| 赵玉明,李艳星,朱晓锋,吴飞马,李振华,钟声逸,李丽红.miR-24对心脏成纤维细胞生长和迁移的影响及机制研究[J].现代生物医学进展英文版,2017,17(16):3039-3044. |
| miR-24对心脏成纤维细胞生长和迁移的影响及机制研究 |
| Effect and Mechanism of miR-24 on Cardiac Fibroblasts |
| Received:August 31, 2016 Revised:September 20, 2016 |
| DOI：10.13241/j.cnki.pmb.2017.16.010 |
| 中文关键词: 心脏成纤维细胞 miR-24 增殖 迁移 |
| 英文关键词: Cardiac fibroblasts miR-24 Proliferation Migration |
| 基金项目:湖北省卫生厅科研基金项目(JX4B83) |
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| 中文摘要: |
| 摘要 目的：探讨miR-24对心脏成纤维细胞的生长和迁移的影响及机制。方法：采用RT-PCR检测心肌细胞和心脏成纤维细胞中miR-24的表达水平。在心脏成纤维细胞中转染miR-24 mimics、mimics control、miR-24 inhibitors、inhibitors control后，通过Western blot检测细胞中Col-1、α-SMA的表达，MTT检测细胞增殖情况，流式细胞仪检测细胞凋亡情况，Transwell小室检测细胞迁移能力。通过靶基因预测库预测miR-24的靶基因，荧光素酶鉴定靶基因的正确性，并通过RT-PCR和Western blot检测转染后细胞中Furin的表达。结果：心脏成纤维细胞HEH2中miR-24的表达水平与心肌细胞H9C2相比差异显著(P<0.01)，心脏成纤维细胞中miR-24表达上调。miR-24 mimics组中Col-1、α-SMA的表达水平明显低于mimics control组(P<0.01)。miR-24 mimics组中细胞OD值较mimics control组显著降低(P<0.01)，miR-24 inhibitors组中细胞OD值较inhibitors control组显著升高(P<0.01)。miR-24 mimics组、mimics control组、miR-24 inhibitors组、inhibitors control组细胞凋亡无显著性差异(P>0.05)。miR-24 mimics组细胞迁移数目明显低于mimics control组，差异显著(P<0.01)，miR-24 inhibitors组细胞迁移数目高于inhibitors control组，差异显著(P<0.05)。miR-24 mimics组细胞中Furin蛋白和mRNA水平明显降低，野生型Furin和miR-24 mimics共转染的细胞中荧光素酶活性最低。结论：miR-24在心脏成纤维细胞中高表达，通过靶基因Furin抑制心脏成纤维细胞合成Col-1、α-SMA，抑制心脏成纤维细胞增殖和迁移。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the role of miR-24 on the growth and migration of cardiac fibroblasts and its mechanisms. Methods: RT-PCR was used to detect the expression level of miR-24 in myocardial cells and cardiac fibroblasts. miR-24 mimics, mimics control, miR-24 inhibitors, inhibitors control were transfected into the cardiac fibroblasts, then the expression levels of Col-1, α-SMA in cardiac fibroblasts were detected. MTT was used to detect the cell proliferation. Flow cytometry was performed to detect the cell apoptosis. Transwell was used to detect the cell migration. miR-24 target gene was predicted through target gene prediction software. Identification of target was corrected by luciferase gene. The expression level of Furin was detected by RT-PCR and Western blot. Results: The expression levels of miR-24 in cardiac fibroblasts was significantly upregulated compared with the cardiomyocytes (P<0.01). The expression levels of Col-1 and -SMA in mimics miR-24 group were significantly lower than those in control group (P<0.01). The OD value of miR-24 mimics compared to mimics control was significantly decreased (P<0.01). The OD value of miR-24 inhibitors compared to inhibitors control was significantly increased (P<0.01). No significant difference was found in the apoptosis between miR-24 mimics group, mimics control group, miR-24 inhibitors group, inhibitors control group (P>0.05). The number of cell migration in miR-24 mimics group was significantly lower than that of mimics control group (P<0.01), the number of cell migration in miR-24 inhibitors group was higher than that of inhibitors control group (P<0.05). The levels of Furin protein and mRNA in mimics miR-24 group were significantly lower, and the luciferase activity was the lowest in wild type Furin and mimics miR-24 co transfected cells. Conclusion: miR-24 was highly expressed in cardiac fibroblasts, which might inhibit the proliferation and migration of cardiac fibroblasts by inhibiting the synthesis of Col-1 and α-SMA in cardiac fibroblasts by target gene Furin. |
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