| 王晓景,张明星,徐 超,管倩倩,曹艺敏,陈小亮.miR-152靶向AT1R对AngⅡ诱导的心肌成纤维细胞增殖及胶原合成的影响[J].,2019,19(16):3055-3061 |
| miR-152靶向AT1R对AngⅡ诱导的心肌成纤维细胞增殖及胶原合成的影响 |
| Effect of miR-152-Targeted AT1R on AngII-induced Proliferation and Collagen Synthesis of Cardiac Fibroblasts |
| 投稿时间：2018-11-23 修订日期：2018-12-18 |
| DOI：10.13241/j.cnki.pmb.2019.16.010 |
| 中文关键词: miR-152 AT1R AngⅡ 心肌纤维化 胶原合成 |
| 英文关键词: miR-152 AT1R AngII Myocardial fibrosis Collagen synthesis |
| 基金项目:国家自然科学基金项目(81102437）；郴州市科技局项目(czkj2016060) |
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| 中文摘要: |
| 摘要 目的：探讨微小RNA-152(miR-152)靶向血管紧张素受体(AT1R)对血管紧张素Ⅱ(AngⅡ)诱导的大鼠心肌成纤维细胞增殖及胶原合成的影响。方法：采用1 μmol/L AngⅡ刺激体外培养的大鼠心肌成纤维细胞，通过噻唑蓝(MTT)法检测细胞增殖情况，蛋白免疫印迹(Western blot)检测成纤维细胞中胶原蛋白I(Collagen I)、胶原蛋白I(Collagen III)以及AT1R蛋白表达的影响，实时荧光定量聚合酶链式反应(qRT-PCR)检测成纤维细胞中miR-152和AT1R mRNA的表达。在AngⅡ诱导的心肌成纤维细胞中分别转染miR-152 mimic和mimic control、AT1R siRNA和siRNA control以及共转染miR-152 mimic和AT1R过表达载体，以同样的方法检测细胞增殖和胶原合成情况。双荧光素酶报告基因实验检测miR-152和AT1R靶向结合关系。结果：AngⅡ刺激能够促进心肌成纤维细胞增殖，上调成纤维细胞中胶原蛋白Collagen I和Collagen III的表达，同时能够抑制miR-152的表达，促进AT1R mRNA和蛋白的表达。在AngⅡ诱导的心肌成纤维细胞中，过表达miR-152或沉默AT1R均能够上调细胞增殖活力，促进胶原合成。双荧光素酶报告基因实验检测结果显示AT1R是miR-152靶基因，miR-152能够负向调控AT1R的表达。在AngⅡ诱导的心肌成纤维细胞中，同时过表达miR-152和AT1R能够逆转单独过表达miR-152导致的细胞增殖抑制作用，回调胶原蛋白Collagen I和Collagen III合成抑制作用。结论：miR-152能够抑制AngⅡ诱导的心肌成纤维细胞增殖和胶原合成，其作用机制可能是通过靶向AT1R的表达实现的。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect of microRNA-152 (miR-152)-targeted angiotensin receptor (AT1R) on angiotensin II (AngII)-induced rat cardiac fibroblast proliferation and collagen synthesis. Methods: Rat cardiac fibroblasts were cultured in vitro and stimulated with 1 μmol/L AngII. The proliferation of fibroblast was detected by MTT assay, the expression of collagen I (Collagen I), collagen I (Collagen III) and AT1R protein in the fibroblasts were detected by western blotting, real-time quantitative polymerase chain reaction (qRT-PCR) was performed to detect the expression of miR-152 and AT1R mRNA in the fibroblasts. MiR-152 mimic and mimic control, AT1R siRNA and siRNA control were transfected and co-transfected with miR-152 mimic and AT1R overexpression vectors in AngII-induced cardiac fibroblasts, respectively, the cell proliferation and collagen synthesis were detected in the same manner. The dual luciferase reporter gene assay was used to detect the targeted binding relationship of miR-152 and AT1R. Results: AngII stimulation can promote the proliferation of cardiac fibroblasts, up-regulate the expression of collagen Collagen I and Collagen III in the fibroblasts, and inhibit the expression of miR-152 and promote the expression of AT1R mRNA and protein. Overexpression of miR-152 or silencing AT1R in AngII-induced cardiac fibroblasts can upregulate cell proliferation and promote collagen synthesis. The results of the dual luciferase reporter gene assay showed that AT1R is a miR-152 target gene, and miR-152 can negatively regulate the expression of AT1R. Simultaneous overexpression of miR-152 and AT1R in AngII-induced cardiac fibroblasts can reverse the inhibition of cell proliferation induced by overexpression of miR-152 alone, and up-regulate the inhibition of collagen Collagen I and Collagen III synthesis. Conclusion: miR-152 can inhibit the proliferation and collagen synthesis of cardiac fibroblasts induced by AngII, and its mechanism may be related to target the expression of AT1R. |
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