Zhao Yintao, Sun Jingjing, Liu Yuan, Zheng Lu, Zhang Xiangqin, Yang Haibo
Department of Cardiology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, 450052, China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2020 Sep 10;37(9):980-986. doi: 10.3760/cma.j.cn511374-20190821-00422.
To explore the effect and mechanism of miR-125a-5p targeted regulation of scavenger receptor B1 (Scarb1) gene on anoxia/reoxygenation injury of rat cardiomyocytes.
H9c2 rat cardiomyocytes were randomly divided into blank control group, hypoxia/reoxygenation group, transfection control group and mir-125a-5p transfection group. The expression of miR-125a-5p, cardiomyocyte viability, apoptosis rate, ATP content and the expression of Scarb1, Cyt C, Bax, Bcl-2 and NF-κB signaling pathway related proteins were determined. Target gene of miR-125a-5p was predicted with Targetscan software, and the targeting of miR-125a-5p on Scarb1 was verified by double luciferase reporter gene experiment.
Compared with the blank control group, the expression of miR-125a-5p, Bax, Cyt C and the apoptotic rate of cardiomyocytes in the hypoxia/reoxygenation group were significantly increased (P<0.05), while the expression of Scarb1, Bcl-2 and the content of ATP were significantly decreased (P<0.05). Compared with the control group, the situation of mir-125a-5p transfection group was just the opposite. Double luciferase reporter gene experiment has confirmed Scarb1 to be the target of miR-125a-5p. Hypoxia/reoxygenation can promote the expression of NF-κB p65, C-myc and Cyclin D1 in cardiomyocytes, while down-regulating the expression of miR-125a-5p can inhibit the expression of such proteins.
Hypoxia/reoxygenation can induce the expression of miR-125a-5p in rat cardiomyocytes. Inhibition of miR-125a-5p can protect cardiomyocytes from hypoxia/reoxygenation by up-regulating the expression of Scarb1. The mechanism may be related to the inhibition of activation of NF-κB signaling pathway.
探讨微小RNA-125a-5p(miR-125a-5p)靶向调控清道夫受体B1(Scarb1)基因对大鼠心肌细胞缺氧/复氧损伤的影响及机制。
将H9c2大鼠心肌细胞随机分为空白对照组、缺氧/复氧组、转染对照组和miR-125a-5p转染组。检测miR-125a-5p的表达、心肌细胞活力、凋亡率、ATP含量以及Scarb1、细胞色素C(Cyt C)、Bax、Bcl-2和核因子κB(NF-κB)信号通路相关蛋白的表达。用Targetscan软件预测miR-125a-5p的靶基因,并通过双荧光素酶报告基因实验验证miR-125a-5p对Scarb1的靶向作用。
与空白对照组比较,缺氧/复氧组miR-125a-5p、Bax、Cyt C的表达及心肌细胞凋亡率明显升高(P<0.05),而Scarb1、BclBcl-2的表达及ATP含量明显降低(P<0.05)。与转染对照组比较,miR-125a-5p转染组情况相反。双荧光素酶报告基因实验证实Scarb1是miR-125a-5p的靶基因。缺氧/复氧可促进心肌细胞中NF-κB p65、C-myc和细胞周期蛋白D1(Cyclin D1)的表达,而下调miR-125a-5p的表达可抑制这些蛋白的表达。
缺氧/复氧可诱导大鼠心肌细胞miR-125a-5p的表达。抑制miR-125a-5p可通过上调Scarb1的表达保护心肌细胞免受缺氧/复氧损伤。其机制可能与抑制NF-κB信号通路的激活有关。