热休克蛋白70参与肥厚预刺激改善心肌细胞肥大的作用和机制  

The Effect and Mechanism of Heat Shock Protein 70 in Preconditioning Improving Cardiomyocyte Hypertrophy

作　　者：孙军健 俞峰 周巍 王燕华 向科发 张帅 郑毅敏 孙俊波 袁志军 吴峰 SUN Jun-jian;YU Feng;ZHOU Wei;WANG Yan-hua;XIANG Ke-fa;ZHANG Shuai;ZHENG Yi-min;SUN Jun-bo;YUAN Zhi-jun;WU Feng(Department of Orthopedics,The 72nd Group Army Hospital,Huzhou University,Huzhou,Zhejiang,313000,China;Department of Cardiology,The 72nd Group Army Hospital,Huzhou University,Huzhou,Zhejiang,313000,China)

机构地区：[1]湖州学院陆军第七十二集团军医院骨科,浙江湖州313000 [2]湖州学院陆军第七十二集团军医院心内科,浙江湖州313000

出　　处：《现代生物医学进展》2025年第4期622-629,共8页Progress in Modern Biomedicine

基　　金：中华中医药学会项目(2023HH-011);浙江省省医医学会临床医学专项项目(2023ZYC-A35,2023ZYC-A153);湖州市自然科学基金项目(2019YZ11)。

摘　　要：目的:明确热休克蛋白70(Heat shock protein 70,HSP70)在肥厚预刺激改善心肌细胞肥大中的作用和潜在机制.方法:胶原酶消化乳鼠心肌组织,差速贴壁法分离心肌细胞后分组培养.AngⅡ组以1µmol/L血管紧张素Ⅱ(AngⅡ)刺激48 h;Pre-AngⅡ组以12 h的0.5µmol/L的预刺激12 h:常规培养12 h后,再以1µmol/L的AngⅡ刺激48 h.以α-actinin的免疫荧光评价心肌细胞表面积,利用实时荧光定量PCR检测肥厚标志物的基因表达,利用蛋白质免疫印迹检测HSP70及自噬标志蛋白的蛋白表达.结果:相对于AngⅡ组,Pre-AngⅡ组心肌细胞表面积明显减少(P<0.01);ANP、BNP和β-MHC的表达明显降低(P<0.01);HSP70的表达明显增加(P<0.01).HSP70抑制剂VER-155008处理后,肥厚预刺激对心肌细胞肥大的保护作用显著降低(P<0.01);心肌细胞中Beclin 1和LC3Ⅱ/Ⅰ的水平明显减少(P<0.01).结论:肥厚预刺激对心肌细胞肥大的保护作用可能是通过提高HSP70和自噬活性实现的.Objective:To clarify the role and potential mechanism of heat shock protein 70(HSP70)in preconditioning improving cardiomyocyte hypertrophy.Methods:Neonatal rat myocardial tissues were digested with collagenase,and cardiomyocytes were isolated by differential adhesion and then cultured in groups.Cardiomyocytes in the AngⅡgroup was stimulated with 1 pmol/L AngiotensinⅡ(AngⅡ)for 48 hours.Cardiomyocytes in the Pre-AngⅡgroup was preconditioned with 0.5μmol/L AngⅡfor 12 hours,followed by conventional culture for another 12 hours,and then stimulated with 1 pmol/L AngⅡfor 48 hours.The surface area of cardiomyocytes was evaluated byα-actinin immunofluorescence,the gene expression of hypertrophy markers was detected by real-time quantitative PCR,and the protein expression of HSP70 and autophagic markers was detected by western blot.Results:Compared with the AngⅡgroup,the surface area of cardiomyocytes in the Pre-AngⅡgroup was significantly reduced(P<0.01);the expression of ANP,BNP,andβ-MHC was significantly decreased(P<0.01);the expression of HSP70 was significantly increased(P<0.01).After treatment with the HSP70 inhibitor VER-155008,the protective effect of preconditioning on cardiomyocyte hypertrophy was significantly reduced(P<0.01);the levels of Beclin 1 and LC3Ⅱ/Ⅰin cardiomyocytes were significantly decreased(P<0.01).Conclusions:The protective effect of preconditioning on cardiomyocyte hypertrophy may be achieved by increasing the activity of HSP70 and autophagy.

关 键 词：热休克蛋白70 肥厚预刺激 心肌肥厚 心肌细胞 自噬 

分 类 号：R3[医药卫生—基础医学] R364.1