Use of haematological signatures in conjunction with conventional biomarkers to assess Reno-protective effects of Gedunin in diabetic nephropathy of Streptozotocin induced diabetic rats.

Purpose: The present study was aimed at analysing the reno-protective potential of Gedunin (a limonoid from Neem leaves) through haematological and serological assays and renal histopathology studies in streptozotocin induced diabetic rats. Method: Four weeks old Wister albino rats were chosen for the study. The rats were divided into four groups of 8 rats each. Group 1 (normal control rats); Group 2 (diabetic control rats); Group 3 (Drug control rats: normal rats treated with 1 mg/kg body weight of Gedunin thrice a week through oral gavage for 2 weeks); Group 4 (diabetic rats treated with Gedunin similar to Group 3). Diabetes was induced using a single intraperitoneal injection of streptozotocin (50 mg/kg) in the pre-determined groups. After completion of treatment, the rats were sacrificed and haematological and serological parameters including functional indices of blood cells and renal markers were evaluated through spectrophotometric methods. Renal histology studies were carried out and images were captured on Olympus SC 100, 10 Mega Pixel camera. Results: Streptozotocin induced diabetes lead to marked reduction in the RBC count, haemoglobin, PCV, WBC count, lymphocyte, elevated levels of fasting blood glucose, kidney function markers such as urea, creatinine, uric acid in rats. Administration of Gedunin significantly (P < 0.05) ameliorated the elevated levels of fasting glucose, RDW%, MCHC%, NLR% and platelet count. A significant reversal of renal tissue damage was also observed. Conclusion: The restoration of renal serological markers and reversal of renal tissue injuries are all indicative of promising reno-protective potential of Gedunin. Remarkable improvement of haematological parameters supports their use as reno-protection markers.

A review on interplay between obesity, lipoprotein profile and nutrigenetics with selected candidate marker genes of type 2 diabetes mellitus.

BACKGROUND: Type 2 diabetes mellitus, a rapidly growing epidemic, and its frequently related complications demand global attention. The two factors commonly attributed to the epidemic are genetic factors and environmental factors. Studies indicate that the genetic makeup at an individual level and the environmental aspects influence the occurrence of the disease. However, there is insufficiency in understanding the mechanisms through which the gene mutations and environmental components individually lead to T2DM. Also, discrepancies have often been noted in the association of gene variants and type 2 diabetes when the gene factor is examined as a sole attribute to the disease. STUDY: In this review initially, we have focused on the proposed ways through which CAPN10, FABP2, GLUT2, TCF7L2, and ENPP1 variants lead to T2DM along with the inconsistencies observed in the gene-disease association. The article also emphasizes on obesity, lipoprotein profile, and nutrition as environmental factors and how they lead to T2DM. Finally, the main objective is explored, the environment-gene-disease association i.e. the influence of each environmental factor on the aforementioned specific gene-T2DM relationship to understand if the disease-causing capability of the gene variants is exacerbated by environmental influences. CONCLUSION: We found that environmental factors may influence the gene-disease relationship. Reciprocally, the genetic factors may alter the environment-disease relationship. To precisely conclude that the two factors act synergistically to lead to T2DM, more attention has to be paid to the combined influence of the genetic variants and environmental factors on T2DM occurrence instead of studying the influence of the factors separately.

Functional relevance of Gedunin as a bona fide ligand of NADPH oxidase 5 and ROS scavenger: An in silico and in vitro assessment in a hyperglycemic RBC model.

Clinical evidence suggests that type 2 diabetes therapy can greatly benefit from the suppression of reactive oxygen species generation and the activation or restoration of cellular antioxidant mechanisms. In human, NADPH oxidase (NOX) is the main producer of reactive oxygen species (ROS) that supress the activity of endogenous antioxidant enzymes. In the present study, the antioxidant potential of Gedunin was studied. In silico findings reveal its strong binding affinity with NOX5 C terminal HSP90 binding site that disrupts NOX5 stability and its ability to generate ROS, leading to restoration antioxidant enzymes activities. It was found that Gedunin suppressed hyperglycaemia induced oxidative stress in an in vitro RBC model and markedly reversed glucose induced changes including haemoglobin glycosylation and lipid peroxidation. A significant restoration of activities of cellular antioxidant enzymes; superoxide dismutase, catalase and glutathione peroxidase in the presence of Gedunin revealed its ability to reduce oxidative stress. These results substantiated Gedunin as a bona fide inhibitor of human NOX5 and a ROS scavenging antioxidant with promising therapeutic attributes including its natural origin and inhibition of multiple diabetic targets.

Antioxidants as precision weapons in war against cancer chemotherapy induced toxicity - Exploring the armoury of obscurity.

