A cross talk study on sitagliptin mediated reclamation on TGF ß signalling, DPP 4, miR-29a and miR-24 expression in PCOS rats fed with high fat-high fructose diet.

Polycystic Ovary Syndrome (PCOS) is a multifactorial reproductive, endocrine and metabolic disturbance which is very commonly observed in females of reproductive age group. The disease is still incurable however the use of synthetic drugs in combination with lifestyle is recommended. Accordingly, the present study was conducted to investigate the possible beneficial effects of sitagliptin on PCOS induced rats on control diet (CD)/high fat- high fructose diet (HFFD). PCOS was induced by giving testosterone propionate (TP) for 28 days to both the CD/HFFD rats and treated with STG i.p. for last 15 days. At the end of the experiment lipid profile, inflammatory markers, expression of NF-κB-p65, miR-24 and miR-29a, fibrotic and apoptotic proteins from ovary tissue were examined. Moreover, lipid accumulation and fibrosis of ovary tissue was further confirmed using Sudan III and Masson's trichrome stain. STG treated rats exerted a significant decrease in levels of cholesterol, TG, LDL-C, VLDL-C, IL-6 and TNF-α and increased HDL-C level, miR-24 and miR-29a expression. STG treated groups expressed significantly decreased expression of NF-κB-p65, TGF-ß1, p-Smad 2 and p-Smad 3 followed by no significant changes in the expression of BAX, caspase-9, caspase-3 and Bcl-2 in all the PCOS induced groups. Among all the CD/ HFFD fed groups, rats on HFFD showed more devastating effect which suggests that diet plays a major role in genesis of PCOS. In conclusion, current results reflect the potential impact of STG against dyslipidaemia, inflammation and fibrosis in PCOS rats via regulating dyslipidaemia and fibrosis via DPP 4 mediated miR-29a expression.

Anticancer Activity of Rhizophora mucronata Leaves Extract on Sprague-Dawley Rats: In Vivo Model.

Medicinal plants are now used to treat cancer due to the presence of bioactive compounds. Apart from the plants, mangroves also possess rich bioactive compounds with high medicinal activity. Based on the ethnobotanical attributes of Rhizophora mucronata, we are keen to work with its anticancer activity. The aim of the study is to assess the anticancer activity of methanolic extract of Rhizophora mucronata leaves against breast cancer. Its safety profile for anticancer investigations was therefore confirmed through an acute toxicity assessment. In accordance with OECD guiding principles, the study was approved to evaluate the toxicity, including acute and subacute effects and anticancer activities of methanolic extract of Rhizophora mucronata leaves on Sprague-Dawley rats. In acute toxicity trials, the dose of 1000 mg/kg body weight was determined to be safe and nontoxic even at high dose levels and did not result in any indicators of toxicity or death in the tested groups compared to controls for 14 days. In contrast, rats in a subacute toxicity study were given consistent doses of 100 mg/kg, 200 mg/kg, and 300 mg/kg for a total of 28 days along with a control group. Haematological, biochemical, and histological tests conducted in advance revealed that methanolic extract of Rhizophora mucronata leaves (MERML) at repeated doses of 200 mg/kg and 300 mg/kg was normal and had no significant effects on the treated groups. Rhizophora mucronata extract (250 mg/kg) was successfully used in in vivo trials to stop the growth of breast cancer cells in groups that had been given DMBA. Based on these findings, it has been concluded that methanolic extract of Rhizophora mucronata leaves (MERML) was safe at both higher and lower dosages and could be assessed for pharmacological study.

Bursatella leachii Purple Ink Secretion Concentrate Exerts Cytotoxic Properties against Human Hepatocarcinoma Cell Line (HepG2): In Vitro and In Silico Studies.

Liver cancer is a leading cause of cancer death globally. Marine mollusc-derived drugs have gained attention as potential natural-based anti-cancer agents to overcome the side effects caused by conventional chemotherapeutic drugs during cancer therapy. Using liquid chromatography-mass spectrometry, the main biomolecules in the purple ink secretion released by the sea hare, named Bursatella leachii (B. leachii), were identified as hectochlorin, malyngamide X, malyngolide S, bursatellin and lyngbyatoxin A. The cytotoxic effects of B. leachii ink concentrate against human hepatocarcinoma (HepG2) cells were determined to be dose- and time-dependent, and further exploration of the underlying mechanisms causing the programmed cell death (apoptosis) were performed. The expression of cleaved-caspase-8 and cleaved-caspase-3, key cysteine-aspartic proteases involved in the initiation and completion of the apoptosis process, appeared after HepG2 cell exposure to the B. leachii ink concentrate. The gene expression levels of pro-apoptotic BAX, TP53 and Cyclin D1 were increased after treatment with the B. leachii ink concentrate. Applying in silico approaches, the high scores predicted that bioactivities for the five compounds were protease and kinase inhibitors. The ADME and cytochrome profiles for the compounds were also predicted. Altogether, the B. leachii ink concentrate has high pro-apoptotic potentials, suggesting it as a promising safe natural product-based drug for the treatment of liver cancer.

