Effects of Reactive Oxygen Species on Differentiation of Bone Marrow Mesenchymal Stem Cells.

BACKGROUND The low differentiation rates for transplanted stem cells are challenging problems in spinal cord injury (SCI) treatment. Studies have demonstrated that the inhibition of the Notch1 pathway in bone marrow mesenchymal stem cells (BMSCs) contributed to the differentiation of these cells. Research findings that certain antioxidants induce BMSCs to differentiate into neuronal cells suggest that BMSC differentiation is related to the level of reactive oxygen species (ROS) in cells. This study aimed to define the effect of ROS on the differentiation of BMSCs. MATERIAL AND METHODS In this study, after BMSCs were induced with the antioxidant β-mercaptoethanol (β-ME), related proteins were analyzed by Western blotting and immunofluorescence. In order to find the role of ROS in the differentiation, DCFH-DA was used to detect ROS levels in antioxidant-treated BMSCs, H2O2-treated BMSCs, and normal BMSCs, and the expression levels of Notch1 and its downstream transcriptional suppressor Hes1 were analyzed. RESULTS Induced with β-ME, Western blotting and immunofluorescence revealed gradual increases in the expression of Nestin (a neural stem cell-specific protein) and neuron-specific enolase (NSE) but decreases in Notch1 expression. The expression levels of Notch1 and Hes1 were positively correlated with changes in ROS level. CONCLUSIONS These data suggest that the antioxidant-induced differentiation of BMSCs into neurons may be related to ROS-based regulation of the Notch1 signalling pathway.