Evaluation of effect of alcoholic extract of heartwood of Pterocarpus marsupium on in vitro antioxidant, anti-glycation, sorbitol accumulation and inhibition of aldose reductase activity.

Rising popularity of phytomedicines in various diseased conditions have strengthened the significance of plant-research and evaluation of phytoextracts in clinical manifestations. Pterocarpus marsupium Roxb., a medicinal plant, known for its anti-oxidant and anti-diabetic activity is a rich source of phytochemicals with antihyperglycemic and antihyperlipidemic activities. However, its possible role in diabetic complications is not evaluated yet. The present study explores the possible role of alcoholic extract of heartwood of P. marsupium in the treatment of long-term diabetic complications. The alcoholic extract of P. marsupium was evaluated for advanced glycation-end-products formation, erythrocyte sorbitol accumulation and rat kidney aldose reductase enzyme inhibition at the concentration of 25-400 µg/ml using in-vitro bioassays. Also the phytoextract at the concentration of 10-320 µg/ml was evaluated for its antioxidant potential by in-vitro antioxidant assays which includes, determination of total phenol content; reducing power assay; nitric oxide scavenging activity; superoxide radical scavenging activity; total antioxidant capacity; total flavonoid content; DPPH scavenging activity; and hydrogen peroxide scavenging activity. The alcoholic extract of P. marsupium across varying concentrations showed inhibitory effect as evident by IC50 on advanced glycation-end-products formation (55.39 µg/ml), sorbitol accumulation (151.00 µg/ml) and rat kidney aldose reductase (195.88 µg/ml). The phytoextract also exhibited high phenolic and flavonoid contents with promising antioxidant potential against the antioxidant assays evaluated. The present investigation suggests that the phytoextract showed prominent antioxidant, antiglycation property and, inhibited accumulation of sorbitol and ALR enzyme, thus promising a beneficial role in reducing/delaying diabetic complications.

Comparative and quantitative analysis of antioxidant and scavenging potential of Indigofera tinctoria Linn. extracts.

OBJECTIVE: To compare and elucidate the antioxidant efficacy of ethanolic and hydroethanolic extracts of Indigofera tinctoria Linn. (Fabaceae family). METHODS: Various in-vitro antioxidant assays and free radical-scavenging assays were done. Quantitative measurements of various phytoconstituents, reductive abilities and chelating potential were carried out along with standard compounds. Half inhibitory concentration (IC50) values for ethanol and hydroethanol extracts were analyzed and compared with respective standards. RESULTS: Hydroethanolic extracts showed considerably more potent antioxidant activity in comparison to ethanol extracts. Hydroethanolic extracts had lower IC50 values than ethanol extracts in the case of DPPH, metal chelation and hydroxyl radical-scavenging capacity (829, 659 and 26.7 µg/mL) but had slightly higher values than ethanol in case of SO2- and NO2-scavenging activity (P<0.001 vs standard). Quantitative measurements also showed that the abundance of phenolic and flavonoid bioactive phytoconstituents were significantly (P<0.001) greater in hydroethanol extracts (212.920 and 149.770 mg GAE and rutin/g of plant extract respectively) than in ethanol extracts (211.691 and 132.603 mg GAE and rutin/g of plant extract respectively). Karl Pearson's correlation analysis (r2) between various antioxidant parameters and bioactive components also associated the antioxidant potential of I. tinctoria with various phytoconstituents, especially phenolics, flavonoids, saponins and tannins. CONCLUSION: This study may be helpful to draw the attention of researchers towards the hydroethanol extracts of I. tinctoria, which has a high yield, and great prospects in herbal industries to produce inexpensive and powerful herbal products.

Physicochemical and Antioxidant Assays of Methanol and Hydromethanol Extract of Ariel Parts of Indigofera tinctoria Linn.

Free radicals or reactive oxygen indices give rise to oxidative injury, which is a fundamental mechanism underlying a number of disease like diabetes, cancer, neurodegenerative disorders. Deleterious effects of reactive oxygen species can be nullified by using different natural antioxidants derived from plants Indigofera tinctoria is such plant. This study was planned in order to trace and determine the antioxidant capability of Indigofera tinctoria. All the reagents and chemicals used in this study were obtained from reliable firms. The plant extracts were subjected to phytochemical screening, quantitative assays and antioxidant profiling. The results suggested that plant extracts contained all pharmacologically important phytoconstituents in appreciable amounts and having excellent antioxidant capabilities.

Proteomic identification of Syzygium cumini seed extracts by MALDI-TOF/MS.

