Exploring the Potential Role of Chemopreventive Agent,Hesperetin Conjugated Pegylated Gold Nanoparticles in Diethylnitrosamine-Induced Hepatocellular Carcinoma in Male Wistar Albino Rats

Liver cancer is the fifth most common cancer and is still one of the leading causes of death world wide, due to food additives, alcohol, fungal toxins, air, toxic industrial chemicals, and water pollutants. Chemopreventive drugs play a potential role in liver cancer treatment. Obviously in the production of anticancer drugs, the factors like poor solubility, bioavailability, biocompatibility, limited chemical stability, large amount of dose etc., plays a major role. Against this backdrop, the idea of designing the chemopreventive nature of bio flavanoid hesperetin (HP) drug conjugated with pegylated gold nanoparticles to increasing the solubility, improve bioavailability and enhance the targeting capabilities of the drug during diethylnitrosamine (DEN) induced liver cancer in male wistar albino rats. The dose fixation studies and the toxicity of pure HP and HP conjugated gold nanoparticles (Au-mPEG₍₅₀₀₀₎-S-HP) were analysed. After concluded the dose fixation and toxicity studies the experimental design were segregated in six groups for the anticancer analysis of DEN induced HCC for 16 weeks. After the experimental period the body weight, relative liver weight, number of nodules and size of nodules, the levels of tumor markers like CEA, AFP and the level of lipid peroxidation, lipid hydroperoxides and the activities of antioxidant enzymes were assessed. The administration of DEN to rats resulted in increased relative liver weight and serum marker enzymes aspartate transaminase, alanine transaminase, alkaline phosphatase, lactate dehydrogenase, and gamma glutamyl transpeptidase. The levels of lipid peroxides elevated (in both serum and tissue) with subsequent decrease in the final body weight and tissue antioxidants like superoxide dismutase, catalase, reduced glutathione, glutathione peroxidise, and glutathione reductase. HP supplementation (20 mg/kg b.wt) significantly attenuated these alterations, thereby showing potent anticancer effect in liver cancer and the HP loaded gold nanoparticels (Au-mPEG₍₅₀₀₀₎-S-HP) treated animals shows the better treatment than the pure HP due to the solubility of drug, bioavailability and the target drug delivery of the biodegradable polymer. Histological observations were also carried out, which added supports to the chemopreventive action of the pure HP and HP loaded gold nanoparticles (Au-mPEG₍₅₀₀₀₎-S-HP) against DEN induction during liver cancer progression. These findings suggest that HP loaded gold nanoparticels (Au-mPEG₍₅₀₀₀₎-S-HP) shows better efficacy than the pure HP against lipid peroxidation, hepatic cell damage and protects the antioxidant system in DEN induced hepatocellular carcinogenesis.     

Potential Effect of <Emphasis Type="Italic">Bacopa monnieri</Emphasis> on Nitrobenzene Induced Liver Damage in Rats

The study was designed to evaluate the hepatoprotective activity of ethanolic extract of Bacopa monnieri in acute experimental liver injury induced by Nitrobenzene in rats. The extract at the dose of 200 mg/kg body weight was administered orally once every day for 10 days. The increased serum marker enzymes, Aspartate transaminase, Alanine transaminase and alkaline phosphatase were restored towards normalization significantly by the extract. Significant increase in SOD, CAT and GPx was observed in extract treated liver injured experimental rats. Histopathological examination of the liver tissues supported the hepatoprotection. It is concluded that the ethanolic extract of Bacopa monieri plant possess good hepatoprotective activity.     

Effect of thiazide drug compound on the testicular protein and cholesterol contents of albino rat

In this study the effect of thiazide diuretic compound on the protein and cholesterol contents in the testes of albino rats as the experimental model. The drug thiazide was administered orally daily for 10,20 and 30 days at the dose of 100mg/kg body weight. Total protein decrease in the testes of rats were evidenced may be due to the side effects of thiazide drug compound which is linked with the hyponatremia and protein metabolism. An elevated level of cholesterol contents observed in thiazide treated rats also revealed that the side effect of drug compound thiazide and also may be due to the stimulation of catecholamine which is stimulated therefore, the biochemical estimation such as protein and cholesterol in the testes after the thiazide treatment determined the effectiveness of diuretic drug compound would provide clinical evidences of their side effects.     

