Antioxidant, Anti-inflammatory and Antinociceptive Activities of Essential Oil from Ginger.

Chemical compositions of ginger oil as well as its antioxidant, anti-inflammatory and antinociceptive potential were evaluated in the present study. The main constituents as detected by GC/MS analysis was V-zingiberene which constituted 31% of the total area, ar-curcumene (15.4%) and a -sesquiphellandrene (14.02%). Ginger oil scavenged superoxide, DPPH, hydroxyl radicals and inhibited tissue lipid peroxidation in vitro . Intraperitoneal administration of ginger oil was found to inhibit phorbol- 12-myristate-13-acetate induced superoxide radicals elicited by macrophages. Oral administration of ginger oil for one month, significantly increased superoxide dismutase, glutathione and glutathione reductase enzymes level (P<0.001) in blood of mice and glutathione-S-transferase, glutathione peroxidase and superoxide dismutase enzymes in liver. Ginger oil produced significant reduction in acute inflammation produced by carrageenan and dextran and formalin induced chronic inflammation (P<0.001). It also exhibited significant reduction in acetic acid induced writhing movements (P<0.001). The present report revealed that ginger oil possesses antioxidant activity as well as significant anti-inflammatory and antinociceptive property.

Hepato-protective potential of carotenoid meso-zeaxanthin against paracetamol, CCl4 and ethanol induced toxicity.

Hepato-protective potential of carotenoid meso-zeaxanthin [(3R, 3’S)-, -carotene-3, 3′-diol] was studied using in vivo rat models. Paracetamol (3 g/kg body wt, orally), 20% ethanol (7.5 g/kg body wt, orally) and CCl4 (2.5ml /kg, ip) were used as hepato toxins. Levels of marker enzymes of hepatic injury such as serum glutamate oxaloacetate transaminase, serum glutamate pyruvate transaminase and alkaline phosphatase, and serum bilirubin, which were drastically elevated by these hepato toxins were significantly decreased by meso-zeaxanthin pretreatment in a dose-dependent manner. Oxidative stress markers, tissue lipid peroxidation, conjugated dienes and tissue hydroperoxides, were high in the paracetamol treated control group animals, which were lowered by meso-zeaxanthin administration. Level of glutathione and antioxidant enzymes, superoxide dismutase, catalase and glutathione peroxidase, in liver tissue was increased by meso-zeaxanthin pretreatment compared to control group during alcohol and CCl4 induced hepatotoxicity. Hydroxyproline, an indicator of fibrosis in liver tissue, decreased remarkably by meso-zeaxanthin administration despite its notable elevation in ethanol treated rats. Histopathological analysis of liver tissue showed the hepatoprotective potential of meso-zeaxanthin.

Antioxidant activity of carotenoid lutein in vitro and in vivo.

Carotenoid lutein was evaluated for its antioxidant potential both in vitro and in vivo. Lutein was found to scavenge superoxide radicals, hydroxyl radicals and inhibited in vitro lipid peroxidation. Concentrations needed for 50 % inhibition (IC50) were 21, 1.75 and 2.2 g/mL respectively. It scavenged 2,2-diphenyl-1-picryl hydrazyl (IC50 35 g/mL) and nitric oxide radicals (IC50 3.8 g/mL) while 2,2-azobis-3-ethylbenzthiozoline-6-sulfonic acid radicals were inhibited at higher concentration. Ferric reducing power (50%) of lutein was found to be equal 0.3μmols/mL of FeSO4.7H2O. Its oral administration inhibited superoxide generation in macrophages in vivo by 34.18, 64.32 and 70.22 % at doses of 50, 100 and 250 mg/kg body weight. The oral administration of lutein in mice for 1 month significantly increased the activity of catalase, superoxide dismutase, glutathione reductase and glutathione in blood and liver while the activity of glutathione peroxidase and glutathione-S-transferase were found to be increased in the liver tissue. Implication of these results in terms of its role in reducing degenerative diseases is discussed.

Antidiabetic and antioxidant activity of Terminalia belerica. Roxb.

Effect of continuous administration of dried 75% methanolic extract of fruits of Terminalia belerica (Combretaceae) suspended in water was studied in alloxan induced hyperglycemia and antioxidant defense mechanism in rats. T. belerica prevented alloxan-induced hyperglycaemia significantly from 6th day of administration and there was 54% reduction on 12th day. Oxidative stress produced by alloxan was found to be significantly lowered by the administration of T. belerica extract. This was evident from a significant decrease in thiobarbituric acid reactive substances, conjugated dienes and hydroperoxides in blood and liver respectively. Similarly, decreased glutathione level produced by alloxan was increased by the administration of the extract in blood and liver. However the increase was not significant. Superoxide dismutase which was decreased by alloxan was significantly increased from 9th day in blood and liver of drug treated group. Similarly there was significant increase in the activity of catalase in blood and liver. Decrease in glutathione peroxidase by alloxan administration was found to be increased significantly in the blood and liver from 9th day by extract treatment. Glutathione reductase also was found to be increased in blood and liver. These results suggested that T. belerica fruit extract possessed anti-diabetic and anti-oxidant activity and these activities may be interrelated.