In vitro cytotoxicity assay of Fucoidan extracted from Turbinaria conoides against cancer cell lines MCF7, A549, and normal cell line L929

Abstract The main aim of the study is to quantify the cytotoxic property of the Fucoidan extracted from the Turbinaria conoides using the MTT assay with the standard fucose. Fucoidan was extracted using the soaked water method and it was determined using the HPLC procedure the obtained Test sample Fucoidan extracted from the Turbinaria conoides and standard fucose was subjected to the cytotoxicity assay against the MCF7 Human breast cancer cell line, A549 lung cancer cell line, and L929 normal mouse fibroblast cell line. From the results it was found that the Test sample showed good IC50 value for MCF7 cell line then A549 with an increasing concentration 24 hours incubation at 37°C The IC50 for MCF7 was 115.21 µg/ml and A549 396.46µg/ml and the Fucoidan extract was checked for its cytotoxicity against the normal mouse fibroblast cell line L929, Fucoidan was found non-lethal to the L929 mouse fibroblast normal cell line. Standard fucose also gave a significant result towards MCF7 and against the L929. This indicates that the Fucoidan extracted from Tubinaria conoides shows better anticancer potential in it. Hence its application can be further extended in the pharmacological fields.

Evaluation of Serum miRNA-24, miRNA-29a and miRNA-502-3p Expression in PCOS Subjects: Correlation with Biochemical Parameters Related to PCOS and Insulin Resistance.

MicroRNAs (miRNAs) are small, endogenous, non-coding, single stranded RNAs which play a role in the regulation of gene expression and function. Therefore, the analysis of differentially expressed miRNAs are of great importance in disease diagnosis. This study is focussed on the differential expression of miRNAs in serum of PCOS subjects compared to control and their correlation with metabolic and endocrine parameters. Anthropometry, hormone concentrations and biochemical characteristics were measured in healthy (n = 20) and PCOS (n = 20) subjects. MiR-24, miR-29a and miR-502-3p were determined in serum by quantitative RT-PCR. The levels of miR-24 was significantly decreased in PCOS subjects (P = 0.00) compared to control. No significant difference was observed in the levels of miR-29a and miR-502-3p in PCOS and control subjects. MiR-24 showed significant inverse correlation with BMI, glucose, insulin, FIRI, HOMA, LH, testosterone, TG, and LH:FSH ratio whereas HDL levels showed significant positive association with miR-24 and miR-29a. LH showed significant negative association with miR-29a. No correlation was observed between the expression of miR-502-3p with any of the studied parameters. The receiver operating characteristic curve for miR-24 alone showed a significant discriminative capacity. The study suggests that serum miR-24 analysis in PCOS patients could be of diagnostic value that can be used as a biomarker for PCOS.

Propugnating Effect of Bark of Rhizophora mucronata Against Different Toxicants Viz Carbon Tetrachloride, Ethanol and Paracetamol on HepG2 Cell Lines.

OBJECTIVE: The aim of the study was to evaluate the hepatoprotective activity of the bark extract (Ethanol: Water) in the ratio of (3:1) of Rhizophora mucronata (BERM) by intoxicating the HepG2 cell lines with different toxicants viz, CCL4, Ethanol and Paracetamol with different concentrations of the extract were used. The HepG2 cell lines were subjected to MTT Assay for studying the cytotoxicity. METHODS: HepG2 cells were plated using 96 well plate in 10% bovine serum, exposed to different toxicants viz, 2% CCl4, 60% Ethanol and 14 mM Paracetamol respectively. The various test concentrations (18.85, 37.5, 75, 150 and 300 µg/ml) of bark extract of Rhizophora mucronata was added and incubated for 24 hours. Medium was removed after incubation period and 0.5 mg/ml MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) was added and again incubated for 4 hours at 37oC. Then MTT was removed the crystals was dissolved in DMSO and absorbance was measured at 570 nm. RESULTS: The result showed that dose dependent increase in percentage of viability at the doses of 18.85, 37.5, 75, 150, 300 µg/ml. The results for the CCl4 intoxicated, at 300 µg/ml of the concentration of the extract, the % of viable cells was found out to be 99.6%, for Ethanol intoxicated, 97.67%, and Paracetamol induced, 75.37%, IC50 was 21.53 µg/ml, 12.61 µg/ml and 21.42 µg/ml respectively. CONCLUSION: Thus, we conclude that, the extract possesses defensive effect against different toxicants and can be used as an alternate drug for hepatotoxicity.

