miRNA-10a对小鼠肝纤维化细胞增殖及TGFβ1/Smads信号转导通路表达的影响

Effects of miRNA-10a on the proliferation of hepatic fibrosis cells and the expression of TGF beta 1/Smads signal transduction pathway in mice

目的通过观察肝纤维化(hepatic fibrosis,HF)小鼠肝组织TGFβ1/Smads信号转导通路的表达与HF进展的关系,探讨微小RNA(microRNA,miRNA)-10a对于HF的作用机制。方法选用9周龄健康雄性小鼠(C57BL6/J)40只,按照数字表法随机分为对照组和HF模型组(观察组),对照组生理盐水5μl/g腹腔注射,每周2次,注射8周;观察组10%CCL4橄榄油5μl/g腹腔注射,每周2次,注射8周,制作小鼠HF模型。RT-PCR法检测HF细胞中miRNA-10a表达后,将观察组HF细胞进行培养及miRNA-10a模拟物转染(转染组),CCK-8法检测HF细胞增殖能力,Western blotting检测HF细胞中TGFβ1、Smad7的表达水平。结果与对照组比较,观察组miRNA-10a表达水平明显增加(P<0.05);与对照组比较,转染组肝细胞miRNA-10a表达水平明显降低(P<0.05);与对照组相比,低表达miRNA-10a明显降低观察组TGFβ1的表达水平,提高Smad7的表达水平(均P<0.05)。结论小鼠HF细胞中低表达miRNA-10a,转染miRNA-10a模拟物可明显促进小鼠HF细胞增殖,其作用机制是通过miRNA-10a调控TGFβ1/Smads信号转导通路促进小鼠HF。

Objective To investigate the possible mechanism involved in the effect of micro RNA-10a（ miRNA） on hepatic fi-brosis （HF） , through the observation on the expression of TGFβ1/Smads signal transduction pathway in mice with HF and its correla-tion with the development of hepatic fibrosis. Methods Forty healthy male mice with an age of 9 weeks （ C57BL6/J） were randomly divided into the control group and the HF model group （or the observation group）. The animals in the control group were given normal saline abdominally at a dosage of 5 μ1 /g , twice a week, for a succession of 8 weeks, while the animals in the observation group received 10% CCL4 olive oil also at a dosage of 5 μl / g with the same frequency and for the same length of time to develop the model of hepatic fibrosis. The expression of miRNA-lOa was detected by RT-PCR. HF cells were cultured in the animals of the observation group and miRNA-10a mimics were transfected （the transfection group）. CCK-8 method was used to detect the proliferation of HF cells, and the expression levels of TGFβ1 and Smad 7 in the HF cells were detected by Western-blotting. Results As compared with that of the con-trol group, the expression level of miRNA-lOa in the observation group was increased significantly （P 〈0. 05） ; and as compared with that of the control group, the expression level of miRNA-10a in the transfection group were significantly decreased （P 〈0. 05） ; and fur-thermore ,as compared with that of the control group, the lower expression of miRNA-10a significantly decreased the expression level of TGFβ1 and increased the expression level of Smad 7（ P 〈 0. 05 ） . Conclusion The expression of miRNA-10a in the HF cells was low-er, and the transfected miRNA-10a mimics could significantly promote the proliferation of HF cells, the possible mechanism of which might be associated with the regulation of TGFβ1/Smads signal transduction pathway in HF promotion.]