PI3K/Akt通路在槲皮素后处理减轻心肌细胞缺氧/复氧损伤中的作用  

Role of PI3K/Akt signaling pathway in protective effect of quercetin postconditioing alleviating hypoxia/reoxygenation injury of cardiomyocytes

作　　者：王赟[1] 张宗泽[1] 张婧婧[1] 陈莹莹[1] 吴云[1] 王焱林[1] WANG Yun;ZHANG Zongze;ZHANG Jingjing;CHEN Yingying;WU Yun;WANG Yanlin(Zhongnan Hospital of Wuhan University,Wuhan 430071,China)

机构地区：[1]武汉大学中南医院

出　　处：《山东医药》2019年第23期6-9,共4页Shandong Medical Journal

摘　　要：目的探讨槲皮素后处理对大鼠心肌细胞缺氧/复氧(H/R)损伤细胞凋亡的影响及其与磷脂酰肌醇3-激酶/丝氨酸-苏氨酸蛋白激酶(PI3K/Akt)的关系。方法将体外培养的乳鼠心肌细胞随机分为正常对照组(Con组)、H/R组、H/R+槲皮素组(H/R+Que组)、H/R+槲皮素+抑制剂组(H/R+Que+W组)共4组。Con组细胞正常培养6 h;H/R组仅构建H/R模型;H/R+Que组于缺氧结束时,在DMEM培养基中加入终浓度为40μmol/L的槲皮素,随即复氧4 h;H/R+Que+W组于缺氧结束时,在DMEM培养基中加入PI3K抑制剂Wortamannin 100 nmol/L和槲皮素40μmol/L,复氧4 h。采用MTT法测定细胞活力;比色法测定培养液LDH活性;流式细胞仪检测细胞凋亡;Western blotting法检测磷酸化蛋白激酶B(p-Akt)、B细胞淋巴瘤2(Bcl-2)、Bcl-2基因家族中的促凋亡基因(Bax)蛋白表达。结果与Con组比较,H/R组细胞存活率降低,LDH活性和细胞凋亡率升高,p-Akt表达下调,Bcl-2/Bax降低(P均<0.05)。与H/R组比较,H/R+Que组细胞存活率升高,LDH活性和细胞凋亡率降低,p-Akt表达上调,Bcl-2/Bax增高(P均<0.05)。与H/R+Que组比较,H/R+Que+W组细胞存活率降低,LDH活性和细胞凋亡率升高,p-Akt表达下调,Bcl-2/Bax降低(P均<0.05)。结论槲皮素后处理通过激活PI3K/Akt信号途径减轻H/R引起的心肌细胞凋亡。Objective To investigate the effects of quercetin(Que)postconditioing on hypoxia/reoxygenation(H/R)injury-induced apoptosis in primary cultured cardiomyocytes from neonatal rats and its relationship withphosphatidy linositol 3-kinase/Akt PI3K/Akt.Methods Primary cultured cardiomyocytes from neonatal rats were randomly divided into the normal control group(Con group),H/R group,H/R+Que group,and H/R+Que+inhibitor group(H/R+Que+W group).The cells in the Con group were cultured for 6 h;we only established the H/R models in the H/R group;cells in the H/R+Que group were added with Que at a final concentration of 40μmol/L in DMEM medium at the end of hypoxia and followed by reoxygenation for 4 h;in the H/R+Que+W group,the PI3K inhibitor Wortamannin(100 nmol/L)and Que(40μmol/L)were added to the DMEM medium,followed by reoxygenation for 4 h.Cell viability was determined by MTT assay;LDH activity was determined by colorimetric assay;the apoptosis was detected by flow cytometry;the phosphorylated protein kinase B(p-Akt),B-cell lymphoma 2(Bcl-2),and a pro-apoptotic gene(Bax)protein expression in the Bcl-2 gene family were detected by Western blotting.Results Compared with the Con group,the cell survival rate decreased,LDH activity and apoptosis rate increased,p-Akt expression was down-regulated,and Bcl-2/Bax decreased in the H/R group(all P<0.05).Compared with H/R group,the cell survival rate increased,LDH activity and apoptosis rate decreased,p-Akt expression was up-regulated,and Bcl-2/Bax increased in the H/R+Que group(all P<0.05).Compared with the H/R+Que group,the cell survival rate decreased,LDH activity and apoptosis rate increased,p-Akt expression was down-regulated,and Bcl-2/Bax decreased in the H/R+Que+W group(all P<0.05).Conclusion Que postconditioning can attenuate H/R-induced cardiomyocyte apoptosis by activating PI3K/Akt signaling pathway.

关 键 词：槲皮素 缺氧 复氧 心肌细胞 PI3K/AKT信号通路 细胞凋亡 

分 类 号：R541[医药卫生—心血管疾病] R363.2[医药卫生—内科学]