Hepato and reno protective action of Calendula officinalis L. flower extract.

Flower extract of C. officinalis L. was evaluated for its protective effect against CCl4 induced acute hepatotoxicity and cisplatin induced nephrotoxicity. The activities of serum marker enzymes of liver injury like glutamate pyruvate transaminase (SGPT), glutamate oxaloacetate transaminase (SGOT) and alkaline phosphatase (ALP) which were increased by CCl4 injection was found to be significantly reduced by the pretreatment of the flower extract at 100 and 250 mg/kg body weight. The lipid peroxidation in liver, the marker of membrane damage and the total bilirubin content in serum were also found to be at significantly low level in the extract pretreated group, indicating its protective role. The kidney function markers like urea and creatinine were significantly increased in cisplatin treated animals. However, their levels were found to be lowered in the extract pretreated groups (100 and 250 mg/kg body weight). Moreover, cisplatin induced myelosuppression was ameliorated by the extract pretreatment. Treatment with the extract produced enhancement of antioxidant enzymes--superoxide dismutase and catalase and glutathione. Results suggest a protective role of the flower extract of C. officinalis against CCl4 induced acute hepatotoxicity and cisplatin induced nephrotoxicity. Extract has been found to contain several carotenoids of which lutein, zeaxanthin and lycopene predominates. Possible mechanism of action of the flower extract may be due to its antioxidant activity and reduction of oxygen radicals.

Anti-inflammatory activity of flower extract of Calendula officinalis Linn. and its possible mechanism of action.

Calendula officinalis flower extract possessed significant anti-inflammatory activity against carrageenan and dextran-induced acute paw edema. Oral administration of 250 and 500 mg/kg body weight Calendula extract produced significant inhibition (50.6 and 65.9% respectively) in paw edema of animals induced by carrageenan and 41.9 and 42.4% respectively with inflammation produced by dextran. In chronic anti-inflammatory model using formalin, administration of 250 and 500 mg/kg body weight Calendula extract produced an inhibition of 32.9 and 62.3% respectively compared to controls. TNF-alpha production by macrophage culture treated with lipopolysaccharide (LPS) was found to be significantly inhibited by Calendula extract. Moreover, increased levels of proinflammatory cytokines IL- 1beta, IL-6, TNF-alpha and IFN-gamma and acute phase protein, C- reactive protein (CRP) in mice produced by LPS injection were inhibited significantly by the extract. LPS induced cyclooxygenase-2 (Cox-2) levels in mice spleen were also found to be inhibited by extract treatment. The results showed that potent anti-inflammatory response of C. officinalis extract may be mediated by the inhibition of proinflammatory cytokines and Cox-2 and subsequent prostaglandin synthesis.

Clastogenic potential of Ruta graveolens extract and a homeopathic preparation in mouse bone marrow cells.

Ruta graveolens belonging to family Rutaceae has long been traditionally used as a medicinal plant as well as a flavoring agent in food. However, very little data are available on the toxicity of the plant. This report presents evidence on the genotoxic and clastogenic potential of an extract of Ruta graveolens and Ruta 200C, a homeopathic preparation. Various types of chromosomal aberrations were noted in bone marrow cells after treatment. The percentage of aberrated cells in the 400mg/kgb.wt extract administered group was found to be 21% and with 1,000 mg/kg.b.wt it was 31%. The value for the Ruta 200C treated group was also elevated to 23% as compared to the 3%for untreated animals. In addition, bone marrow cells had higher incidence of micronuclei induction when treated with the extract (400 mg and 1,000 mg/kg body weight) and Ruta 200C for 30 days. Administration of the extract (1,000 mg/kg.b.wt) over a period of 30 days also resulted in damage to cellular DNA as evidenced by comet formation where the comet parameters such as percentage DNA in tail, tail length, tail moment of the bone marrow cells were increased several fold over control values. The comet tail moment of the bone marrow cells increased from 4.5 to 50.2 after the extract treatment. Administration of Ruta 200C for 5 consecutive days increased the tail moment to 11.7. These results indicate that Ruta graveolens and Ruta 200C may induce genotoxicity in animals.

Immunostimulatory action of AC II--an ayurvedic formulation useful in HIV.

