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

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(C57 BL6/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5 μl/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.

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

Objective To investigate the possible mechanism involved in the effect of micro RNA-10a(miRNA) on hepatic fibrosis (HF),through the observation on the expression of TGFβ1/Smads signal transduction pathway in mice with HF and its correlation 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 μl/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-10a 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 control group,the expression level of miRNA-10a 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 furthermore,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 lower,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.