Acute and chronic toxicity studies on partially purified hypoglycemic preparation from water extract of bark ofFicus bengalensis

Acute and chronic toxicity studies were conducted to assess toxicity of a partially purified preparation from the water extract of the bark ofFicus bengalensis, which was demonstrated in our earlier studies to have significant hypoglycemic and hypocholesteroiemic effect on alloxan induced, mild and severe diabetes in rabbits. LD₅₀ of this preparation was found to be ∼1 gm/kg in rats when given orally. For chronic toxicity studies 3 doses of aqueous preparation were given to 3 groups of rats. First group received 5 times ED₅₀ (50 mg/kg), second group 10 times ED₅₀ (100 mg/kg) and the third group 15 times ED₅₀ (150 mg/kg) for 3 months. Fourth group which served as control was given water. After three months, blood was collected for studying biochemical and hematological parameters. Blood glucose, serum cholesterol, liver and kidney function tests, haemoglobin, total and differential leukocyte count were determined. Animals were sacrificed and histopathological examination of liver, heart and kidneys was carried out. Results of the study showed that partially purified preparation fromFicus bengalensis is not toxic by all the above mentioned parameters.     

Antidiabetic effect ofT. arjuna bark extract in alloxan induced diabetic rats

The present study was carried out to evaluate the antidiabetic effect of T. arjuna stembark extract and to study the activities of hexokinase, aldolase and phosphoglucoisomerase, and gluconeogenic enzymes such as glucose-6-phosphatase and fructose-1,6-diphosphatase in liver and kidney of normal and alloxan induced diabetic rats. Oral administration of ethanolic extract of bark (250 and 500mg/kg body weight) for 30 days, resulted in significant decrease of blood glucose from 302.67±22.35 to 82.50±04.72 and in a decrease in the activities of glucose-6-phosphatase, fructose-1,6-disphosphatase, aldolase and an increase in the activity of phosphoglucoisomerase and hexokinase in tissues. However, in the case of 250 mg/kg body weight of extract, less activity was observed. The study clearly shows that the bark extract ofT. arjuna possesses potent antidiabetic activity.     

Antioxidant activity of ethanol extract of rhizome ofPicrorhiza kurroa on indomethacin induced gastric ulcer during healing

Oral administration of ethanol extract of the rhizome ofPirorhiza kurroa at a dose of 20mg/kg body weight, for 10 consecutive days, was found to enhance the rate of healing on Indomethacin-induced gastric ulcer in rats, compared to the ulcerated group without treatment. The level of peroxidised lipid, in terms of thiobarbituric acid reactive species (TBARS), in gastric tissue, was increased in ulcerated rats which was restored to near normalcy on treatment with ethanol extract. The specific activity ofin vivo antioxidant enzymes, viz SOD and catalase and total tissue sulfhydryl (thiol) group, which were markedly decreased in ulcerated group, were found to be significantly elevated (p<0.05), on treatment with the above extract, at the specified dose, compared to the indomethacin—induced ulcerated group without any supporting treatment. The present study thus suggests that the ethanol extract of rhizome ofPicrorhiza kurroa, at the dose of 20mg/kg body weight, accelerated the healing of stomach wall of indomethacin induced gastric ulcerated rats by anin vivo free radical scavenging action.     

Alteration in some antioxidant enzymes in cardiac tissue upon monosodium glutamate [MSG] administration to adult male mice

4mg and 8mg monosodium glutamate per gram body weight was administered subcutaneously for 6 consecutive days to normal adult male mice and its effect was seen on 31^st day after the last injection on some antioxidant enzymes in heart. A significant dose dependent increase in lipid peroxidation and xanthine oxidase level was observed, whereas the activity of free radical scavenging enzymes such as superoxide dismutase and catalase was decreased in both monosodium glutamate treated groups (Group-2 and Group-3). So, the present work suggested that monosodium glutamate at dose level of 4mg/g body weight and above induced oxidative stress in the cardiac tissue by changing the activity of free radical initiating enzyme such as xanthine oxidase and scavenging enzymes like superoxide dismutase and catalase.     

