IN VITRO STUDIES ON THE SYNERGISTIC EFFECT OF MORINDA CITRIFOLIA L. (NONI) AND METHOTREXATE ON CYTOTOXICITY OF HELA CELL LINES

Objective: Cancer remains to be one of the leading causes of death around the world. Modern targeted therapies have undeniably improved cancer patients’ survival, but search still continues for safer and more effective drugs. This work is an attempt to assess the effectiveness of cotreatment of aqueous fruit extract of Morinda citrifolia L. (Noni) on methotrexate (MTX)-induced cytotoxicity on HeLa cancer cell line. Methods: HeLa cells were treated with different concentrations of MTX and Noni alone and in combination for 24 and 48 h and studied for the cytotoxic effects by various approaches. Results: There was a dose-dependent inhibition of growth when treated with MTX in a dose range of 0.045–45.4 μg/ml and Noni for a dose range of 0.3–30 mg/ml for 48 h. The IC50 for MTX was 4.45 μg/ml (1 μM/ml), and for Noni, it was 8 mg/ml while the combination exhibited a more intensive growth inhibitory effect. Our results were confirmed with phase-contrast microscopy, whereby the number of viable cells decreased with an increase in the concentration of Noni. Cells were found to have rounded up and detached from the flask indicating apoptosis. Apoptosis in HeLa cell lines was further confirmed by DNA fragmentation and flow cytometry. Conclusion: It can be concluded that Noni may enhance MTX cytotoxicity by inducing apoptosis and cell cycle arrest. Hence, cotreatment with Noni may reduce the dosage of MTX necessary for inhibiting proliferation of malignant cells and hence decrease the toxic effects of MTX on the system

IMPACT OF ANTIOXIDANT SUPPLEMENTATION ON TOXICITY OF METHOTREXATE-AN IN VITRO STUDY ON ERYTHROCYTES USING VITAMIN E

 Objective: Methotrexate (MTX) is an antimetabolite used widely in cancer and autoimmune diseases as an inhibitor of the enzyme dihydrofolatereductase. Treatment regimens with MTX lead to the formation of reactive oxygen species and are involved in drug-induced toxicity presenting severeside effects. In the present investigation, red blood cells were used as a model to understand drug-mediated hemolysis and alterations in antioxidantdefense system stimulated by MTX. This effort attempts to evaluate, whether pre-supplementation of vitamin E can modulate drug induced toxicity.Methods: Blood samples from healthy subjects were collected and processed to obtain 10% RBC solution in saline. This solution was further treatedwith 80μM MTX in presence and absence of 90μM vitamin E respectively. Treated and control RBC solutions were used to assess percent hemolysis,levels of lipid peroxides, GSH, lactate dehydrogenase and activities of specific antioxidant enzymes.Results: Our study reveals a significant increase in lipid peroxides, reduced glutathione GSH reductase activity after incubation with MTX, lactatedehydrogenase, and a considerable decline in catalase, superoxide dismutase, GSH peroxidase, GSH S-transferase, and GSH reductase after incubationwith MTX.Conclusion: Ameliorative effect of vitamin E supplementation reduces oxidative stress and restores the activities of these antioxidant enzymes,thereby demonstrating the protection rendered by vitamin E. Our data indicates that vitamin E administration during chemotherapy is effective inmodulating the chemotherapy-induced side effects thus stressing on the importance of vitamin E supplementation in combination with chemotherapyduring cancer treatments.Keywords: Antioxidant, Lipid peroxides, Superoxide dismutase, Glutathione, Catalase, Red blood cell

Oxidative stress in sickle cell anemia can be a prognostic marker for disease severity: A case − control study in the western region population of Maharashtra

CONTEXT: Sickle cell anemia (SCA) or sickle cell disease (SCD) is an inherited blood disorder characterized primarily by chronic anemia and periodic episodes of pain. There are reports that increase in oxidative stress may play a significant role in the pathophysiology of SCA. AIM: The present study aims to investigate enzymatic and nonenzymatic antioxidant status in SCA patients to understand the incidence of increased oxidative stress in the populace of tribal Palghar region of Western Maharashtra. SUBJECTS AND METHODS: Patients with SCA (n = 250) and age- and sex-matched healthy persons (n = 250) as controls from the primary health center of Palghar were included in this study. Informed written consent was obtained from all the participants. RESULTS: Activities of enzymatic antioxidants such as glutathione peroxidase, glutathione-S-trasferase, catalase, and level of nonenzymatic antioxidants such as glutathione, Vitamin E and C decreased significantly in SCA participants when compared to controls. The level of lipid peroxides and activity of superoxide dismutase increased significantly above normal in SCA participants. SCA is characterized by the increased generation of reactive-oxygen species resulting in oxidative damage of various cell types, including erythrocytes and this chronically elevated oxidative stress in SCD might play a significant role in the increased autoxidation of Sickle hemoglobin (HbS), increased intravascular hemolysis, ischemia-reperfusion injury, and chronic inflammatory complications. CONCLUSION: The present study indicates that oxidative stress can be considered one of the prognostic markers to evaluate the clinical severity of the SCA participants

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

Association Study between T2DM and CAPN10 SNP-19 (rs3842570) Polymorphism in Navi Mumbai Population

Genetic research has brought a lot of new knowledge in the area of genetic predisposition of type 2 diabetes mellitus (T2DM). It has been proposed that excessive insulin resistance and obesity are also responsible for the higher incidence of type 2 diabetes. Calpain-10 (CAPN10) is a member of a large family of intracellular proteases. The polymorphism at deletion/insertion SNP19 of this gene influences susceptibility to T2DM. The aim of the study was to determine whether calpain-10 (ins/del SNP19) polymorphism contributes significantly to susceptibility to T2DM in population of Navi Mumbai. The study included randomly selected 75 patients of which 33 had T2DM and 42 served as control subjects. Mean waist-to-hip ratio, HDL, LDL, VLDL, cholesterol, and triglyceride showed no difference whereas mean of age, FBS, and body mass index showed significant differences between the control and diabetes subjects. Genotyping of calpain-10 (ins/del SNP19) polymorphism was performed by polymerase chain reaction method. Among 75 participants, for allele-specific SNP19, genotype frequencies of allele1 (2R-32 bp), heterozygous allele (2R-3R 32 bp), and allele 2 (3R-32 bp) were 20 (26.6%), 36 (48%), and 19 (25.3%) observed, respectively. The results from the present study have indicated that CAPN10 (SNP19) shows no significant association with T2DM and more extensive studies on T2DM using candidate gene approach may provide better preventive measures and potential disease diagnostic tools