文章摘要

整合素β3在异丙肾上腺素诱导的心肌细胞损伤与凋亡中的作用

作者: 1张海波, 1李运丽, 1艾景雪, 1王亚丹, 2王芳芳
1 河南大学第一附属医院心血管内科,河南 开封 475001
2 中国人民解放军空军军医大学第二附属医院心血管内科,西安 710068
通讯: 王芳芳 Email: wangfftdxn@163.com
DOI: 10.3978/j.issn.2095-6959.2020.08.001
基金: 开封市科技发展计划(1903048)。

摘要

目的:探讨整合素β3(integrin β3,ITGB3)在β-肾上腺素受体(β-adrenergic receptor,β-AR)激动剂异丙肾上腺素(isoprenaline,ISO)诱导的心肌细胞损伤与凋亡中的作用及其机制。方法:心肌细胞随机分为对照组(Con组)、siRNA-ITGB3处理组(Con+si-ITGB3组)、ISO组、ISO+siRNA-ITGB3处理组(ISO+si-ITGB3组)。采用CCK-8检测细胞活性,TUNEL染色检测细胞凋亡,GFP-LC3腺病毒检测细胞自噬流强度,RT-PCR检测ITGB3 mRNA的表达;蛋白质印迹法检测细胞蛋白质水平。结果:在10 μmol/L的ISO刺激后,心肌细胞活力下降、且随刺激时间的延长,细胞活力下降程度增加。siRNA-ITGB3干扰能够下调ISO诱导的ITGB3表达,抑制ISO所致的细胞活力下降。siRNA-ITGB3干扰能够抑制ISO诱导细胞凋亡,抑制促凋亡蛋白cleaved-caspase-3和Bax蛋白表达,上调抗凋亡蛋白Bcl-2表达;siRNA-ITGB3干扰能够增强GFP-LC3表达、促进自噬蛋白Beclin1,LC3-I/LC3-II积累,降低p62水平,抑制ISO所致的细胞自噬水平的下降。结论:ITGB3表达下调能激活细胞自噬,抑制ISO诱导的心肌细胞损伤与凋亡。
关键词: 整合素β3;β-肾上腺素受体;自噬;凋亡

Role of integrin β3 in isoproterenol induced injury and apoptosis in cardiomyocytes

Authors: 1ZHANG Haibo, 1LI Yunl, 1AI Jingxue, 1WANG Yadan, 2WANG Fangfang
1 Department of Cardiology, First Affiliated Hospital of Henan University, Kaifeng Henan 475001, China
2 Department of Cardiology, Second Affiliated Hospital of Air Forced Military Medical University, Xi’an 710068, China

CorrespondingAuthor: WANG Fangfang Email: wangfftdxn@163.com

DOI: 10.3978/j.issn.2095-6959.2020.08.001

Foundation: This work was supported by the Science and Technology Development Project of Kaifeng, China (1903048).

Abstract

Objective: To investigate the role and mechanism of integrin beta 3 (ITGB3) in β‐agonist, adrenoceptor agonist ISO-induced cardiomyocyte injury and apoptosis. Methods: Cardiac myocytes were randomly divided into a control group (Con), a Con+si-ITGB3 group, an isoprinosine (ISO) group, and an ISO+si-ITGB3 group. CCK-8 assay was used to detect cell viability. TUNEL staining was used to detect cell apoptosis. GFP-LC3 adenovirus were used to detect the level of cell autophagy. RT-qPCR was used to detect the mRNA expression of ITGB3. Western blot was used to detect the protein level. Results: After stimulation with 10 μmol/L ISO, the cell viability of cardiomyocytes was reduced gradually with the prolongation of time. After pretreatment with siRNA-ITGB3, siRNA-ITGB3 treatment inhibited the decline of cell viability induced by ISO. Additional, siRNA-ITGB3 treatment inhibited the cell apoptosis induced by ISO, as evidenced by decreased the expression of cleaved caspase 3 and Bax, and increased the expression of Bcl-2. Consistently, siRNA-ITGB3 treatment decreased p62, increased the number of GFP-LC3 positive dots as well as LC3-II/LC3-I ratio and Beclin1 expression. Conclusion: Down-regulation of ITGB3 partially attenuates isoproterenol induced injury and apoptosis via activation of autophagy in cardiomyocytes.
Keywords: integrin β3; beta-adrenergic receptor; autophagy; apoptosis