Syzygium cumini is traditionally used medicinal plant. The different part of the plant such as bark, leaves, seed and fruits are widely used as an alternative medicine in various diseases. Although the scientific community has a strong interest on S. cumini seed biochemistry focusing on metabolite composition, proteins have not yet been investigated. In the present study, we have applied a proteomic approach to study the proteome of the S. cumini seed using phenol extraction method for protein isolation, which were never analysed before. Fifteen brightly silver stained protein spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after resolving on two-dimensional gel electrophoresis. These proteins have been found to involve in various functions such as antifungal, sulphur metabolism, carbohydrate metabolism, fruit ripening and softening, dormancy breaking and seed germination, hormone signalling, secondary metabolite transport, defence and stress response, nitrogen metabolism, synthesis and stabilization. Amongst the identified protein, lactoferrin was a mammalian origin protein with high nutritious and pharmaceutical value, which was purified by different types of chromatographic techniques and confirmed by western blotting. The antibacterial activity of lactoferrin was assessed by disc diffusion assay. We suggest that the protein constituents of S. cumini may have role in various functions required for plant physiology and its dietary values.

Impact of gonadotropin supplementation on the expression of germ cell marker genes (MATER, ZAR1, GDF9, and BMP15) during in vitro maturation of buffalo (Bubalus bubalis) oocyte.

The present study was designed to investigate whether gonadotropins [follicle-stimulating hormone (FSH) and luteinizing hormone (LH)] and buffalo follicular fluid (bFF) supplementation in maturation medium influences the transcript abundance of germ cell marker genes [maternal antigen that embryos require (MATER), Zygote arrest 1 (ZAR1), growth differentiation factor 9 (GDF9), and bone morphogenetic protein 15 (BMP15)] mRNA in buffalo (Bubalus bubalis) oocytes. Buffalo ovaries were collected from local abattoir, oocytes were aspirated from antral follicles (5-8 mm) and matured in vitro using two different maturation regimens, viz, group A: gonadotropin (FSH and LH) and group B: non-gonadotropin-supplemented maturation medium containing 20% buffalo follicular fluid (bFF). mRNA was isolated from immature (330) and in vitro matured oocytes from both the groups (A, 320; B, 340), and reverse transcribed using Moloney murine leukemia virus reverse transcriptase. Expression levels of MATER, ZAR1, GDF9, and BMP15 mRNA transcripts were analyzed in oocytes of both maturation groups as well as immature oocytes using real-time PCR. QPCR results showed that GDF9 and BMP15 transcripts were significantly (p<0.05) influenced with gonadotropins and bFF supplementation during in vitro maturation of buffalo oocyte; however, MATER and ZAR1 transcripts were not influenced with gonadotropins and bFF supplementation in vitro. These results indicated that the expression levels of MATER, ZAR1, GDF9, and BMP15 mRNA were varied differentially during in vitro maturation of buffalo oocyte and were found to be gonadotropins (FSH and LH) or bFF dependent for GDF9 and BMP15.

Antihepatotoxic activity of Euphorbia neriifolia extract against N-nitrosodiethylamine-induced hepatocarcinogenesis in mice.

OBJECTIVE: To scrutinize the protective role of hydroethanolic extract of Euphorbia neriifolia leaves (HEEN) against N-nitrosodiethylamine (DENA)-induced hepatocarcinogenesis in male Swiss albino mice. METHODS: Experimental mice were pretreated with 150 and 400 mg/kg of HEEN, or 0.5% and 1% of butylated hydroxyanisole (BHA) as a standard for 14 d prior to the administration of a single dose of 50 mg/kg of DENA. Levels of xenobiotic metabolizing enzymes such as cytochrome (Cyt) P450 and Cyt b5, activities of reduced glutathione (GSH), glutathione-S-transferase (GST), aspartate amino transaminase (AST), alanine amino transaminase (ALT) and alkaline phosphatase (ALP), and total protein and cholesterol content in the liver tissues were measured to determine the hepatotoxicity caused by DENA. RESULTS: The levels of Cyt P450 and Cyt b5 were significantly increased, and GST and GSH were significantly depleted after DENA treatment (P<0.01). The activities of AST, ALT and ALP, and the total protein content were also significantly dropped off (P<0.01). The total cholesterol level was markedly increased by DENA as compared with the normal group (P<0.01). However, the pre-supplementation of HEEN showed a remarkable amelioration of these abnormalities, and the levels of the antioxidant enzymes in the liver were significantly restored, which exhibited the dose-dependent protective effect against DENA-induced hepatotoxicity. CONCLUSION: HEEN exerts its chemopreventive effects by alleviating the xenobiotic enzymes and enhancing the levels of antioxidants and biochemical assays in DENA-induced carcinogenesis by reducing the formation of free radicals.

