Rno-miR-30c-5p 通过激活 NF-κB 通路和靶向 SIRT1 促进大鼠心肌缺血再灌注损伤。
Rno-microRNA-30c-5p promotes myocardial ischemia reperfusion injury in rats through activating NF-κB pathway and targeting SIRT1.
机构信息
Department of Cardiology, Luoyang Central Hospital Affiliated to Zhengzhou University, No. 288, Zhongzhou Middle Road, Luoyang City, 471000, Henan Province, China.
Department of Orthopedics, Luoyang Central Hospital Affiliated to Zhengzhou University, No. 288, Zhongzhou Middle Road, Luoyang City, 471000, Henan Province, China.
出版信息
BMC Cardiovasc Disord. 2020 May 20;20(1):240. doi: 10.1186/s12872-020-01520-2.
BACKGROUND
This study aimed to investigate the regulatory effect of rno-microRNA-30c-5p (rno-miR-30c-5p) on myocardial ischemia reperfusion (IR) injury in rats and the underlying molecular mechanisms.
METHODS
A rat model of myocardial IR injury was established. The infarct size was detected by 2,3,5-triphenyltetrazolium chloride staining. The pathologic changes of myocardial tissues were detected by hematoxylin-eosin staining. The apoptosis of myocardial cells was measured by TUNEL staining and flow cytometry. The mRNA expression of rno-miR-30c-5p and Sirtuin 1 (SIRT1) was detected by quantitative real-time PCR. The levels of IL-1β, IL-6 and TNF-α were detected by enzyme linked immunosorbent assay. The protein expression of Bax, Bcl-2, caspase-3, p-IκBα, IκBα, p-NF-κB p65, NF-κB p65 and SIRT1 was detected by Western blot. The interaction between rno-miR-30c-5p and SIRT1 was predicted by TargetScan, and further identified by dual luciferase reporter gene and RNA immunoprecipitation assay.
RESULTS
The myocardial IR injury model was successfully established in rats. IR induced the myocardial injury in rats and increased the expression of rno-miR-30c-5p. Overexpression of rno-miR-30c-5p enhanced the inflammation, promoted the apoptosis, and activated NF-κB pathway in IR myocardial cells. SIRT1 was the target gene of rno-miR-30c-5p. Silencing of SIRT1 reversed the effects of rno-miR-30c-5p inhibitor on the apoptosis and NF-κB pathway in IR myocardial cells.
CONCLUSIONS
Rno-miR-30c-5p promoted the myocardial IR injury in rats through activating NF-κB pathway and down-regulating SIRT1.
背景
本研究旨在探讨微小 RNA-30c-5p(rno-miR-30c-5p)对大鼠心肌缺血再灌注(IR)损伤的调控作用及其潜在的分子机制。
方法
建立大鼠心肌 IR 损伤模型。采用 2,3,5-三苯基四氮唑氯化物染色检测梗死面积。苏木精-伊红染色检测心肌组织病理变化。TUNEL 染色和流式细胞术检测心肌细胞凋亡。实时定量 PCR 检测 rno-miR-30c-5p 和 Sirtuin 1(SIRT1)的 mRNA 表达。酶联免疫吸附试验检测白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)水平。Western blot 检测 Bax、Bcl-2、caspase-3、p-IκBα、IκBα、p-NF-κB p65、NF-κB p65 和 SIRT1 的蛋白表达。通过 TargetScan 预测 rno-miR-30c-5p 与 SIRT1 的相互作用，并用双荧光素酶报告基因和 RNA 免疫沉淀实验进一步验证。
结果
成功建立了大鼠心肌 IR 损伤模型。IR 诱导大鼠心肌损伤，上调 rno-miR-30c-5p 的表达。过表达 rno-miR-30c-5p 增强了 IR 心肌细胞的炎症反应，促进了细胞凋亡，并激活了 NF-κB 通路。SIRT1 是 rno-miR-30c-5p 的靶基因。沉默 SIRT1 逆转了 rno-miR-30c-5p 抑制剂对 IR 心肌细胞凋亡和 NF-κB 通路的影响。
结论
rno-miR-30c-5p 通过激活 NF-κB 通路和下调 SIRT1 促进大鼠心肌 IR 损伤。
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