Department of Cardiology, Affiliated Hospital of Guilin Medical University, No.20, Lequn Road, Guilin, 541001, Guangxi Zhuang Autonomous Region, People's Republic of China.
Inflamm Res. 2021 Dec;70(10-12):1089-1100. doi: 10.1007/s00011-021-01508-4. Epub 2021 Oct 11.
We aim to explore the molecular mechanism of myocardial ischemia-reperfusion injury (MIRI).
The H9C2 cells were cultured under hypoxia/reoxygenation (H/R) condition to induce myocardial injury in vitro. The expression of miR-451-3p and MAP1LC3B was detected by RT-qPCR. Dual-luciferase reporter assay and RNA pull-down assay were performed to examine the relationship between microRNA (miR)-451-3p and MAP1LC3B. CCK8 was used to test cell viability. The level of LDH and CK was evaluated via ELISA. Immunofluorescence assay and flow cytometry were applied to detect autophagy and apoptosis, respectively. Autophagy-related protein expressions were determined by western blotting. Furthermore, an in vivo rat model of MIRI was established by subjection to 30 min ischemia and subsequently 24 h reperfusion for validation of the role of miR-451-3p in regulating MIRI in vivo.
miR-451-3p was down-regulated in MIRI, and miR-451-3p mimics transfection alleviated autophagy and apoptosis induced by MIRI. miR-451-3p could target MAP1LC3B directly. Co-transfection miR-451-3p mimics and pcDNA 3.1 MAP1LC3B curbed the protected effects of miR-451-3p mimics on MIRI.
miR-451-3p played a protective role in MIRI via inhibiting MAP1LC3B-mediated autophagy, which may provide new molecular targets for the treatment of MIRI and further improves the clinical outcomes of heart diseases.
我们旨在探讨心肌缺血再灌注损伤（MIRI）的分子机制。
在体外培养 H9C2 细胞，使其处于缺氧/复氧（H/R）条件下，以诱导心肌损伤。通过 RT-qPCR 检测 miR-451-3p 和 MAP1LC3B 的表达。通过双荧光素酶报告基因检测和 RNA 下拉实验，研究微 RNA（miR）-451-3p 与 MAP1LC3B 之间的关系。CCK8 用于检测细胞活力。通过 ELISA 评估 LDH 和 CK 的水平。通过免疫荧光和流式细胞术分别检测自噬和凋亡。通过 Western blot 检测自噬相关蛋白的表达。此外，通过 30 分钟缺血和随后 24 小时再灌注建立大鼠 MIRI 体内模型，以验证 miR-451-3p 在体内调节 MIRI 的作用。
miR-451-3p 在 MIRI 中下调，miR-451-3p 模拟物转染可减轻 MIRI 诱导的自噬和凋亡。miR-451-3p 可以直接靶向 MAP1LC3B。共转染 miR-451-3p 模拟物和 pcDNA 3.1 MAP1LC3B 抑制了 miR-451-3p 模拟物对 MIRI 的保护作用。
miR-451-3p 通过抑制 MAP1LC3B 介导的自噬，在 MIRI 中发挥保护作用，这可能为 MIRI 的治疗提供新的分子靶点，并进一步改善心脏疾病的临床转归。
