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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mi R-24的表达水平。在心脏成纤维细胞中转染mi R-24 mimics、mimics control、mi R-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mi R-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mi R-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mi R-24表达上调。mi R-24 mimics组中Col-1、α-SMA的表达水平明显低于mimics control组(P<0.01)。mi R-24 mimics组中细胞OD值较mimics control组显著降低(P<0.01),mi R-24 inhibitors组中细胞OD值较inhibitors control组显著升高(P<0.01)。mi R-24 mimics组、mimics control组、mi R-24 inhibitors组、inhibitors control组细胞凋亡无显著性差异(P>0.05)。mi R-24 mimics组细胞迁移数目明显低于mimics control组,差异显著(P<0.01),mi R-24 inhibitors组细胞迁移数目高于inhibitors control组,差异显著(P<0.05)。mi R-24 mimics组细胞中Furin蛋白和m RNA水平明显降低,野生型Furin和mi R-24 mimics共转染的细胞中荧光素酶活性最低。结论:mi R-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。
Objective: To investigate the effect and mechanism of miR-24 on cardiac fibroblast growth and migration. Methods: The expression of mi R-24 in cardiomyocytes and cardiac fibroblasts was detected by RT-PCR. After transfection with mi R-24 mimics, mimics control, mi R-24 inhibitors and inhibitor control in cardiac fibroblasts, the expression of Col-1 and α-SMA was detected by Western blot. The proliferation and flow cytometry Cytometry was used to detect apoptosis. Transwell chamber was used to detect cell migration. The target gene of mi R-24 was predicted by target gene prediction library, and the target gene was identified by luciferase. The expression of Furin in transfected cells was detected by RT-PCR and Western blot. Results: The expression of mi R-24 in cardiac fibroblasts HEH2 was significantly different from that of H9C2 (P <0.01). The expression of mi R-24 in cardiac fibroblasts was up-regulated. The expression of Col-1 and α-SMA in mi R-24 mimics group was significantly lower than that in mimics control group (P <0.01). The OD value of cells in mi R-24 mimics group was significantly lower than that in mimics control group (P <0.01). The OD value of cells in mi R-24 inhibitors group was significantly higher than that in inhibitors control group (P <0.01). There was no significant difference in mi R-24 mimics group, mimics control group, mi R-24 inhibitor group and inhibitor control group (P> 0.05). The number of cell migration in the mi R-24 mimics group was significantly lower than that in the mimics control group (P <0.01), and the number of cell migration in the mi R-24 inhibitor group was significantly higher than that in the inhibitor control group (P <0.05). Furin protein and m RNA levels were significantly decreased in the mi R-24 mimics group, and luciferase activity was the lowest in wild-type Furin and mi R-24 mimics co-transfected cells. Conclusion: mi R-24 is overexpressed in cardiac fibroblasts. The target gene Furin inhibits the synthesis of Col-1 and α-SMA from cardiac fibroblasts and inhibits the proliferation and migration of cardiac fibroblasts.