Zhi-Lin Qi1, Yin-Hua Liu, Shi-Mei Qi, Lie-Feng Ling, Zun-Yong Feng, Qiang Li. 1. Department of Biochemistry1,Wannan Medical College, Wuhu 241002, China; 3Anhui Provincial Key Laboratory of Active Biological Macro-molecules, Wuhu 241002, China. E-mail: 422627721@qq.com.
Abstract
OBJECTIVE: To investigate the molecular mechanism by which salidroside protects PC12 cells from H2O2-induced apoptosis. METHODS: PC12 cells cultured in DMEM supplemented with 10% horse serum and 5% fetal bovine serum were pretreated with different doses of salidroside for 2 h and then stimulated with H2O2 for different lengths of time. The expression levels of PARP and caspase 3 and the phosphorylation of p38, ERK and JNK were determined with Western blotting. The cell nuclear morphology was observed after DAPI staining. The production of ROS was detected using a ROS detection kit, and the levels of gp91phox and p47phox in the membrane and cytoplasm were detected by membrane-cytoplasm separation experiment; the binding between gp91phox and p47phox was assayed by coimmunoprecipitation experiment. RESULTS: Salidroside dose-dependently suppressed cell apoptosis, lowered phosphorylation levels of p38, ERK and JNK, inhibited the production of ROS, reduced the binding between gp91phox and p47phox, and inhibited the activity of NOX2 in PC12 cells exposed to H2O2. CONCLUSION: Salidroside protects PC12 cells from H2O2-induced apoptosis at least partly by suppressing NOX2-ROS-MAPKs signaling pathway.
OBJECTIVE: To investigate the molecular mechanism by which salidroside protects PC12 cells from H2O2-induced apoptosis. METHODS: PC12 cells cultured in DMEM supplemented with 10% horse serum and 5% fetal bovine serum were pretreated with different doses of salidroside for 2 h and then stimulated with H2O2 for different lengths of time. The expression levels of PARP and caspase 3 and the phosphorylation of p38, ERK and JNK were determined with Western blotting. The cell nuclear morphology was observed after DAPI staining. The production of ROS was detected using a ROS detection kit, and the levels of gp91phox and p47phox in the membrane and cytoplasm were detected by membrane-cytoplasm separation experiment; the binding between gp91phox and p47phox was assayed by coimmunoprecipitation experiment. RESULTS:Salidroside dose-dependently suppressed cell apoptosis, lowered phosphorylation levels of p38, ERK and JNK, inhibited the production of ROS, reduced the binding between gp91phox and p47phox, and inhibited the activity of NOX2 in PC12 cells exposed to H2O2. CONCLUSION:Salidroside protects PC12 cells from H2O2-induced apoptosis at least partly by suppressing NOX2-ROS-MAPKs signaling pathway.