白藜芦醇通过ROS-JNK通路抑制晚期糖基化终末产物诱导PC12细胞凋亡

Resveratrol inhibits advanced glycation end products induced PC12 cell apoptosis through ROS-JNK pathway

目的探讨白藜芦醇(resveratrol,RSV)对晚期糖基化终末产物(AGEs)诱导大鼠嗜铬细胞瘤(PC12)细胞损伤的保护作用及可能的分子机制。方法MTS检测不同浓度AGEs(0、50、100、200、400 g·L^(-1))和不同浓度RSV(0、5、10、20、40μmol·L^(-1))对细胞存活率的影响。AGEs(400 g·L^(-1))和RSV(10μmol·L-1)联合处理PC12细胞,TUNEL染色检测细胞凋亡;流式细胞术检测细胞凋亡和活性氧(ROS)水平;Western blot检测相关蛋白(p-JNK、JNK、PUMA、Bcl-2、Bax和Caspase-3)的表达。JNK特异性磷酸化抑制剂(SP600125)预处理,流式细胞术观察细胞凋亡。结果与正常组比较,AGEs组细胞存活率降低,细胞凋亡率和ROS水平增加,p-JNK、PUMA、Bax和caspase-3表达增加,Bcl-2表达降低;与AGEs组比较,AGEs+RSV组细胞存活率升高,细胞凋亡率和ROS水平降低,p-JNK、PUMA、Bax和caspase-3表达降低,Bcl-2表达增加。SP600125可部分逆转AGEs对PC12细胞凋亡的作用。结论RSV可显著抑制AGEs诱导的PC12细胞凋亡,其机制可能与激活ROS-JNK通路有关。

Aim To investigate the effect of resveratrol(RSV)on apoptosis of PC12 cells induced by advanced glycation end products(AGEs)and its possible molecular mechanism.Methods MTS assay was used to detect the effects of AGEs(0,50,100,200,400 g·L^(-1))and RSV(0,5,10,20,40μmol·L^(-1))on cell viability.PC12 cells were treated with AGEs(400 g·L^(-1))and RSV(10μmol·L^(-1)).TUNEL staining was used to detect apoptosis;flow cytometry was used to detect apoptosis and reactive oxygen species(ROS);Western blot was used to detect protein expression of p-JNK,JNK,PUMA,Bcl-2,Bax and Caspase-3.Apoptosis was observed by flow cytometry after pretreatment with JNK specific phosphorylation inhibitor(sp600125).Results Compared with normal control group,the cell viability of AGEs group decreased,the apoptotic rate and ROS levels increased,the expressions of p-JNK,PUMA,Bax and Caspase-3 protein increased,the expression of Bcl-2 protein decreased;Compared with AGEs group,the cell viability of the AGEs+RSV group increased,the levels of apoptotic rate and ROS were reduced,the expressions of p-JNK,PUMA,Bax and caspase-3 protein decreased,the expression of Bcl-2 protein increased.Sp600125 could partially reverse the effect of AGEs on PC 12 cell apoptosis.Conclusions RSV can significantly inhibit the apoptosis of PC 12 cells induced by AGEs,which may be related to the activation of ROS-JNK pathway.