Affiliated Hospital of Jiaxing University, The First Hospital of Jiaxing, Jiaxing 314000, Zhejiang, China.
Contrast Media Mol Imaging. 2022 Mar 19;2022:7021200. doi: 10.1155/2022/7021200. eCollection 2022.
To investigate the protective effect of miR-542-3p on cardiomyocyte injury and related mechanisms.
A cardiomyocyte hypoxia/reoxygenation model was established. The expression levels of miR-542-3p and PDE4D were detected using qRT-PCR; the luciferase reporter assay system was used to detect the targeting relationship between miR-542-3p and PDE4D; overexpressing miR-542-3p was transfected into cardiomyocytes, and ROS release was detected by immunofluorescence while cellular apoptosis was detected by TUNEL; and the western blot assay was applied to detect the expression of PDE4D, phosphorylated protein kinase A (p-PKA), and phosphorylated cyclic adenosine monophosphate (cAMP) response element-binding protein (p-CREB).
Compared with the control group, the miR-542-3p expression level was decreased and the PDE4D expression level was increased in the cardiomyocyte hypoxia/reoxygenation model group. The dual-luciferase reporter assay system confirmed that miR-542-3p could target and regulate PDE4D; the transfection with cardiomyocytes using the overexpressing miR-542-3p could downregulate PDE4D expression, attenuate ROS release during cardiomyocyte injury, and reduce cellular apoptosis rate, while upregulating the expression of p-PKA and p-CREB.
The miR-542-3p can negatively regulate PDE4D protein expression and attenuate cardiomyocyte injury through a mechanism related to the activation of the cAMP/PKA signaling pathway.
探讨 miR-542-3p 对心肌细胞损伤的保护作用及其相关机制。
建立心肌细胞缺氧/复氧模型。采用 qRT-PCR 检测 miR-542-3p 和 PDE4D 的表达水平；采用荧光素酶报告基因检测系统检测 miR-542-3p 与 PDE4D 的靶向关系；转染 miR-542-3p 过表达质粒入心肌细胞，免疫荧光法检测 ROS 释放，TUNEL 法检测细胞凋亡；Western blot 法检测 PDE4D、磷酸化蛋白激酶 A（p-PKA）和磷酸化环磷酸腺苷反应元件结合蛋白（p-CREB）的表达。
与对照组相比，心肌细胞缺氧/复氧模型组 miR-542-3p 的表达水平降低，PDE4D 的表达水平升高。双荧光素酶报告基因检测系统证实 miR-542-3p 可靶向调控 PDE4D；转染 miR-542-3p 过表达质粒入心肌细胞可下调 PDE4D 表达，减轻心肌细胞损伤时 ROS 的释放，降低细胞凋亡率，同时上调 p-PKA 和 p-CREB 的表达。
miR-542-3p 可通过负向调控 PDE4D 蛋白表达，激活 cAMP/PKA 信号通路，减轻心肌细胞损伤。
