缺氧后适应中HSP90对补体介导的H9c2心肌细胞缺氧/复氧损伤的作用  

Effects of HSP90 on complement-mediated hypoxia/reoxygenation injury of H9c2 cardiomyocytes during hypoxic postconditioning

作　　者：黄政 涂荣会[2] 钟国强[2] 方存明 马小林 胡学俊 HUANG Zheng;TU Rong-hui;ZHONG Guo-qiang;FANG Cun-ming;MA Xiaolin;HU Xue-jun(Department of Cardiovascular Medicine,Xuancheng People's Hospital,Xuancheng 242000,China;Department of Cardiovascular Medicine,First Affiliated Hospital of Guangxi Medical University,Nanning 530022,China)

机构地区：[1]宣城市人民医院心血管内科,安徽宣城242000 [2]广西医科大学第一附属医院心内科,广西南宁530022

出　　处：《中国病理生理杂志》2020年第11期1960-1965,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金地区基金项目(No.81560068);宣城市科技计划项目(No.1817)。

摘　　要：目的:探讨缺氧后适应(HPC)中热休克蛋白90(HSP90)对补体介导的大鼠H9c2心肌细胞缺氧/复氧(H/R)损伤的作用及机制。方法:大鼠H9c2心肌细胞按处理方式不同分为7组:(1)对照组(常氧培养10 h);(2)H/R组(缺氧4 h后复氧6 h);(3)HPC组(缺氧4 h后行复氧5 min/缺氧5 min,重复3次,再复氧6 h);(4)HPC+格尔德霉素(GA)组(缺氧前20 min加入1μmol/L的HSP90特异性抑制剂GA后行HPC);(5)阴性对照组(转染空载质粒后行HPC);(6)C3过表达组(转染C3a质粒后行HPC);(7)C5过表达组(转染C5a质粒后行HPC)。采用Hoechst 33242染色观察细胞形态变化,流式细胞术检测细胞凋亡,Western blot检测HSP90、C3a、C5a、NF-κB p65、TNF-α、IL-1β、IL-6、Bcl-2和Bax蛋白的表达。结果:HPC在上调HSP90的同时显著减少H/R诱导的心肌细胞凋亡,并抑制C3a、C5a、NF-κB p65、TNF-α、IL-1β、IL-6和Bax的表达,增加Bcl-2的表达,上述效应可被GA阻断。过表达C3a和C5a后HPC对NF-κB p65表达及细胞凋亡的抑制作用被抵消。结论:HPC中HSP90通过补体/NF-κB信号通路减轻H9c2心肌细胞H/R损伤。AIM:To investigate the effects and mechanisms of heat shock protein 90(HSP90)on comple⁃ment-mediated hypoxia/reoxygenation(H/R)injury of rat H9c2 cardiomyocytes during hypoxic postconditioning(HPC).METHODS:Rat H9c2 cardiomyocytes were divided into 7 groups according to different treatments:(1)control group(cultured for 10 h under normal oxygen);(2)H/R group(hypoxia for 4 h and reoxygenation for 6 h);(3)HPC group(3 cycles of 5 min H/R after hypoxia for 4 h,followed by reoxygenation for 6 h);(4)HPC+geldanamycin(GA)group(1μmol/L HSP90 inhibitor GA was added 20 min before HPC);(5)negative control group(empty plasmid was transfected before HPC);(6)C3 over-expression group(C3a plasmid was transfected before HPC);(7)C5 over-expression group(C5a plasmid was transfected before HPC).Morphological changes of the H9c2 cells were detected by Hoechst 33242 staining.The effects of HPC on the apoptosis of H9c2 cells were examined by flow cytometry.The protein levels of HSP90,C3a,C5a,NF-κB p65,TNF-α,IL-1β,IL-6,Bcl-2 and Bax were determined by Western blot.RESULTS:With up-regulation of HSP90,HPC significantly reduced H/R-induced apoptosis of the H9c2 cells,inhibited the expres⁃sion of C3a,C5a,NF-κB p65,TNF-α,IL-1β,IL-6 and Bax,and increased the expression of Bcl-2.These effects were blocked by GA.The inhibitory effects of HPC on NF-κB p65 expression and H9c2 cell apoptosis were offset after over-expression of C3a or C5a.CONCLUSION:HSP90 attenuates H/R injury of H9c2 cardiomyocytes by inhibiting comple⁃ment-NF-κB signaling pathway during HPC.

关 键 词：缺氧后适应 缺血再灌注损伤 热休克蛋白90 补体 炎症 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R363.2[医药卫生—基础医学]