Antidiabetic and ameliorative potential of Ficus bengalensis bark extract in streptozotocin induced diabetic rats

The aim of the present study was to evaluate the antidiabetic and ameliorative potential of aqueous extract of Ficus bengalensis bark in streptozotocin induced diabetic rats. The effect of oral administration of aqueous extract of F. bengalensis bark on blood glucose, serum electrolytes, serum glycolytic enzymes, liver microsomal protein, hepatic cytochrome P-450 dependent monooxygenase enzymes and lipid peroxidation in liver and kidney of streptozotocin -induced diabetic rats was studied. Oral administration of Ficus bengalensis to fed, fasted and glucose loaded diabetic rats significantly [F > 0.05 (ANOVA) and P< 0.05 (DMRT)] decreased the blood glucose level at 5 hrs and restored the levels of serum electrolytes, glycolytic enzymes and hepatic cytochrome P-450 dependent enzyme systems and decreased the formation of liver and kidney lipid peroxides at the end of 12 weeks. Further, the aqueous extract of Ficus bengalensis at a dose of 500mg/kg/day exhibits significant antidiabetic and ameliorative activity as evidenced by histological studies in normal and Ficus bengalensis treated streptozotocin induced diabetic rats. On the basis of our findings, it could be used as an antidiabetic and ameliorative agent for better management of diabetes mellitus.     

Evaluation of the radioprotective effect of turmeric extract and vitamin E in mice exposed to therapeutic dose of radioiodine

The aim of this study was to evaluate the radioprotective effect of turmeric extract (40 mg/kg body weight) and vitamin E (α- tocopherol acetate, 400 IU/kg body weight) supplementation on lipid peroxidation, reduced glutathione and antioxidant defense enzymes in various organs like liver, kidney and salivary glands at 24 h in adult Swiss mice. ¹³¹Iodine exposure significantly increased lipid peroxidation in kidney and salivary glands in comparison to control animals. Pre supplementation with turmeric extract for 15 days showed significant lowering of lipid peroxidation in kidney. On the other hand vitamin E pre supplementation showed marked reduction in lipid peroxidation in salivary glands. Reduced glutathione levels decreased significantly in liver after radiation exposure. However, pre supplementation with turmeric extract and vitamin E did not improve glutathione levels in liver. In conclusion, we have observed differential radioprotective effect of turmeric extract and vitamin E in kidney and salivary glands. However, Vitamin E seems to offer better radioprotection for salivary glands which is known to be the major site of cellular destruction after radioiodine therapy in patients.     

Effect of ethanolic leaf extract of <Emphasis Type="Italic">Trianthema portulacastrum</Emphasis> L. On aflatoxin induced hepatic damage in rats

The aim of this study was to investigate the ethanolic leaf extract of Trianthema portulacastrum L. (Family: Aizoaceae) on aflatoxin induced hepatic damage in rats. Aflatoxin intoxication in rats significantly (p < 0.001) elevated the levels of serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP), and total bilirubin, which indicated acute hepatocellular damage and biliary obstruction. Ethanolic leaf extract of T. portulacastrum showed dose dependent decrease in the levels of SGPT, SGOT, ALP and total bilirubin. Minimum effective dose of extract was found to be 100 mg/kg body weight. Results obtained from histopathological studies also supported hepatoprotective activity against aflatoxin-induced hepatotoxicity. Thus the study demonstrates that T. portulacastrum possess antihepatotoxic effect against aflatoxin.     