Renoprotective effects of Moringa oleifera pods in 7,12-dimethylbenz [a] anthracene-exposed mice.

OBJECTIVE: To investigate the potential of hydroethanolic extract of Moringa oleifera (MOHE) against 7, 12-dimethylbenz [a] anthracene (DMBA)-induced toxicity in male Swiss albino mice. METHODS: Experimental mice were respectively pretreated with 200 and 400 mg/kg of MOHE, and 0.5% and 1% of butylated hydroxyanisole (BHA) for two weeks prior to the administration of 15 mg/kg of DMBA, respectively. Levels of xenobiotic metabolizing enzymes such as cytochrome (Cyt) P450 and Cyt b5, activities of reduced glutathione (GSH) and glutathione-S-transferase (GST) and renal aspartate amino transaminase (AST), alanine amino transaminase (ALT) and alkaline phosphatase (ALP), and content of protein and total cholesterol were measured to determine the nephrotoxicity caused by DMBA and to elucidate the ameliorating role of M. oleifera. RESULTS: Single oral administration of 15 mg/kg of DMBA resulted in significant increases in Cyt P450 and Cyt b5 (P<0.01). The toxic effect of DMBA was justified by the significant decreases in the activities of GSH and GST in renal tissues (P<0.05). The levels of renal AST, ALT and ALP and protein content which are indicative of renocellular damage were also found decreased along with significant increase in total cholesterol content in DMBA-treated mice (P<0.01). The DMBA-induced alterations in the tissues were significantly reversed after pretreatment with 200 and 400 mg/kg of MOHE orally for 14 d (P<0.01). CONCLUSION: The effects of MOHE in enhancing the levels of antioxidants and enhancing the levels of biochemical assays in DMBA-induced carcinogenesis are by reducing the formation of free radicals. This study rationalizes the ethnomedicinal use of M. oleifera for the protection against nephrotoxicity induced by chemical carcinogens.

Chemopreventive efficacy of Moringa oleifera pods against 7, 12-dimethylbenz[a]anthracene induced hepatic carcinogenesis in mice.

Oxidative stress is a common mechanism contributing to initiation and progression of hepatic damage in a variety of liver disorders. Hence there is a great demand for the development of agents with potent antioxidant effect. The aim of the present investigation is to evaluate the efficacy of Moringa oleifera as a hepatoprotective and an antioxidant against 7, 12-dimethylbenz[a]anthracene induced hepatocellular damage. Single oral administration of DMBA (15 mg/kg) to mice resulted in significantly (p<0.001) depleted levels of xenobiotic enzymes like, cytochrome P450 and b5. DMBA induced oxidative stress was confirmed by decreased levels of reduced glutathione (GSH) and glutathione-S-transferase (GST) in the liver tissue. The status of hepatic aspartate transaminase (AST), alanine transaminase (ALT), and alkaline phosphatase (ALP) which is indicative of hepatocellular damage were also found to be decreased in DMBA administered mice. Pretreatment with the Moringa oleifera (200 and 400 mg/kg) orally for 14 days significantly reversed the DMBA induced alterations in the liver tissue and offered almost complete protection. The results from the present study indicate that Moringa oleifera exhibits good hepatoprotective and antioxidant potential against DMBA induced hepatocellular damage in mice that might be due to decreased free radical generation.

Protective effect of Withania somnifera roots extract on hematoserological profiles against lead nitrate-induced toxicity in mice.

The in vivo protective role of hydro-methanolic root extract of Withania somnifera (WS) was evaluated in alleviating lead nitrate (LN)-induced toxicity in male Swiss albino mice by measuring hematoserological profiles. The lead-treated (20 mg/kg body wt, p.o.) albino mice (25-30 g) concurrently received the root extract (200 and 500 mg/kg body wt, p.o.) once daily for the duration of six weeks. Animals exposed to LN showed significant (P < 0.001) decline in haemoglobin content, red blood cell count, white blood cell count, packed cell volume and insignificant decrease in mean corpuscular haemoglobin and mean corpuscular haemoglobin content, while mean corpuscular volume and platelet count were increased. A significant elevation (P < 0.001) in serum glutamate oxaloacetate transaminase, serum glutamate pyruvate transaminase, alkaline phosphatase, acid phosphatase and total cholesterol were also observed, when compared with control mice. Thus, the study demonstrated that the concurrent daily administration of root extract of WS protected the adverse effects of LN intoxication in mice.

Chemopreventive effect of hydroethanolic extract of Euphorbia neriifolia leaves against DENA-induced renal carcinogenesis in mice.

