摘要
为探讨缺氧后适应(hypoxic postconditioning,HPC)中热休克蛋白70(heat shock protein 70,HSP70)对心肌细胞缺氧/复氧(hypoxia/reoxygenation,H/R)损伤的作用及其机制,将H9C2心肌细胞分为4组:对照组(常氧培养10 h)、H/R组(缺氧4 h后复氧6 h)、HPC组(缺氧4 h后行复氧5 min/缺氧5 min,共3次,再复氧6 h)、HPC+槲皮素(quercetin,Quer)组(缺氧前1 h加50μmol/L的HSP70抑制剂Quer后行HPC)。采用Hoechst 33242染色观察细胞形态变化,FACS检测细胞凋亡情况,Western blotting检测HSP70、TLR4、NF-κB p65、TNF-α、IL-1β、IL-6、B淋巴细胞瘤2基因(B-cell lymphoma 2,Bcl-2)、Bcl-2相关蛋白(Bcl-2-associated X protein,Bax)、Caspase-3蛋白表达水平。结果显示,HPC组HSP70、Bcl-2表达水平较H/R组显著升高(P<0.05),TLR4、NF-κB p65、TNF-α、IL-1β、IL-6、Bax、Caspase-3表达水平较H/R组显著下降(P<0.05);HPC+Quer组HSP70、Bcl-2表达水平较HPC组显著下降(P<0.05),TLR4、NF-κB p65、TNF-α、IL-1β、IL-6、Bax、Caspase-3表达水平较HPC组显著升高(P<0.05)。该研究提示,HPC可通过上调HSP70表达抑制TLR4/NF-κB信号通路,减轻H/R诱导的心肌细胞免疫炎症损伤。
To investigate the effects and mechanisms of heat shock protein 70(HSP70)on the hypoxia/reoxygenation(H/R)injury of cardiomyocytes during hypoxic postconditioning(HPC),H9 C2 cardiomyocytes were divided into four groups:the control group(cultured for 10 hunder normal oxygen),the H/R group(hypoxia for 4 hand reoxygenation for 6 h),the HPC group(3 cycles of 5 min hypoxia/reoxygenation after hypoxia for 4 h,then reoxygenation for 6 h),the HPC+quercetin(Quer)group(50μmol/L HSP70 inhibitor Quer was added 1 hbefore HPC).Morphological changes of cardiomyocytes were examined by Hoechst 33242 staining,and cardiomyocytes apoptosis were examined by FACS.The effects of HPC on the expression of HSP70,TLR4,NF-κB p65,TNF-α,IL-1β,IL-6,B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax)and Caspase-3 were determined by Western blotting.The results showed that the levels of HSP70 and Bcl-2 in the HPC group were significantly higher than those in the H/R group(P<0.05),and TLR4,NF-κB p65,TNF-α,IL-1β,IL-6,Bax,Caspase-3 levels were significantly lower than those in the H/R group(P<0.05).Whereas the levels of HSP70 and Bcl-2 in the HPC+Quer group were significantly lower than those in the HPC group(P<0.05),and TLR4,NF-κB p65,TNF-α,IL-1β,IL-6,Bax,Caspase-3 levels were significantly higher than those in the HPC group(P<0.05).Thus,HPC inhibits the TLR4/NF-κB signaling pathway and alleviates the H/R-induced inflammatory injury of cardiomyocytes by upregulating HSP70 expression.
作者
黄政
涂荣会
钟国强
方存明
马小林
胡学俊
HUANG Zheng;TU Rong-hui;ZHONG Guo-qiang;FANG Cun-ming;MA Xiao-lin;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)
出处
《现代免疫学》
CAS
北大核心
2021年第5期386-391,共6页
Current Immunology
基金
国家自然科学基金地区基金(81560068)
宣城市科技计划项目(1817)。
