天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (11): 1133-1137.doi: 10.11958/20211124

• 细胞与分子生物学 • 上一篇    下一篇

SIRT3-FOXO1介导自噬在H9C2心肌细胞缺血再灌注损伤中的保护作用

王尚徐，林先和   

  1. 安徽医科大学第一附属医院心血管内科（邮编230032）
  • 收稿日期:2021-05-12 修回日期:2021-07-07 出版日期:2021-11-15 发布日期:2021-11-19
  • 基金资助:
    安徽省自然科学基金资助项目（2008085MH239）

The protective effect of SIRT3-FOXO1-mediated autophagy on ischemia-reperfusion injury of H9C2 cells

WANG Shang-xu, LIN Xian-he   

  1. Department of Cardiovascular Medicine, the First Affiliated Hospital of Anhui Medical University, Hefei 230032, China
  • Received:2021-05-12 Revised:2021-07-07 Published:2021-11-15 Online:2021-11-19

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摘要： 目的　探究沉默信息调节因子2相关酶类3（SIRT3）-叉头盒转录因子O1（FOXO1）通路介导自噬在心肌细胞缺血再灌注损伤中的作用。方法　慢病毒感染H9C2心肌细胞构建SIRT3过表达稳定株。实验分4组：正常对照（NC）组、缺血再灌注（IR）组、SIRT3过表达+缺血再灌注（S+IR）组、SIRT3过表达+IR+SIRT3抑制剂（S+IR+3-TYP）组。Western blot检测SIRT3、乙酰化FOXO1（Ac-FOXO1）、自噬标记蛋白LC3B、Beclin1及凋亡相关蛋白Bcl-2、Bax的表达；流式细胞分析仪检测细胞凋亡率；全自动生化仪检测细胞培养上清液中乳酸脱氢酶（LDH）水平。结果　与NC组相比，IR组SIRT3表达降低，Ac-FOXO1表达升高，自噬标记蛋白LC3B、Beclin1表达升高，抗凋亡蛋白Bcl-2表达降低，促凋亡蛋白Bax表达升高，LDH水平和细胞凋亡率升高（P＜0.05）。与IR组相比较，S+IR组SIRT3表达升高，Ac-FOXO1表达降低，LC3B、Beclin1表达升高，Bcl-2表达升高，Bax表达降低，LDH水平和细胞凋亡率降低（P＜0.05）；加入SIRT3抑制剂3-TYP后，与S+IR组相比，SIRT3表达水平无明显变化，但Ac-FOXO1表达升高，LC3B、Beclin1表达降低，Bcl-2表达降低，Bax表达升高，LDH水平和细胞凋亡率升高（P＜0.05）。结论　SIRT3-FOXO1通路激活可提高H9C2心肌细胞自噬水平，对心肌缺血再灌注损伤有保护作用。

关键词: 肌细胞, 心脏, 再灌注损伤, 自噬, 细胞凋亡, 叉头框蛋白O1, 沉默信息调节因子2相关酶类3, H9C2细胞

Abstract: Objective　To investigate the role of silent mating type information regulator 2 homolog 3 (SIRT3) -forkhead box O1(FOXO1) pathway-mediated autophagy in myocardial ischemia-reperfusion (IR) injury. Methods　SIRT3 overexpression stable strain was constructed by lentivirus infection of H9C2 cells. The cells were divided into four groups: the normal control (NC) group, the ischemia-reperfusion (IR) group, the SIRT3 overexpression +IR (S+IR) group and the SIRT3 overexpression +IR+ SIRT3 inhibitor (S+IR+3-TYP) group. The expressions of SIRT3, Ac-FOXO1, autophagy labeled protein LC3B, Beclin1 and apoptotic proteins Bcl-2 and Bax were detected by Western blot assay. The apoptosis rate was detected by flow cytometry. The level of lactate dehydrogenase (LDH) in the supernatant of cell culture was determined by automatic biochemical analyzer. Results　Compared with the NC group, the expression of SIRT3 was decreased, Ac-FOXO1 was increased, the expressions of autophagy related proteins Beclin1 and LC3B were increased, the expression of anti-apoptotic protein Bcl-2 was decreased, the expression of pro-apoptotic protein Bax was increased, LDH release and cell apoptosis rate were increased in the IR group (P＜0.05). Compared with the IR group, the expression of SIRT3 increased, Beclin1, LC3B and Bcl-2 were increased, the expressions of Ac-FOXO1, pro-apoptotic protein Bax were decreased, LDH release and cell apoptosis rate were decreased in the S+IR group (P＜0.05). After adding SIRT3 inhibitor 3-TYP, there was no significant change in SIRT3 expression compared with that of the S+IR group, but the expression of Ac-FOXO1 was increased, the expressions of Beclin1 and LC3B were decreased, the expression of anti-apoptotic protein Bcl-2 was decreased, the expression of pro-apoptotic protein Bax was increased, LDH release and cell apoptosis rate were increased (P＜0.05). Conclusion　The activation of SIRT3-FOXO1 pathway can increase autophagy level of H9C2 cells, and has a protective effect on myocardial ischemia reperfusion injury.

Key words: myocytes, cardiac, reperfusion injury, autophagy, apoptosis, forkhead box protein O1, silent mating type information regulator 2 homolog 3, H9C2 cell