Prominent Pancreatic Lipase Inhibition and Free Radical Scavenging Activity of a Myristica fragrans Ethanolic Extract in vitro. Potential Role in Obesity Treatment.

Objective:The objective of the present study was to evaluate the antioxidant and lipase inhibitory potential of various extracts of Myristica fragrans (in vitro). Material and methods:Ethanolic extracts of Myristica fragrans were studied for their free radical scavenging and lipase inhibitory potentials by using porcine lipase, PNPB and DPPH. All results were obtained by applying active formulas and calculating the percentage of inhibition. Results:Among all extracts, Myristica fragrans ethanolic extract has shown the strongest pancreatic lipase inhibitory activity at 100 ìg/mL (66.24%), with the closest potency to tthat of the standard drug, Orlistat (81.57%). This extract has also exhibited a potent antioxidant activity. The findings of the present study clearly showed that DPPH free radical scavenging activity of MFE produced 88% inhibition at 5 mg/mL as compared to standard ascorbic acid, which was 90%. Conclusions:Ethanolic extracts of Myristica fragrans had a marked PL inhibitory action and antioxidant effect. Therefore, based on this research evidence, they could be aternatively used for obesity treatment.

Herbal formulation, DIA-2 and Rosiglitazone ameliorates hyperglycemia and hepatic steatosis in type 2 diabetic rats.

DIA-2 is a herbal mixture containing standardized extract of Allium sativum and Lagerstroemia speciosa. Recently we have reported the anti-diabetic effect of DIA-2 in high fat diet (HFD) and streptozotocin (STZ) induced type 2 diabetic (T2D) rats. The purpose of this study was to investigate and compare the effects of DIA-2 with Rosiglitazone (RG) on plasma biomarkers of hepatocellular injury, liver carbohydrate metabolizing enzymes, glycogen content, oxidant/antioxidant status and histopathological changes in T2D rats. ALT and ALP levels were significantly decreased after DIA-2 and RG treatment compared to T2D rats. Total protein and albumin remained unaltered in all the groups. Significant decrease in AST levels were observed after DIA-2 (125 mg/kg) and RG treatment. Hepatic hexokinase activity was significantly increased after RG and DIA-2 treatment and fructose-1, 6-bisphosphatase activity were inversely correlated with hexokinase activity. Hepatic gucose-6-phosphatase activity was significantly (p < 0.05) reduced after DIA-2 (62.5 mg/kg) and RG treatment. Lipid peroxides levels was significantly decreased in the liver of DIA-2 (62.5; p < 0.01 & 125 mg/kg; p < 0.05) treated animals. Hepatic glycogen content (p < 0.05) and antioxidant enzymes [SOD (p < 0.01; 62.5 mg/kg); GPx and GSH (125 mg/kg; p < 0.01)] were significantly increased after DIA-2 treatment. RG treatment on hepatic glycogen, GPx (p < 0.01) and SOD, GSH (p < 0.05) levels were significant when compared to T2D rats. These biochemical parameters were also correlated with histopathological evaluation. The above findings revealed that administration of DIA-2 could ameliorate the biochemical and histopathological changes in liver of T2D rats indicating the protective role of DIA-2 against HFD/STZ induced diabetes. In addition, DIA-2 and RG treatment resulted in amelioration of hepatic steatosis in T2D rats.

Antioxidants and tumor necrosis factor alpha-inhibiting activity of sesame oil against doxorubicin-induced cardiotoxicity.

