沉默信息调节因子1对急性心肌梗死小鼠的作用研究

doi:10.16150/j.1671-2870.2018.06.009

摘要/Abstract

摘要： 目的：探讨沉默信息调节因子1 (Sirtuin1, Sirt1) 对急性心肌梗死（简称心梗）小鼠的作用及其机制。方法：通过结扎小鼠心脏冠状动脉左前降支构建急性心梗小鼠模型, 并于心梗后不同时间（1、3、5、7 d）收集梗死区及梗周区域组织（以假手术组心肌组织为对照）, 利用实时定量PCR检测Sirt1表达情况。另构建小鼠心梗模型, 在心梗手术结扎成功的同时, 用微量注射针在心梗组织周围注射Sirt1过表达慢病毒（以阴性对照慢病毒点注射组为对照）, 于术后7 d通过2, 3, 5-氯化三苯基四氮唑（2, 3, 5-triphenyltetrazolium chloride, TTC）染色法检测心梗面积。体外培养小鼠原代心肌细胞, 并于缺氧（2% O₂）条件下, 使用不同浓度（25、50及100 μmol/L）白藜芦醇刺激小鼠原代心肌细胞, 采用蛋白印迹法检测Sirt1和半胱氨酸蛋白酶3剪切体的表达水平, 利用细胞凋亡检测试剂（TUNEL）检测细胞凋亡情况。结果：发现在梗死后1~7 d中, 梗死区及梗周区组织中的Sirt1 mRNA表达水平呈显著下降趋势（P<0.05）。同时, 相较于阴性对照慢病毒注射组, Sirt1过表达慢病毒注射组可显著性减小梗死面积[（15.68±3.28）%比（29.50±5.56）%, P<0.05]。随着白藜芦醇刺激浓度的增加, Sirt1蛋白表达水平呈剂量依赖型升高, 而半胱氨酸蛋白酶3剪切体蛋白表达水平呈下降趋势;心肌细胞的凋亡也随刺激浓度的增加逐渐减少。结论：Sirt1在急性心梗组织中表达下降, 过表达Sirt1可减少急性心梗后的心梗面积。

关键词: 沉默信息调节因子1, 白藜芦醇, 梗死, 急性心梗

Abstract: Objective: To investigate the effect of silencing information regulator factor 1 (Sirtuin1, Sirt1) on acute myocardial infarction in mice and its mechanism. Methods: Model of acute myocardial infarction in mice were constructed by ligating the left anterior descending coronary artery. The infarcted and peripheral tissue were collected at different time after myocardial infarction (1, 3, 5 and 7 days) (the myocardial tissue of the sham operation group was taken as the control), and the expression of Sirt1 was detected by RT-PCR. In addition, in another series of model of myocardial infarction in mice, when successful ligation of coronary artery was established the Sirt1 overexpression lentivirus was injected into the tissue surrounding the infarction by microinjection (negative group of lentiviral injection served as the control). Myocardial infarction area was detected by 2, 3, 5-triphenyltetrazolium chloride (TTC) staining seven days after operation. Mouse primary myocardial cells were cultured in vitro under hypoxia (2%O₂), and resveratrol (Res) of different concentrations (25, 50 and 100 μmol/L) were used to stimulate the primary myocardial cells. The expression levels of Sirt1 and cysteine protease-3 shear bodies (cleaved caspase3) were detected by Western blotting, and cell apoptosis was detected by TUNEL method. Results: mRNA levels of Sirt1 in myocardial infarction and peripheral tissues were significantly decreased 1-7 days after infarction (P<0.05). Meanwhile, compared with the negative control group, the Sirt1 overexpression lentiviral injection group significantly decreased the area of infarction [(15.68±3.28)% vs (29.50±5.56)%, P<0.05)]. As the concentration of resveratrol increased, Sirt1 protein expression increased in a dose-dependent manner, while cleaved caspase-3 protein expression was decreased. The apoptosis of myocardial cells decreased gradually with the increase of concentration of resveratrol stimulation. Conclusions: The expression of Sirt1 declines in acute myocardial infarction. Overexpression of Sirt1 reduces the myocardial infarction area after acute myocardial infarction.

Key words: Sirtuin1, Resveratrol, Infarction, Acute myocardial infarction

中图分类号:

R542.2⁺2

陈亚芬, 陈媛媛, 吴丽苹, 薛琪琪, 杨克, 陆林, 曹久妹. 沉默信息调节因子1对急性心肌梗死小鼠的作用研究[J]. 诊断学理论与实践, 2018, 17(06): 670-675.

CHEN Yafen, CHEN Yuanyuan, WU Liping, XUE Qiqi, YANG Ke, LU Lin, CAO Jiumei. Effect of silencing information regulator factor 1 on mice with acute myocardial infarction[J]. Journal of Diagnostics Concepts & Practice, 2018, 17(06): 670-675.

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