Cancer is the leading cause of mortality worldwide, accounting for almost 13% of deaths in the world. Among the conventional cancer treatments, chemotherapy is most frequently carried out to treat malignant cancer rather than localised lesions which is amenable to surgery and radiotherapy. However, anticancer drugs are associated with a plethora of side effects. Each drug, within every class, has its own set of adverse reactions which may cause patient incompliance and deterioration of the quality of life. One of the major causes of adverse reactions, especially for drugs targeting DNA, is the excessive production of reactive oxygen species (ROS) and subsequent build up of oxidative stress. To curb these undesired side effects, several dietary supplements have been tested, amongst which antioxidants have gained increasing popularity as adjuvant in chemotherapy. However, many oncologists discourage the use of antioxidant rich food supplements because these may interfere with the modalities which kill cancer by generating free radicals. In the present review, all studies reporting concomitant use of several antioxidants with chemotherapy are indiscriminately included and discussed impartially. The effect of supplementation of thirteen different antioxidants and their analogues as a single agent or in combination with chemotherapy has been compiled in this article. The present review encompasses a total of 174 peer-reviewed original articles from 1967 till date comprising 93 clinical trials with a cumulative number of 18,208 patients, 56 animal studies and 35 in vitro studies. Our comprehensive data suggests that antioxidant has superior potential of ameliorating chemotherapeutic induced toxicity. Antioxidant supplementation during chemotherapy also promises higher therapeutic efficiency and increased survival times in patients.

Exploring the effect of vitamin E in cancer chemotherapy-A biochemical and biophysical insight.

Many oncologists contend that patient undergoing chemotherapy must avoid antioxidant supplementation as it may interfere with the activity of the drug. In the present investigation, we have explored the influence of vitamin E, a well-known antioxidant on Camptothecin (CPT), a potent anti-cancer drug induced cell apoptosis and death of cervical cancer cells. HeLa cells were treated with different concentrations of CPT in presence and absence of 100 µm vitamin E. Treated cells were subjected to cytotoxicity studies, catalase assay, DNA fragmentation assay, clonogenic assay and flow cytometry based apoptosis detection. Also, Raman spectroscopy a label free technique which provides global information, in conjunction with multivariate tools like PCA, PCLDA and FDA, was investigated to explore vitamin E supplementation induced alterations. Our data based on biochemical and biophysical experimental analysis reveals that CPT causes DNA damage along with protein and lipid alteration culminating in cell death. Importantly, Raman spectroscopic analysis could uniquely differentiate the cluster of control and vitamin E control from CPT and CPT + Vit E treated cells. We conclusively prove that presence of vitamin E at 100 µM concentration shows promising antioxidant activity and displays no modulatory role on CPT induced effect, thereby causing no possible hindrance with the efficacy of the drug. Vitamin E may prove beneficial to alleviate chemotherapy associated side effects in patients during clinical settings which may open the doors further for subsequent exploration in in vivo preclinical studies.

An in vitro study of the ameliorative role of α-tocopherol on methotrexate-induced oxidative stress in rat heart mitochondria.

BACKGROUND: Methotrexate (MTX) is a folic acid antagonist widely used as a cytotoxic chemotherapeutic agent for many malignancies. Its use is limited due to wide-ranging toxicities. MTX generates reactive oxygen species. Here, we investigated the efficacy of vitamin E supplementation on MTX-induced alterations in vitro. METHODS: Rat heart mitochondria were isolated and used to assess the extent of swelling, lipid peroxidation and alterations in mitochondrial-specific enzyme activities caused by the addition of 80 µM MTX in the presence and absence of 1.2 µM vitamin E. Control for both groups was maintained. RESULT: MTX substantially affects mitochondrial function by increasing lipid peroxidation and mitochondrial swelling. Significant losses in the activities of the tricarboxylic acid cycle enzymes, complex I, II and IV, and an increase in the activity of calcium ATPase were observed in MTX-treated rat mitochondria. Enrichment of the media with vitamin E led to a reduction in swelling and restoration of enzyme activities. CONCLUSIONS: The present study suggests that vitamin E plays a vital role in suppressing MTX-induced mitochondrial toxicity, and affords protection either by reversing the decline of antioxidants or by direct scavenging of free radicals.

Amelioration of glucose induced hemolysis of human erythrocytes by vitamin E.

Cells under aerobic condition are always threatened with the insult of reactive oxygen species, which are efficiently taken care of by the highly powerful antioxidant systems of the cell. The erythrocytes (RBCs) are constantly exposed to oxygen and oxidative stress but their metabolic activity is capable of reversing the injury under normal conditions. In vitro hemolysis of RBCs induced by 5, 10 and 20mM glucose was used as a model to study the free radical induced damage of biological membranes in hyperglycemic conditions and the protection rendered by vitamin E on the same. RBCs are susceptible to oxidative damage, peroxidation of the membrane lipids, release of hemoglobin (hemolysis) and alteration in activity of antioxidant enzymes catalase and superoxide dismutase. The glucose induced oxidative stress and the protective effect of vitamin E on cellular membrane of human RBCs manifested as inhibition of membrane peroxidation and protein oxidation and restoration of activities of superoxide dismutase and catalase, was investigated. Thiobarbituric acid reactive substances are generated from decomposition of lipid peroxides and their determination gives a reliable estimate of the amount of lipid peroxides present in the membrane. Vitamin E at 18 µg/ml (normal serum level) strongly enhanced the RBC resistance to oxidative lysis leading to only 50-55% hemolysis in 24h, whereas RBCs treated with 10 and 20mM glucose without vitamin E leads to 70-80% hemolysis in 24h. Levels of enzymic antioxidants catalase, superoxide dismutase and nonenzymic antioxidants glutathione showed restoration to normal levels in presence of vitamin E. The study shows that vitamin E can protect the erythrocyte membrane exposed to hyperglycemic conditions and so a superior antioxidant status of a diabetic patient may be helpful in retarding the progressive tissue damage seen in chronic diabetic patients.