OBJECTIVE: Oxidative stress is currently considered to be the key factor in doxorubicin-induced cardiotoxicity. Comparatively small quantity of the endogenous antioxidant content of the heart is assumed to be the predisposing factor for doxorubicin-induced cardiotoxicity. The present research was designed to evaluate the antioxidant potential and tumor necrosis factor alpha-(TNF-α) inhibiting activity of sesame oil against acute doxorubicin-induced cardiotoxicity. METHODS: Male Wistar albino rats (180-200 g) were administered sesame oil in two dissimilar doses (5 and 10 ml/kg body weight, orally) for 30 days, followed by a single dose of doxorubicin (30 mg/kg s.c.). RESULTS: In the doxorubicin-treated group, increased oxidative stress was proven by a significant rise of thiobarbituric acid reactive substances level and a decrease of myocardial superoxide dismutase, catalase and reduced glutathione content. Histopathological studies showed myocardial necrosis with accumulation of inflammatory cells, vacuolization and overall enlargement of the myocardium. Western blot analysis showed marked expression of TNF-α in the myocardium. Alteration in biochemical parameters by doxorubicin administration was prevented significantly (p < 0.0001) in the 5 and 10 ml/kg sesame oil treated rat hearts. Treatment with 5 and 10 ml/kg of sesame oil reduced the doxorubicin-induced TNF-α expression in the myocardium, which was associated with reduced myocyte injury. The overall effect of sesame oil was comparable with probucol, which shows similar protection. CONCLUSION: The chronic oral administration of sesame oil prevents acute doxorubicin-induced cardiotoxicity by enhancing cardiac endogenous antioxidants and decreasing myocardial TNF-α expression.

DIA-2, a polyherbal formulation ameliorates hyperglycemia and protein-oxidation without increasing the body weight in type II diabetic rats.

BACKGROUND: The dried bulbs of Allium sativum (Garlic) and leaves of Lagerstroemia speciosa (Banaba) are used as medicinal food for the treatment of diabetes and other ailments. AIM: The present study was undertaken to ascertain whether the combination of both garlic and banaba extract produces synergistic therapeutic effect in diabetic state. METHODS: In the in vitro studies, the effect of standardized aqueous extract of Allium sativum (ASE), methanolic extract of Lagerstroemia speciosa (LSE) and their mixture (1:1 ratio), DIA-2 on insulin stimulated glucose uptake in 3T3-L1 cells, erythrocyte sorbitol accumulation and protein glycation were evaluated. Impetus from the in vitro findings triggered to screen the anti-diabetic potential of DIA-2 in rat model of type II diabetes and associated oxidative stress. In the in vivo studies, acute oral toxicity of DIA-2 was determined following OECD-423 guidelines in female rats. Anti-diabetic activity of DIA-2 was investigated in high fat diet/low dose streptozotocin induced type II diabetes at four dose levels (62.5, 125, 250 and 500 mg/kg b.w) in rats. RESULTS: Combination of ASE and LSE produced synergistic and a dose dependent increase in glucose uptake in 3T3 adipocyte cell lines when compared to the individual extracts. A similar effect was observed in the inhibition of sorbitol accumulation and protein glycation tests. DIA-2 restored the glucose and lipid level near to normal level without gain in body weight which is the most commonly encountered side effect with the use of conventional antidiabetic agents, particularly insulin, insulin secretagogues, sulfonylureas and thiazolidinediones. DIA-2 also decreased hepatic protein carbonyl content levels significantly in the diabetic rats. CONCLUSIONS: The study concluded that DIA-2 posses potent anti-diabetic activity and anti-oxidant effects.  

In vitro cytotoxic, antioxidative and alpha-glucosidase inhibitory potential of a herbal mixture comprised of Allium sativum and Lagerstroemia speciosa.

