摘要
目的探讨白藜芦醇(RES)对人肝癌细胞HepG2凋亡的作用及其影响机制。方法选取对数生长期人肝癌细胞HepG2,分为对照组(DMSO)和RES组(40μmol/L),通过MTT检测RES对HepG2细胞增殖的影响;应用JC-1染色观察RES对HepG2细胞线粒体膜电位的影响;使用Western blot检测RES对HepG2细胞线粒体自噬相关蛋白PINK1及Parkin的影响;检测HepG2细胞自噬及凋亡相关蛋白Beclin 1及Bcl-2表达情况;使用TUNEL染色观察RES对HepG2细胞的凋亡作用。结果MTT检测结果显示,经RES处理后12及24 h后,HepG2细胞增殖被明显抑制;JC-1染色结果显示,RES处理HepG2细胞12及24 h后,HepG2细胞线粒体膜电位显著降低;Western blot结果显示,RES处理HepG2细胞12及24 h后,线粒体自噬相关蛋白PINK1及Parkin显著上调;RES处理HepG2细胞12及24 h后,Beclin 1蛋白表达显著升高而Bcl-2蛋白表达量明显降低;TUNEL染色结果显示,经RES处理HepG2细胞12及24 h后,HepG2细胞的凋亡被激活。结论RES可通过PINK1/Parkin介导HepG2细胞线粒体自噬,从而调节Beclin 1/Bcl-2复合体的表达,激活HepG2细胞的凋亡。
Objective To investigate the effect of resveratrol(RES)on the apoptosis of human liver cancer cells HepG2 and its mechanism.Methods The human liver cancer cells HepG2 in logarithmic growth phase were selected,and the effect of RES on the proliferation of HepG2 cells was detected by MTT.The effect of RES on the mitochondrial membrane potential of HepG2 cells was observed by JC-1 staining.Western blot was used to detect the expression of mitophagy-related protein PINK1 and Parkin;the expression of autophagy and apoptosis-related proteins Beclin 1 and Bcl-2 in HepG2 cells were detected;TUNEL staining was used to observe the apoptosis effect of RES on HepG2 cells.Results MTT results showed that after RES treatment for 12 and 24 h,HepG2 cell proliferation was significantly inhibited.JC-1 results showed that after 12 and 24 h of RES treatment,the mitochondrial membrane potential of HepG2 cells was significantly reduced.Western blot results showed that HepG2 cells were treated by RES for 12 and 24 h,mitochondrial autophagy-related proteins PINK1 and Parkin were significantly up-regulated.After 12 and 24 h of RES treatment of HepG2 cells,Beclin 1 protein expression was significantly increased and Bcl-2 protein expression was significantly reduced.TUNEL staining results showed that HepG2 cells were activated by RES for 12 and 24 h.Conclusion RES could mediate mitochondrial autophagy of HepG2 cells through PINK1/Parkin,thereby regulating the expression of Beclin 1/Bcl-2 complex and activating HepG2 cell apoptosis.
作者
郑修齐
吕小会
张鑫
赵嫄
熊朝刚
ZHENG Xiu-qi;LüXiao-hui;ZHANG Xin;ZHAO Yuan;XIONG Chao-gang(Xi’an Chest Hospital,Xi'an,Shaanxi 710100,China)
出处
《热带医学杂志》
CAS
2021年第4期452-455,I0002,共5页
Journal of Tropical Medicine
基金
陕西省科技计划自然科学基础研究项目(2017JQ8016)。