Furin基因对高血压心肌梗死细胞增殖、迁移以及作用机制的研究

The study of Furin gene on the proliferation,migration and mechanism of hypertensive myocardial infarction cells

目的初步探索Furin与心肌成纤维细胞增殖、迁移的关系以及作用机制。方法 RNAi技术沉默心肌成纤维细胞中Furin基因的表达,分为空白对照组、阴性对照组(转染siRNA-NC)和Furin干扰组(转染siRNA Furin)。四甲基偶氮唑(MTT)实验检测细胞增殖的情况,Transwell法检验细胞的迁移能力,采用荧光定量PCR(RT-PCR)和免疫印迹法(Western blot)检测Furin、α-平滑肌细胞肌动蛋白(α-SMA)、Ⅰ型胶原(Col-1)mRNA、蛋白水平。结果 Furin干扰组心肌成纤维细胞中Furin mRNA、蛋白的表达量均显著低于空白对照组(P<0.05),阴性对照组与空白对照组差异无显著性(P>0.05)。沉默Furin表达显著抑制细胞的增殖和迁移(P<0.05),显著下调Col-1、α-SMA的表达水平(P<0.05),阴性对照组细胞增殖、迁移与空白对照组差异无显著性(P>0.05)。结论沉默Furin能通过抑制心肌成纤维细胞增殖、迁移、胶原合成,增加细胞外基质分解和抑制细胞外基质沉积,降低心肌梗死过程中的心肌纤维化过程。

Objective To explore the relationship between Furin and proliferation,migration of cardiac fibroblasts and its mechanism.Methods RNAi was transfected into cardiac fibroblasts to silence the expression of Furin gene and were divided into control group,negative control group（ transfected with siRNA-NC） and Furin interfected group（ transfected with siRNA Furin）. The proliferation of cardiac fibroblasts cells was measured by MTT assay and the migration was analyzed by Transwell. The mRNA and protein levels of Furin,α-Smooth muscle cell actin（ α-SMA）,Collagen-1（ Col-1） were examined by RT-PCR and Western blotting. Results The expression levels of Furin mRNA and protein in Furin interfected group were significantly lower than those in the blank control group（ P〈0. 05）. Knockdown of Furin expression had the significantly effects on inhibiting cell proliferation and migration（ P〈0. 05）,and decreased the protein levels of Col-1 and α-SMA（ P〈0. 05）. Conclusion Silencing of Furin can reduce myocardial fibrosis during myocardial infarction by inhibiting the proliferation,migration,collagen synthesis,extracellular matrix decomposition and inhibiting the extracellular matrix deposition of myocardial fibroblasts.