BACKGROUND: Hyperglycemia induced over production of free radicals in the mitochondrial electron transport chain is now considered as one of the central mechanisms in the pathogenesis of diabetic complications. Allium sativum and Lagerstroemia speciosa contains active principles possessing anti-diabetic and antioxidant properties. This study is aimed at evaluating the evidence that supports this traditional claim and investigates the possible synergistic effect on these herbs when given as a herbal mixture in vitro. AIM: The present study investigates the cytotoxic, antioxidant and a-glucosidase inhibitory potential of Allium sativum (ASE), Lagerstroemia speciosa (LSE) and their combinations using in vitro methods. MATERIALS AND METHODS: The total phenol, total flavonoid and total tannin content were determined in ASE and LSE. The cytotoxic effects of ASE, LSE and their combination in the ratio of 1:2, 1:1 w/w were evaluated using 3T3 L1 preadipocyte cells. Effect of ASE, LSE and its mixture on intracellular reactive oxygen species (ROS) production were determined by 2', 7'dichlorfluorescein diacetate (DCF DA) staining technique in 3T3-L1 adipocytes. The ability of the herbal extracts and their combination to scavenge super oxide radicals and to inhibit alpha-glucosidase enzyme (a carbohydrate metabolising enzyme) were measured using in vitro methods. RESULTS: The total phenols and tannins were expressed as microgram (microg) of gallic acid equivalents/mg of extract (GAE/mg), flavonoids as microg of quercetin equivalents/mg of extract (QE/mg). LSE had significant higher total phenol (300.11 +/- 1.99), flavonoid (53.12 +/- 0.48) and tannin content (118.90 +/- 0.15) compared to ASE which possessed total phenol (159.93 +/- 0.87); flavonoid (9.37 +/- 0.73) and tannin content (80.5 +/- 0.19). The IC50 value, the concentration of the extracts that cause 50% inhibition or cell death was measured as an index of cytotoxicity. The IC50 value was found to be in the following decreasing order: 1:2 mixture (98 microg/ml) > ASE (323.6 microg/ml) > 1:1 mixture (428.1 microg/ml) > LSE (2154 microg/ml). The 1:1 mixture was comparatively less cytotoxic under the tested concentration range (1 x 10(0) pg - 1 x 10(8) pg) than 1:2 combinations. The results observed with lactate dehydrogenase (LDH) release were similar to that of cell viability assay. The 1:1 mixture (DIA-2 hereafter) was considered for further investigations. DIA-2 inhibited the ROS levels, which is evidenced by the decreased DCF fluorescence. DIA-2 could also efficiently scavenge the super oxide radical generated from PMS/NADH-NBT system showing an IC50 value 69.99 microg/ml, the IC50 value of ASE (157.7 microg/ml), LSE (20.43 microg/ml), and ascorbic acid (49.64 microg/ml) used as positive control. The results of in vitro a-glucosidase inhibitory assay showed highest IC50 value with LSE (0.3 microg/ml) and DIA-2 (0.7 microg/ml) than ASE (136.3 microg/ml) and positive control miglitol (651.8 microg/ml). CONCLUSIONS: DIA-2 exerts synergistic effect in scavenging the ROS and inhibiting the enzyme alpha-glucosidase in vitro compared to its individual extracts. The possible synergistic therapeutic effects may be due the presence of the antioxidant rich flavonoids, phenols and tannins present in LSE and ASE.

Floral extract of Tecoma stans: a potent inhibitor of gentamicin-induced nephrotoxicity in vivo.

OBJECTIVE: To highlight the nephroprotective activity of ethyl acetate extract of dried flowers of Tecoma stans for its protective effects on gentamicin-induced nephrotoxicity in albino rats. METHODS: For studying acute toxicity study, single oral dose of 5,000 mg ethyl acetate floral extract/kg body weight was administered to albino rats (five females, five males). Nephrotoxicity was induced in albino rats by intraperitoneal administration of gentamicin 80 mg/kg/day for eight days. Effect of concurrent administration of ethyl acetate floral extract of Tecoma stans at a dose of 100, 200 and 300 mg/kg/day given by oral route was determined using serum creatinine, serum uric acid, blood urea nitrogen and serum urea as indicators of kidney damage. The study groups contained six rats in each group. As nephrotoxicity of gentamicin is known to involve induction of oxidative stress, in vitro antioxidant activity and free radical-scavenging activity of this extract was also evaluated. RESULTS: For acute toxicity testing both female and male rats administered with the extract at a dose of 5,000 mg/kg. The results showed no toxicity in terms of general behavior change, mortality, or change in gross appearance of internal organs (LD(50) > 5 000 mg/kg). It was observed that the ethyl acetate floral extract of Tecoma stans significantly protected rat kidneys from gentamicin-induced histopathological changes. Gentamicin-induced glomerular congestion, peritubular and blood vessel congestion, epithelial desquamation, accumulation of inflammatory cells and necrosis of the kidney cells were found to be reduced in the groups receiving the ethyl acetate floral extract of Tecoma stans along with gentamicin in a dose dependent manner. The floral extract also reduced the gentamicin-induced increase in serum creatinine, serum uric acid, blood urea nitrogen and serum urea levels (P >0.01). CONCLUSIONS: The present study indicates a very important role of reactive oxygen species (ROS) and the relation to renal dysfunction and point to the therapeutic potential of Tecoma stans in gentamicin induced nephrotoxicity.

Anti-ulcer activity of crude alcoholic extract of Toona ciliata Roemer (heart wood).