Additive Effect of Lipid Lowering Drug (Simvastatin) in Combination with Antidiabetic Drug (Glibenclamide) on Alloxan Induced Diabetic Rats with Long Term Dyslipidemia

High blood glucose level, elevated level of liver enzyme, necrosis and shrinkage of islets of Langerhans has been implicated in the pathogenesis of type 2 diabetes. High blood glucose cause oxidative stress, production of free radical as well as elevated SGPT and SGOT level. Both glibenclamide and simvastatin in fixed dose used as antihyperglycemic antidyslipidemic and antioxidative agents for type 2 diabetes treatment. This study therefore aimed to evaluate the antihyperglycemic, antidyslipidemic and antioxidative effect of fixed dose combination of glibenclamide (0.6 mg/70 kg body weight) and simvastatin (5 mg/70 kg body weight) on long term alloxan induced diabetic rats with cardiovascular disease using various diagnostic kits as a parameter of phamacotherapeutic and pharmacological effect. The study was carried out using 96 Swiss Albino male rats weighing about 200–220 g. Combination therapy induced a significant decrease in blood glucose level in alloxan induced diabetic rats, from 33.75 ± 1.65 to 5.80 ± 0.07 mmol/l 2 h after last dose administration, after 4 weeks treatment. In case of dyslipidemic effect, combination therapy reduced total cholesterol (45 %), triglyceride (36 %) and low density lipoprotein-cholesterol (32 %) levels significantly and increased high density lipoprotein-cholesterol level (57 %) in comparison with their respective diabetic control groups. Results of this study showed that combination therapy effectively decreased SGPT (ALAT) (55 %) and SGOT (ASAT) (51 %) in comparison with diabetic control group. It was also observed that catalase and superoxide dismutase enzyme activity was increased by 58 and 91 % respectively in comparison with diabetic control group after 4 weeks treatment with combination of both drugs. In conclusion, these findings of combination therapy (glibenclamide and simvastatin) on alloxan induced diabetes in rats are significantly better than monotherapy using single drug. The results of the present study suggest that, combination of the fixed dose of glibenclamide and simvastatin might be efficacious in patients with diabetic dyslipidemia and increased oxidative stress. Furthermore, this combination therapy offer dosage convenience to the patients and by virtue of its dual mode of action might be a useful addition to the therapeutic armamentarium for patients with diabetic dyslipidemia and oxidative stress.     

Hypolipidemic Activity of Cassia tora Seeds in Hyperlipidemic Rats

The hypolipidemic activity of Cassia tora (Chakvat, Chakunda) (Family: Caesalpiniaceae) seeds extract have been studied in two models of hyperlipidemia in rats. In an acute model, hyperlipidemia was induced by injecting a single dose of Triton WR-1339 (400 mg/kg, b.w.) intraperitonially in rats. Feeding with C. tora seed extract at the dose of 500 mg/kg, b.w. exerted significant lipid lowering effect as assessed by the reversal of plasma levels of total cholesterol, phospholipids, triglyceride and reactivation of post heparin lipolytic activity. In the chronic model, hyperlipidemia was induced by feeding with cholesterol rich-HFD in rats. The treatment with seeds extract of C. tora (500 mg/kg, b.w.) simultaneously for 15 days also caused lowering of lipid levels in plasma and liver following reactivation of plasma post heparin lipolytic activity and hepatic lipoprotein lipase activity in animals. The hypolipidemic activity of C. tora seeds was compared with a standard drug guggulipid (200 mg/kg, b.w.) in both models.     

Hepatoprotective and Nephroprotective Effect of Curcumin Against Copper Toxicity in Rats

Curcumin is a natural anti-inflammatory and antioxidant with several potential health benefits. Although it has been examined in several metals toxicity studies, but its role in the protection against copper toxicity has not been investigated. In this study; the detoxification and antioxidant effect of curcumin were examined to determine its prophylactic/therapeutic role experimentally in rats. Forty albino rats were divided into five groups; control, CuSO₄ (4 mg/kg body weight), curcumin (80 mg/kg body weight), curcumin post-treatment (CuSO₄ for 15 days followed by curcumin for the next 15 days) and curcumin co-treatment (CuSO₄ plus curcumin for 30 days). All rats were treated orally by stomach tube for 30 days/once a day. Changes were observed in hepatic marker enzymes such as: aspartate aminotransferase (AST), alanine transaminase-(ALT), alkaline phosphatase (ALP) and gamma-glutamyltransferase (GGT), besides the serum total protein, urea and creatinine. Concentration of liver and kidney antioxidants such as: catalase (CAT), superoxide dismutase (SOD), reduced glutathione-(GSH) and malondialdehyde (MDA) were measured. An increased in the activities of liver marker enzymes, urea, creatinine and the MDA contents were detected after exposure to CuSO₄. Meanwhile, the activities of serum total protein, hepatic and renal antioxidants were decreased. Changes in all biochemical parameters were alleviated by the post-treatment and co-treatment of curcumin. Our finding suggests that the curcumin showed protective effects on CuSO₄-induced hepatotoxicity and nephrotoxicity.     

