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
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