Immunostimulatory activity of AC II, a registered ayurvedic preparation prepared at Amala Ayurvedic Research Centre for treating HIV and AIDS is reported. AC II administration could significantly enhance the mitogen-induced proliferation of lymphocytes of spleen cells. It was also found to increase cell-mediated immune responses in normal and tumor-bearing control animals. Oral administration of AC II significantly enhanced Natural Killer cell activity in normal and tumor-bearing animals on the 7th day, which was observed earlier than the tumor-bearing control animals and normal animals. Antibody dependent cellular cytotoxicity (ADCC) was also increased in AC II treated normal and tumor-bearing animals. An early enhancement of antibody-dependent complement-mediated cytotoxicity was also observed by the administration of AC II in normal as well as tumor-bearing animals. Treatment with AC II elevated the levels of IL-2, TNF-alpha and IFN-gamma in normal mice. Administration of AC II was also found to increase the cytotoxic T lymphocyte production in EL4 treated mice. These studies support the use of this immune stimulatory preparation in HIV patients.

Effect of homeopathic medicines on transplanted tumors in mice.

Ultra low doses used in homeopathic medicines are reported to have healing potential for various diseases but their action remains controversial. In this study we have investigated the antitumour and antimetastatic activity of selected homeopathic medicines against transplanted tumours in mice. It was found that Ruta graveolens 200c and Hydrastis canadensis 200c significantly increased the lifespan of Ehrlich Ascites Carcinoma and Dalton's Lymphoma Ascites induced tumour-bearing animals by 49.7%, and 69.4% respectively. Moreover there was 95.6% and 95.8% reduction of solid tumour volume in Ruta 200c and Hydrastis 200c treated animals on the 31st day after tumour inoculation. Hydrastis 1M given orally significantly inhibited the growth of developed solid tumours produced by DLA cells and increased the lifespan of tumour bearing animals. Some 9 out of 15 animals with developed tumors were completely tumour free after treatment with Hydrastis 1M. Significant anti-metastatic activity was also found in B16F-10 melanoma-bearing animals treated with Thuja1M, Hydrastis 1M and Lycopodium1M. This was evident from the inhibition of lung tumour nodule formation, morphological and histopathological analysis of lung and decreased levels of gamma-GT in serum, a cellular marker of proliferation. These findings support that homeopathic preparations of Ruta and Hydrastis have significant antitumour activity. The mechanism of action of these medicines is not known at present.

Inhibition of chemically induced carcinogenesis by drugs used in homeopathic medicine.

Homeopathy is considered as one modality for cancer therapy. However, there are only very few clinical reports on the activity of the drugs, as well as in experimental animals. Presently we have evaluated the inhibitory effects of potentized homeopathic preparations against N'-nitrosodiethylamine (NDEA) induced hepatocellular carcinoma in rats as well as 3-methylcholanthrene-induced sarcomas in mice. We have used Ruta, Hydrastis, Lycopodium and Thuja, which are commonly employed in homeopathy for treating cancer. Administration of NDEA in rats resulted in tumor induction in the liver and elevated marker enzymes such as gamma-glutamyl transpeptidase, glutamate pyruvate transaminase, glutamate oxaloacetate transaminase and alkaline phosphatase in the serum and in liver. Concomitant administration of homeopathic drugs retarded the tumor growth and significantly reduced the elevated marker enzymes level as revealed by morphological, biochemical and histopathological evaluation. Out of the four drugs studied, Ruta 200c showed maximum inhibition of liver tumor development. Ruta 200c and phosphorus 1M were found to reduce the incidence of 3-methylcholanthrene-induced sarcomas and also increase the life span of mice harboring the tumours. These studies demonstrate that homeopathic drugs, at ultra low doses, may be able to decrease tumor induction by carcinogen administration. At present we do not know the mechanisms of action of these drugs useful against carcinogenesis.

Effect of AC II, a herbal formulation on radiation-induced immunosuppression in mice.

A single dose of 6 Gy irradiation significantly reduced the total WBC count while in herbal formulation (AC II) treated groups it was found to be significantly increased. Similarly bone marrow cellularity and alpha-esterase positive cells, which were lowered by radiation, were partly restored in AC II treated groups. The data indicate that AC II can overcome the immunosuppression produced by irradiation.

Anti-tumour activity of Ruta graveolens extract.

An extract of Ruta graveolens was found to be cytotoxic to Dalton's lymphoma ascites (DLA), Ehrlich ascites carcinoma (EAC) and L929 cells in culture (IC100=16 mg/ml) and also to increase the lifespan of tumour bearing animals. The extract further decreased solid tumours developing from DLA and EAC cells when given simultaneously with elongation of the lifespan of tumour-bearing animals. A homeopathic preparation of Ruta graveolens (200 c) was equally effective. Neither was effective for reducing already developed tumours. The Ruta graveolens extract was found to scavenge hydroxyl radicals and inhibit lipid peroxidation at low concentrations. However, at higher concentrations the extract acted as a prooxidant as inhibition of lipid peroxidation and scavenging of hydroxyl radical was minimal. These data indicates that the prooxidant activity of Ruta graveolens may be responsible for the cytocidal action of the extract and its ability to produce tumour reduction.