Polyphenols activate energy sensing network in insulin resistant models.

Unhealthy diet deficient in fruits and vegetables but rich in calories is considered to be one factor responsible for the increased prevalence of insulin resistance and type 2 diabetes (T2D). The consumption of fast foods and soft drinks increases fructose consumption per se and this is of major concern since prolonged fructose intake induces insulin resistance and thereby T2D. The energy homeostasis is regulated by a network consisting of "fuel gauze" called AMP-activated protein kinase (AMPK), the NAD+ dependent type III deacetylase (SIRT1) and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) which is disrupted in T2D. The present study was aimed to investigate the action of naringenin and quercetin on energy sensing molecules in insulin resistant models. L6 myotubes and albino Wistar rats were rendered insulin resistant with palmitate and fructose respectively. Naringenin, quercetin or metformin were used for treatment. Fructose and palmitate treatment resulted in insulin resistance as evidenced by decreased glucose transporter 4 (GLUT4) translocation. The translocation of GLUT4, phosphorylation of AMPK and the expression of SIRT1 and PGC-1α which were reduced in insulin resistant cells, were increased upon treatment with polyphenols. Further, naringenin and quercetin showed binding affinity with energy sensing molecules. We conclude that drugs from natural resources that target energy sensing molecules might be helpful to prevent insulin resistance.

Apigenin attenuates hippocampal oxidative events, inflammation and pathological alterations in rats fed high fat, fructose diet.

High calorie diet promotes oxidative stress and chronic low grade inflammation that predispose to brain dysfunction and neurodegeneration. Hippocampus region of the brain has been shown to be particularly sensitive to high calorie diet. We hypothesize that apigenin (API), a flavonoid could attenuate hippocampal derangements induced by high fat-high fructose diet (HFFD). In this study, we investigated the effects of API on oxidative stress and inflammation in the hippocampus, and compared with those of sitagliptin (STG), a standard drug with neuroprotective properties. The markers of oxidative stress and inflammation were examined using biochemical assays, western blotting and immunohistochemistry techniques. HFFD-fed rats showed severe pathological alterations and API treatment rescued the hippocampus from the derangements. API significantly improved the antioxidant machinery, reduced ROS levels and prevented the activation of the stress kinases, inhibitor of kappa B kinase beta (IKKß) and c-Jun NH2 terminal kinase (JNK), and the nuclear translocation and activation of nuclear factor kappa B (NF-κB). The plasma levels of inflammatory cytokines were also reduced. Our findings suggest that hippocampal derangements triggered by HFFD feeding were effectively curtailed by API. Suppression of oxidative stress, NF-κB activation and JNK phosphorylation in the hippocampus are the mechanisms by which API offers neuroprotection in this model.

Troxerutin attenuates diet-induced oxidative stress, impairment of mitochondrial biogenesis and respiratory chain complexes in mice heart.

Mitochondrial abnormality is thought to play a key role in cardiac disease originating from the metabolic syndrome (MS). We evaluated the effect of troxerutin (TX), a semi-synthetic derivative of the natural bioflavanoid rutin, on the respiratory chain complex activity, oxidative stress, mitochondrial biogenesis and dynamics in heart of high fat, high fructose diet (HFFD) -induced mouse model of MS. Adult male Mus musculus mice of body weight 25-30 g were fed either control diet or HFFD for 60 days. Mice from each dietary regimen were divided into two groups on the 16th day and were treated or untreated with TX (150 mg/kg body weight [bw], per oral) for the next 45 days. At the end of experimental period, respiratory chain complex activity, uncoupling proteins (UCP)-2 and -3, mtDNA content, mitochondrial biogenesis and dynamics, oxidative stress markers and reactive oxygen species (ROS) generation were analyzed. Reduced mtDNA abundance with alterations in the expression of genes related to mitochondrial biogenesis and fission and fusion processes were observed in HFFD-fed mice. Disorganized and smaller mitochondria, reduction in complexes I, III and IV activities (by about 55%) and protein levels of UCP-2 (52%) and UCP-3 (46%) were noted in these mice. TX administration suppressed oxidative stress, improved the oxidative capacity and biogenesis and restored fission/fusion imbalance in the cardiac mitochondria of HFFD-fed mice. TX protects the myocardium by modulating the putative molecules of mitochondrial biogenesis and dynamics and by its anti-oxidant function in a mouse model of MS.