Toxicology and free radicals scavenging property of Tamra bhasma

Free radicals are implicated in various chronic diseases. There has always been a search for new antioxidants. In this paper we have investigated Tamra bhasma, a metallic ayurvedic preparation. It is a time-tested medicine in Ayurveda and is in clinical use for various ailments specifically the free radical mediated diseases. Our results show that Tamra bhasma inhibits lipid peroxidation (LPO), prevents the rate of aerial oxidation of reduced glutathione (GSH) content and induces the activity of superoxide dismutase (SOD) in rat liver homogenate in the bi-phasic manner. The drug was orally given for 7, 15 and 30 days in different doses. Best protective response was found at the dose of 0.5mg/100g body weight in albino rats, although it showed some histopathological changes at the dose of 20mg/100g body weight. The results suggest that this Ayurvedic preparation is not merely a source of copper metal, but it is a strong anti-oxidant with no detectable adverse effect in lower doses of therapeutic range.     

Protective Effect of Emblica officinalis Against Alcohol-Induced Hepatic Injury by Ameliorating Oxidative Stress in Rats

The effect of Emblica officinalis fruit extract (EFE) against alcohol-induced hepatic damage in rats was investigated in the present study. In vitro studies showed that EFE possesses antioxidant as well nitric oxide (NO) scavenging activity. In vivo administration of alcohol (5 g/kg b.wt/day) for 60 days resulted increased liver lipid peroxidation, protein carbonyls, nitrite plus nitrate levels. Alcohol administration also significantly lowers the activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase and reduced glutathione as compared with control rats. Administration of EFE (250 mg/kg body weight) to alcoholic rats significantly brought the plasma enzymes towards near normal level and also significantly reduced the levels of lipid peroxidation, protein carbonyls and restored the enzymic and non-enzymatic antioxidants level. This observation was supplemented by histopathological examination in liver. Our data indicate that the tannoid, flavonoid and NO scavenging compounds present in EFE may offer protection against free radical mediated oxidative stress in rat hepatocytes of animals with alcohol-induced liver injury.     

Effect of water extract of <Emphasis Type="Italic">Trichosanthes dioica</Emphasis> fruits in streptozotocin induced diabetic rats

In rats with streptozotocin induced severe diabetes mellitus, aqueous extract of Trichosanthes dioica fruits at a dose of 1000mg/kg body weight daily once for 28 days reduced the levels of fasting blood glucose, postprandial glucose, asparate amino transferase, alanine amino transferase, alkaline phosphatase, creatinine, urine sugar and urine protein where as total protein and body weight was increased. No toxic effect was observed during LD50. Our study suggests that further detailed toxicity studies and mechanism of action of T. dioica would be useful for undertaking human trials.     

Antioxidant activity ofcurculigo orchioides in carbon tetrachloride—induced hepatopathy in rats

In this study antioxidant activity of methanol extract of rhizomes ofCurculigo orchioides (MEC) was investigated using carbon tetrachloride (CCl₄)-intoxicated rat liver as the experimental model. The hepatotoxic rats were administered MEC for 90 days (daily, orally at the dose of 70 mg per kg body weight). Lipid peroxidation (LPO) in CCl₄-intoxicated rats was evidenced by a marked increment in the levels of thiobarbituric acid reactive substances (TBARS) and diene conjugates (CD), and also a distinct diminution in glutathione (GSH) content in the liver. In CCl₄+MEC—treated rats these biochemical parameters attained an almost normal level. The decreased activity of antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and glutathione reductase (GRD) in CCl₄—intoxicated rats, and its retrieval towards near normalcy in CCl₄+MEC—administered rats revealed the efficacy of MEC in combating oxidative stress due to hepatic damage. Elevated level of glutathione transferase(GTS) observed in hepatotoxic rats too showed signs of retuming towards normalcy in MEC co-administered animals, thus corroborating the antioxidant efficacy of MEC. The findings provide a rationale for further studies on isolation of active principles and its pharmacological evaluation.     

