Protective mechanisms of melatonin against hydrogen peroxide induced toxicity in human bone-marrow derived mesenchymal stem cells.
Can J Physiol Pharmacol. 2016 Dec 23. Epub 2016 Dec 23. PMID: 28177678
Many obstacles compromise the efficacy of bone marrow mesenchymal stem cells (MSCs) by inducing apoptosis in the grafted MSCs. Current study investigates the effect of melatonin on important mediators involved in survival of BM-MSCs in hydrogen peroxide (H2O2) apoptosis model.In brief, BM-MSCs were isolated treated with melatonin and then exposed to H2O2.Their viability was assessed by MTT assay and apoptotic fractions were evaluated through Annexin V, Hoechst staining and ADP/ATP ratio. Oxidative stress biomarkers including: ROS, total antioxidant power (TAP), superoxide dismutase (SOD) and catalase (CAT) activity, glutathione (GSH), thiol molecules and lipid peroxidation (LPO) levels were determined. Secretion of inflammatory cytokines (TNF-alpha and IL-6) were measured by ELISA assay.The proteins expression of caspase-3, Bax and Bcl-2, was also evaluated by western blotting.Melatonin pretreatment significantly increased viability and decreased apoptotic fraction of H2O2 exposed BM-MSCs.Melatonin also decreased ROS generation as well as increasing the activity of SOD and CAT enzymes and GSH content. Secretion of inflammatory cytokines in H2O2 exposed cells were also reduced by melatonin. Expression of caspase-3 and Bax proteins in H2O2 exposed cells was diminished by melatonin pretreatment. The findings suggest that melatonin may be an effectively protective agent against H2O2-induced oxidative stress and apoptosis in MSC.