Effects of linalool on inflammation, matrix accumulation and podocyte loss in kidney of streptozotocin-induced diabetic rats.

CONTEXT: This study aimed to evaluate the renoprotective action of linalool (LIN) on streptozotocin (STZ)-induced diabetic rats. OBJECTIVE: The pathological changes in diabetic nephropathy (DN) include oxidative stress, renal injury, matrix accumulation and podocyte abnormalities. We investigated the renoprotective actions of LIN, a monoterpene alcohol, present in herbal essential oils in STZ-induced diabetic rats with renal injury. MATERIALS AND METHODS: STZ-diabetic rats were administered LIN (25 mg/kg) for 45 days, after which the activities of key enzymes of glucose metabolism, collagen content, oxidative damage and expression of glucose transporter-1 (GLUT-1), transforming growth factor-ß1 (TGF-ß1), nuclear factor-κBp65 (NF-κB p65) and nephrin were analyzed. RESULTS: Diabetic rats displayed altered glucose metabolism, collagen accumulation and increased TGF-ß1 and NF-κB expression in kidney. LIN treatment restored glucose-metabolizing enzymes, collagen content and GLUT-1 expression and also prevented nephrin loss. LIN also rescued kidney from oxidative stress and inflammation by decreasing the expression of TGF-ß1 and NF-κB. Ultrastructural changes such as basement membrane thickening, reduction in podocyte number and loss of filtration barrier integrity in diabetic rats were mitigated by LIN. DISCUSSION AND CONCLUSION: The results of this study suggest that LIN can attenuate nephropathic changes induced in kidney of diabetic rats. These findings highlight the utility of LIN as a potential drug to treat renal damage in diabetic subjects.

Cissus quadrangularis stem alleviates insulin resistance, oxidative injury and fatty liver disease in rats fed high fat plus fructose diet.

The study evaluated the protective effects of Cissus quadrangularis stem extract (CQEt) on oxidant-antioxidant balance and insulin resistance (IR) in rats fed high fat-high fructose diet (HFFD) and also tested its free-radical scavenging property in vitro. Rats were fed either control diet or HFFD for 15 days, following which the diet was fortified with CQEt at a dose of 10 g/100g diet. After 60 days, HFFD caused deleterious metabolic effects, including hyperglycemia, IR and liver dysfunction. Rats fed HFFD alone showed increased activities of hepatocellular enzymes in plasma, lipid deposition, significant decline in antioxidants, and elevated lipid peroxidation indices and protein carbonyl in liver. CQEt addition significantly improved insulin sensitivity, reduced liver damage and oxidative changes, and brought back the antioxidants and lipids towards normal. Histopathology of the liver confirmed the changes induced by HFFD and the heptoprotective effect of CQEt. The effects of CQEtin vivo were comparable with that of standard drug, metformin. Through in vitro assays, CQEt was found to contain large quantities of polyphenols, vitamins C and E. CQEt exhibited radical scavenging ability in a dose-dependent manner. These data suggest that CQEt affords hepatoprotection by its antioxidant and insulin-sensitizing activities.

Genistein improves liver function and attenuates non-alcoholic fatty liver disease in a rat model of insulin resistance.

BACKGROUND: The high fructose-fed rat is widely used as a model of insulin resistance. Genistein, a soy isoflavone, has been shown to improve insulin sensitivity in this model. The present study investigated whether genistein could prevent fatty liver disease in this model. METHODS: Male Wistar rats were fed a diet containing starch (control) or 60% fructose (insulin-resistant model). Fifteen days later, rats in each dietary group were divided into two groups and were treated with either genistein (1 mg/kg per day) in dimethylsulfoxide (DMSO) or 30% DMSO alone. After 60 days, markers of liver injury, oxidative stress, interleukin (IL)-6, tumor necrosis factor (TNF)-α, lipids, lipoprotein profile, nitrite, and nitrosothiol in the plasma and liver were quantified. Liver sections were examined for 3-nitrotyrosine (3-NT) expression and pathological lesions. RESULTS: Fructose-fed rats displayed hyperlipidemia, significant changes in plasma lipoprotein profile, and increases in IL-6 and TNF-α levels compared with control. In addition, the accumulation of lipids, liver injury, a decline in liver function, inactivation of the glyoxalase system, depletion of antioxidants, and increased 3-NT expression were observed in the fructose-fed group. Administration of genistein to fructose-fed rats significantly reduced these biochemical and histological abnormalities. CONCLUSIONS: Genistein activates the antioxidant profile, decreases IL-6 and TNF-α concentrations, prevents oxidative damage, and ameliorates fatty liver in insulin-resistant rats.