微小RNA-155通过靶向缺氧诱导因子1α调控心肌纤维化

周永峰; 雷荣; 杜鹏荣; 刘镇; 李妮妮

doi:10.12125/j.chj.201909065

微小RNA-155通过靶向缺氧诱导因子1α调控心肌纤维化

MicroRNA-155 regulates myocardial fibrosis-related gene expression by targeting hypoxia-inducible factor 1α

摘要

摘要:
目的探讨微小RNA（miR）-155对心肌成纤维细胞（cardiac fibroblasts, CFs）中纤维化相关基因表达的作用及其机制。
方法实验分为对照组、血管紧张素(Ang)Ⅱ组、Scramble组、miR-155 mimic组、AngⅡ+ Scramble组、AngⅡ+ miR-155 mimic组、AngⅡ+ HIF-1α siRNA组。AngⅡ刺激原代分离培养的小鼠CFs，模拟心肌纤维化过程中CFs活化过程，采用PCR及Western blot检测各组miR-155、1型胶原（Col 1）、3型胶原（Col 3）、α-平滑肌肌动蛋白（α-SMA）的表达；采用双荧光素酶报告基因实验检测miR-155与缺氧诱导因子（HIF）-1α 3’端非编码区（3’-UTR）的结合作用。
结果 AngⅡ刺激可显著促进小鼠CFs中纤维化相关基因的表达，同时降低miR-155的水平(P <0.05)；过表达miR-155可显著降低AngⅡ诱导的小鼠CFs中纤维化相关基因的表达（P <0.05）；HIF-1α受到了miR-155的负性调控，并能同miR-155结合；miR-155 mimic和HIF-1α siRNA能一致性的降低AngⅡ诱导的小鼠CFs中纤维化相关蛋白的表达。
结论 miR-155能抑制AngⅡ诱导的CFs纤维化，进而抑制心肌纤维化；HIF-1α是miR-155的靶基因，并介导了miR-155发挥抑制AngⅡ诱导的CFs纤维化的过程。

Abstract:
AIM To investigate the effect and mechanism of microRNA-155 (miR-155) on the expressions of fibrosis-related genes in cardiac fibroblasts (CFs).
METHODS The mice were divided into 7 groups: control group, AngII group, Scramble group, miR-155 mimic group, AngII + Scramble group, AngII + miR-155 mimic group, and AngII + HIF-1α siRNA group. Angiotensin II (AngII) stimulated mouse CFs to simulate the activation of CFs during myocardial fibrosis. PCR and Western blot were used to detect the expressions of miR-155, type 1 collagen (Col 1), type 3 collagen (Col 3) and α-smooth muscle actin (α-SMA). Combination of miR-155 and 3’-untranslated region of hypoxia-inducible factor 1α (HIF-1α) was detected by dual luciferase reporter assay.
RESULTS AngII stimulation significantly promoted the expression of fibrosis-related genes in mouse CFs and decreased the level of miR-155. Over-expression of miR-155 significantly decreased the expression of fibrosis-related genes in mouse CFs induced by AngII. HIF-1α was negatively regulated by miR-155 and could bind to miR-155. miR-155 mimic and HIF-1α siRNA consistently reduced the expressions of fibrosis-associated proteins in mouse CFs.
CONCLUSION miR-155 can inhibit myocardial fibrosis. HIF-1α is a target gene of miR-155 and is involved in the process of miR-155 in inhibiting the expressions of CFs-related proteins and inhibiting myocardial fibrosis.

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