Protective Effects Of Resveratrol On Oxidative Stress Injury In U251-PC12 Cell Coculture System

OBJECTIVE In this research,the PC12 cells micro-environment injury model was established to simulate the nerve cells in the secondary injury stage after spinal cord injury（SCI）.To observe and determine the effects of resveratrol（RSV）pretreatment at different concentrations on damaged cell models,and investigate the possible mechanism of its neuroprotective effect.METHODS In this study,the U251 cells were induced to oxidative stress by 400μM H₂O₂,then the culture fluid was used to stimulate PC12 cells induced by H₂O₂to establish secondary injury of PC12 cells under oxidative stress in vitro.Meanwhile,low,medium and high concentrations（5μM,10μM and 50μM）of RSV treated the secondary damaged PC12 cells.The CCK-8 method was used to test the cell viability.The ROS kit was used to detect the distribution of ROS and the secretion content.The degree of early apoptosis was explored by detecting the changes of mitochondrial membrane potential.The nucleus were stained with Hoechst 33342 to detect the apoptosis rate of each group.The expressions of MAPK inflammation signaling pathway related proteins,were detected by Western Blot,including MAPK p38,IL-6,IL-1β,and TNF-α.Besides,the expressions of Bcl-2 and Bax apoptosis-related proteins were detected to explore the specific signal transduction mechanism of RSV in playing a neuroprotective role.RESULTS RSV significantly increased the viability of PC12 cells damaged by micro-environment,decreased the content and distribution of ROS in the damaged cells,increased the mitochondrial membrane potential of the damaged cells,and reduced the apoptosis rate.RSV significantly inhibited the expression of MAPK inflammation signaling pathway related proteins and apoptosis related proteins at the transcription level,thereby exerting its neuroprotective function.CONCLUSION RSV exerts its neuroprotective function by improving the vitality of nerve cells and inhibiting oxidative stress,neuroinflammation and apoptosis after cell injury,which provides evidence for clinical use of RSV to treat nerve cell injury after SCI.