperitoneal administration

(49.12% and 63.16%) were significant (p < 0.05) but less when compared with the

dexamethasone given i.p. (95.9%). Similar results have been reported recently for a 50 mg/ kg

suspension of pulverized dried stem of T. cordifolia using the same model (67.28% decrease; p <

0.001) (Jana et. ai, 1999). In this latter study, the antiinflammatory effect of T. cordifolia was

reported to be more than acetylsalicylic acid (100 mg/kg). The aqueous extract of T. cordifolia

root has also been found to inhibit the rat paw edema induced by carrageenin (Pendse et. ai,

1981). The authors have postulated that the anti inflammatory effect of T. cordifolia was due to

interference with histamine and serotonin release. Pendse et. ai, (1977) have also reported that in

doses of 20 mg/100 g and 10 mg/100 g p. o., respectively, T. cordifolia significantly prevented

granulation tissue formation in the granuloma pouch technique (using croton oil) and

demonstrated a significant decrease in primary and secondary responses to Freunds adjuvant. T.

cordifolia was found to decrease histamine-induced bronchospasm in guinea-pigs. It reduced the

capillary permeability in mice and also reduced the mast cell disruption in rats (Nayampalli et.

ai, 1986).A randomized, double-blind, placebo-controlled trial has been conducted in patients

with allergic rhinitis to whom an aqueous extract of T. cordifolia stem was administered. T.

cordifolia not only reduced the symptoms of allergic rhinitis but also reduced the eosinophil and

goblet cell count in the nasal smears. This was associated with an increase in the total leukocyte

count (Badar et. ai, 2005). A study reported by Ayurvedic practitioners demonstrates the

antipyretic effect of T. cordifolia individually and as concomitant therapy with paracetamol

and/or antibiotics. Although T. cordifolia given alone exerted an antipyretic effect comparable to

that of the other groups in the study, the number of patients/group (8/ group) was too few to

39

lieview of Citerature

comment on its significance. Further, the article does not mention the antibiotics used (Kumar

and Shrivastav, 1995).

40