Hypoglycemic and hypolipidemic effects of diacetodibutyl disulphide

The hyperlipidemia, fatty liver and the high levels of liver and kidney thiobarbituric acid reactive substances (TBARS) observed in rats which were fed ethanol for 45 days, could be significantly reduced by feeding diacetodibutyl disulphide (DADBDS). Ethanol-induced hypoproteinemia and the rise in serum enzymes like AST (EC 2.6.1.10), ALT (EC 2.6.1.2) and ALP (EC 3.1.3.1) could also be ameliorated by DADBDS. Feeding of this compound to normal rats did not produce any change in serum or tissue lipid levels or serum enzymes or tissue TBARS except a moderate reduction in serum triacyl glycerols. DADBDS feeding to rats maintained on a high lipid diet could also reduce the serum and tissue lipid levels and also reduce the serum transaminases. DADBDS which is an aliphatic disulphide could produce hypolipidemic effects in rats fed a single large dose of ethanol, whereas dimenthol disulphide which is an aromatic disulphide was not useful as a hypolipidemic agent. Perhaps hypolipidemic effects are shown only by aliphatic disulphides and not by aromatic disulphides. Feeding of 100 mg DADBDS per kg body weight to normal fasted rats produced a mild hypoglycemia, but higher doses produced a hyperglycemic effect. This dose of DADBDS increased the serum insulin levels and reduced blood glucose levels in fasted diabetic rats, but DADBDS feeding did not alter the serum insulin levels in fasted normal rats. DADBDS is odourless and tasteless in 1% solution and it could be a better substitute for garlic for hypoglycemic and hypolipidemic studies.     

Biochemical activity of selenium and glutathione on country made liquor (CML) induced hepatic damage in rats

The serum and hepatic enzymes of rats were studied after exposed to country made liquor (CML) along with two chelating agents (glutathione and Selenium). There was a significant increase in several serum enzyme levels (viz., aspartate transaminase, alanine transaminase, alkaline phosphatase, sorbitol dehydrogenase, glutamate dehydrogenase, bilirubin) and decrease in various hepatic enzymes (Succinic dehydrogenase, Glucose 6-phosphatase, 5'Nucleotiease, Acid phosphatase, Acid ribonuclease, Cytochrome P-450) due to repeated administration of CML (2ml/100g of body weight). Results of this study revealed that the GSH and Se could give a significant protective action in serum and hepatic enzymes of CML exposed rats.     

Effects of chronic ethanol exposure on renal function tests and oxidative stress in kidney

After administration, ethanol and its metabolites go through the kidneys and are excreted into urine. The kidney seems to be the only vital organ generally spared in chronic alcoholics. Therefore, we investigated the multiple effects of chronic ethanol exposure on renal function tests and on oxidative stress related parameters in the kidney. Chronic ethanol (1.6 g ethanol/ kg body weight/ day) exposure did not show any significant change in relative weight (g/ 100g body weight) of kidneys, serum calcium level or glutathione s-transferase activity. However, urea and creatinine concentration in serum, and TBARS level in kidney elevated significantly, while reduced glutathione content and activities of glutathione peroxidase, glutathione reductase and superoxide dismutase diminished significantly after 12 weeks of ethanol exposure. Catalase activity showed increased activity after 4 weeks of ethanol exposure and decreased activity after 12 weeks of ethanol exposure. Genesis of renal ultrastructural abnormalities after 12 weeks of ethanol exposure may be important for the development of functional disturbances. This study revealed that chronic ethanol exposure for longer duration is associated with deleterious effects in the kidney.