The ethanol extract of Toona ciliata Roemer (heart wood) was evaluated for its anti-ulcer activity against aspirin plus pylorous ligation induced gastric ulcer (antisecretory), HCl-ethanol induced ulcer (cytoprotective) and water immersion stress induced ulcer in rats. We found that Toona ciliata extract at a dose of 300mg/kg p.o. markedly decrease the incidence of ulcers in all the three models. Ethanol extract of Toona ciliata showed significant reduction in gastric volume, free acidity, total acidity and ulcer index. The plant extract also showed gastro protective activity (52.94%), whereas standard drug sucralfate showed 94.85%. Toona ciliata extract showed protection index 43.0% in water immersion stress induced ulcer, whereas standard drug omeprazole showed protection index 100%.

Protective effect of Indigofera aspalathoides against CCl4-induced hepatic damage in rats.

The alcoholic extract of stem of Indigofera aspalathoides was evaluated for its antihepatotoxic activity against CCl(4)-induced hepatic damage in rats. The activity was evaluated by using biochemical parameters, such as serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP), total bilirubin and gama glutamate transpeptidase (GGTP). The histopathological changes of liver sample were compared with respective control. The extract showed remarkable hepatoprotective effect.

Effect of methanolic extract of Enicostemma littorale on Dalton's ascitic lymphoma.

The antitumour activity of methanolic extract of Enicostemma littorale (MEL) has been evaluated against Dalton's ascitic lymphoma (DAL) in swiss albino mice. A significant enhancement of mean survival time of MEL treated tumour bearing mice was found with respect to control group. MEL treatment was found to enhance peritoneal cell counts. When these MEL treated animals underwent i.p. inoculation with DAL cells, tumour cell growth was found to be inhibited. After 14 days of inoculation, MEL is able to reverse the changes in the haemotological parameters, protein and PCV consequent to tumour inoculation.

Antitumour activity of rhinacanthone against Dalton's ascitic lymphoma.

The antitumour activity of Rhinacanthone (3,4-dihydro-3,3-dimethyl-2H-naphtho-[1,2-B] pyran-5,6-dione) has been evaluated against Dalton's ascitic lymphoma (DAL) in Swiss albino mice. A significant enhancement of mean survival time of tumour bearing mice and peritoneal cell count in normal mice was observed with respect to the control group. When these Rhinacanthone treated animals underwent i.p. inoculation with DAL cells, tumour cell growth was found to be inhibited. After 14 d of inoculation, Rhinacanthone was able to reverse the changes in the haemotological parameters, protein and packed cellular volume consequent to tumour inoculation.

Effect of the methanolic extract of Glinus lotoides on Dalton's ascitic lymphoma.

The antitumour activity of the methanolic extract of Glinus lotoides (MGL) has been evaluated against Dalton's ascitic lymphoma (DAL) in Swiss albino mice. A significant enhancement of mean survival time of tumour bearing mice and peritoneal cell count in normal mice was observed with respect to the control group. When these MGL treated animals underwent i.p. inoculation with DAL cells, tumour cell growth was found to be inhibited. After 14 d of inoculation, MGL is able to reverse the changes in the haemotological parameters, protein and packed cellular volume consequent to tumour inoculation.

Anti-steroidogenic activity of floral extract of Thespesia populnea Corr. in mouse ovary.

Anti-steroidogenic activity of various extracts of T. populnea was screened in female albino mice. The weight of the uterus and ovaries were reduced significantly and the cholesterol and ascorbic acid content in ovaries were significantly elevated due to the treatment with extract of T. populnea. The significant inhibition of delta 5, 3 beta hydroxy steroid dehydrogenase and glucose-6-phosphate dehydrogenase, the two key enzymes involved in ovarian steroidogenesis were also observed in mouse ovaries after 15 days of treatment.

Anti - inflammatory and antimicrobial activities of the root, bark and leaves of azadirachta indica.

Azadirachta indica is a plant of varied uses in Ayurveda since ancient times and is highly extolled by expert physicians and as well as practitioners of folk medicines. Almost every part of the tree has long been used in folklore and traditional systems of medicine for the treatment of a variety of human ailments. The 50% acetone extract of the root, bark and leaves of A. indica sowed marked anti- inflammatory activity in carrageenan induced edema in rats, Antimicrobial activity was also tested.