The present study was conducted to investigate the chemopreventive effects of hydro-ethanolic extract of Euphorbia neriifolia (EN) on N-nitrosodiethylamine (DENA) induced renal cancer in male Swiss albino mice. Animals were pretreated with EN extract (150 and 400 mg/kg body weight; p.o) and butylated hydroxyanisole (BHA) as a standard (0.5% and 1% BHA p.o) both for two week prior to the administration of single dose of DENA (50 mg/kg body weight; p.o). Various in vivo antioxidant biochemical parameters like lipid peroxidation (LPO), superoxide dismutase (SOD), and catalase (CAT) were evaluated to determine the reno-protective and antioxidant activity of EN. DENA increased oxidative stress through increase in LPO and decrease in antioxidant enzymes (SOD, and CAT). The EN extract significantly restored the antioxidant enzyme level in the kidney and exhibited significant dose dependant protective effect against DENA induced nephrotoxicity, which can be mainly attributed to the antioxidant property of the extract. This study rationalized the ethno-medicinal use of EN for protection against renal cancer.

The effect of oral administration of Allium sativum extracts on lead nitrate induced toxicity in male mice.

Lead is a common environmental occupational toxic metal, known to have indirect oxidative effects. Considering the antioxidant properties of garlic, this study was undertaken to evaluate the therapeutic efficacy of garlic extracts in terms of normalization of altered hematological, biochemical and immunological parameters, and depletion of inorganic lead burden in blood, kidney and brain tissues. Chronic lead nitrate ingestion showed a significant decline in total erythrocyte count, total leukocyte count, hemoglobin concentration, lymphocyte and monocyte content, while neutrophil content increased in lead nitrate treated group. Pb(NO(3))(2) exposure elicited a significant escalation in thiobarbituric acid reactive substances level and depletion in reduced glutathione content and antioxidant enzymes namely, superoxide dismutase and catalase in kidney and brain. Activities of aspartate transaminase, alanine transaminase, acid phosphatase and alkaline phosphatase augmented significantly in kidney and brain of lead exposed mice. Lead nitrate treatment decreased protein content while cholesterol and lead burden increased significantly. A decrease in viability of macrophage, phagocytic index, immunoglobulin level and plaque count were the salient features observed in lead exposed animals. However, oral administration of garlic extracts to Pb(NO(3))(2) treated groups attenuated the deranged parameters to some extent. This indicates that garlic can be a protective regimen for lead toxicity.

Prophylactic efficacy of Coriandrum sativum (Coriander) on testis of lead-exposed mice.

Lead poisoning is a worldwide health problem, and its treatment is under investigation. The aim of this study was to access the efficacy of Coriandrum sativum (coriander) in reducing lead-induced changes in mice testis. Animal exposed to lead nitrate showed significant decrease in testicular SOD, CAT, GSH, total protein, and tissue lead level. This was accompanied by simultaneous increase in the activities of LPO, AST, ALT, ACP, ALP, and cholesterol level. Serum testosterone level and sperm density were suppressed in lead-treated group compared with the control. These influences of lead were prevented by concurrent daily administration of C. sativum extracts to some extent. Treating albino mice with lead-induced various histological changes in the testis and treatment with coriander led to an improvement in the histological testis picture. The results thus led us to conclude that administration of C. sativum significantly protects against lead-induced oxidative stress. Further work need to be done to isolate and purify the active principle involved in the antioxidant activity of this plant.

Amelioration of lead-induced hepatotoxicity by Allium sativum extracts in Swiss albino mice.

Lead is a blue-gray and highly toxic divalent metal that occurs naturally in the earth's crust and is spread throughout the environment by various human activities. The efficacy of garlic (Allium sativum) to reduce hepatotoxicity induced by lead nitrate was evaluated experimentally in male mice. Oral treatment with lead nitrate at a dose of 50 mg/kg body weight daily for 40 days (1/45 of LD50) induced a significant increase in the levels of hepatic aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, acid phosphatase, cholesterol, lipid peroxidation, and lead nitrate. In parallel, hepatic protein levels in lead-exposed mice were significantly depleted. Lead nitrate exposure also produced detrimental effects on the redox status of the liver indicated by a significant decline in the levels of liver antioxidants such as superoxide dismutase, catalase, and glutathione. After exposure to lead nitrate (50 mg/kg body weight for 10 days), the animals received aqueous garlic extract (250 mg/kg body weight and 500 mg/kg body weight) and ethanolic garlic extract (100 mg/kg body weight and 250 mg/kg body weight), and partially restored the deranged parameters significantly. Histological examination of the liver also revealed pathophysiological changes in lead nitrate-exposed group and treatment with garlic improved liver histology. Our data suggest that garlic is a phytoantioxidant that can counteract the deleterious effects of lead nitrate.