Inhibition of N-Methyl N'-nitro-N-nitrosoguanidine (MNNG) induced gastric carcinogenesis by Phyllanthus amarus extract.

Chemopreventive activity of Phyllanthus amarus Schum & Thonn (Euphorbiaceae) extract was studied with regard to N-methyl N'-nitro-N-nitrosoguanidine (MNNG) induced stomach cancer in Wistar rats. Administration of the extract with MNNG significantly reduced the incidence of gastric neoplasms in rats (44%) as well as their numbers. Moreover, elevated levels of enzymes in the stomach were found to be reduced by P. amarus administration. For example, gamma-glutamyl transpeptidase activity was decreased from 20.3 +/- 6.7 mmol/min/mg protein to almost normal levels (2.8 +/- 0.9) by 750 mg/kg body weight of the extract. Similarly glutathione S-transferase activity (1317.6 +/- 211 n mol/min/mg protein) and glutathione reductase (368 +/- 66) levels in the MNNG treated group were found to be lowered to 494.8 +/- 76 and 192 +/- 45, respectively, while reduced glutathione (GSH) was increased from 4.6+/- 0.9 to 8.5+/-1.4 n mol/min/mg protein. AgNOR dots and clusters, indicators of cellular proliferation, which were increased by MNNG treatment, became near to normal in P. amarus treated animals.

Antidiabetic activity of Aegle marmelos and its relationship with its antioxidant properties.

Oxidative stress induced by alloxan has been shown to damage pancreatic beta-cell and produce hyperglycemia in rats. Aegle marmelos leaf extract is being used in Ayurveda as a medicine for diabetes. The present study examined the action of Aegle marmelos against experimental diabetes as well as the antioxidant potential of the drug. A methanolic extract of Aegle marmelos was found to reduce blood sugar in alloxan diabetic rats. Reduction in blood sugar could be seen from 6th day after continuous administration of the extract and on 12th day sugar levels were found to be reduced by 54%. Oxidative stress produced by alloxan was found to be significantly lowered by the administration of Aegle marmelos extract. This was evident from a significant decrease in lipid peroxidation, conjugated diene and hydroperoxide levels in serum as well as in liver induced by alloxan. Catalase and glutathione peroxidase activity in blood and liver were found to be increased from 9th day onwards after drug administration. Superoxide dismutase and glutathione levels were found to be increased only on 12th day. These results indicate that Aegle marmelos extract effectively reduced the oxidative stress induced by alloxan and produced a reduction in blood sugar.

Hypoglycemic effect of methanol extract of Phyllanthus amarus Schum & Thonn on alloxan induced diabetes mellitus in rats and its relation with antioxidant potential.

Methanolic extract of P. amarus was found to have potential anti-oxidant activity as it could inhibit lipid peroxidation, and scavenge hydroxyl and superoxide radicals in vitro. The amount required for 50% inhibition of lipid peroxide formation was 104 microg/ml and the concentrations needed to scavenge hydroxyl and superoxide radicals were 117 and 19 microg/ml respectively. The extract was found to reduce the blood sugar in alloxan diabetic rats at 4th hr by 6% at a dose level of 200 mg/kg body wt and 18.7% at a concentration of 1000 mg/kg body wt. Continued administration of the extract for 15 days produced significant (P < 0.001) reduction in blood sugar. On 18th day after alloxan administration values were almost similar to normal in the group taking 1000 mg/kg body wt.

Immunopotentiating activity of abrin, a lectin from Abrus precatorius Linn.

A non-toxic dose of abrin, (1.25 microg/kg body wt) consecutively for five days in normal mice stimulated specific humoral responses. A noticeable increase was observed in total leucocyte count, lymphocytosis, weights of spleen and thymus, circulating antibody titre, antibody forming cells, bone marrow cellularity and alpha-esterase positive bone marrow cells. The results suggest that abrin can potentiate the humoral immune response of the host.

Antitumour effect of abrin on transplanted tumours in mice.

Abrin, a galactose specific lectin was purified using sepharose 4B affinity column from seeds of Abrus precatorius. It exhibited antitumour activity in mice when used at a sublethal dose of 7.5 micrograms/kg every alternate day for 10 days. Both intralesional and intraperiloneal (i.p.) administration of abrin was effective in reducing solid tumour mass development induced by Dalton's Lymphoma Ascites (DLA) and Ehrlich's Ascites Carcinoma (EAC) cells. DLA cell line was more sensitive to abrin than EAC. Abrin when injected i.p. increased the life span of ascites tumour bearing mice. Abrin when used simultaneously with tumour cells brought about maximum antitumour effect. On developed tumour masses, abrin administration brought about significant reduction in tumour volume, especially in DLA induced tumours. Prophylactic administration of abrin was found ineffective.