학술논문
红景天苷通过抑制NOX2-ROS-MAPKs信号途径保护H2O2诱导的PC12细胞凋亡 / Salidroside protects PC12 cells from H2O2-induced apoptosis via suppressing NOX2-ROS-MAPKs signaling pathway
Document Type
Academic Journal
Author
Source
南方医科大学学报 / Journal of Southern Medical University. 37(2):178-183
Subject
Language
Chinese
ISSN
1673-4254
Abstract
目的 探讨红景天苷保护H2O2诱导PC12细胞凋亡的分子机制.方法 PC12细胞置于含10％马血清、5％胎牛血清的高糖DMEM完全培养基中常规培养.不同浓度的红景天苷预处理细胞2h,然后H2O2作用细胞特定的时间,Western blot检测细胞凋亡相关蛋白PARP,caspase 3的表达及胞内信号分子p38,ERK和JNK的磷酸化水平;DAPI染色检测细胞核的形态改变;ROS检测试剂盒检测胞内ROS的水平;膜质分离实验检测NOX2的两个重要亚基gp91phox和p47phox在胞膜和胞质中的含量;免疫共沉淀实验检测gp91phox和p47phox的结合.结果 红景天苷浓度依赖性的抑制H2O2诱PC12细胞凋亡;减弱H2O2诱导p38,JNK和ERK的磷酸化;减少H2O2诱发的ROS释放;抑制gp91phox和p47phox的结合,进而抑制NOX2的活性.结论 红景天苷通过抑制NOX2-ROS-MAPKs信号通路保护H2O2诱导PC12细胞凋亡.
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.