微小RNA-130b在转化生长因子β1诱导的肾小球系膜细胞纤维化中的调控作用

Regulation of miR-130b on glomerular mesangial cell fibrosis induced by transforming growth factor β1

目的观察微小RNA^(-1)30b(miR^(-1)30b)对转化生长因子β1(TGF-β1)蛋白诱导的人肾小球系膜细胞(HMC)纤维化的调控作用。方法用高糖(25mmol·L^(-1))及低糖(5.5 mmol·L^(-1))体外培养人肾小球系膜HMC细胞,miR^(-1)30b高表达慢病毒和TGF-β1作为干预因素,分别设低糖、高糖24和48 h组、TGF-β1(10,30 ng·m L^(-1))24,48 h实验组和慢病毒miR^(-1)30b组、慢病毒miR-NC组。Western blot与细胞免疫荧光技术检测胶原蛋白Ⅰ(colⅠ)的表达;实时荧光定量聚合酶链式反应(qRT-PCR)法检测miR^(-1)30b和colⅠmRNA的表达。结果在TGF-β1诱导下,慢病毒miR^(-1)30b组、慢病毒miR-NC组的miR^(-1)30b表达量分别为6.01±1.09,1.00;colⅠmRNA表达量分别为2.88±0.64,1.00;colⅠ蛋白的表达量分别为2.11±0.22,1.00,组间比较差异有统计学意义(P<0.01)。结论 miR^(-1)30b、colⅠmRNA和蛋白在TGF-β1诱导的人肾小球系膜细胞纤维化过程中慢病毒miR^(-1)30b组较miR-NC组表达均增高,miR^(-1)30b可能与TGF-β1诱导的人肾小球系膜细胞纤维化相关。

Objective To investigate the effect of miR-130 b on the fibrosis of human mesangial cells(HMC） induced by transforming growth factor β 1(TGF-β1）. Methods In vitro HMC cells were cultured with high glucose(25 mmol·L-1） or low glucose(5. 5 mmol·L-1）,treated by miR-130 b high expression of lentivirus or TGF-β1,respectively divided into low glucose,high glucose 24,48 h groups,10,30 ng·m L-1 TGF-β1 24,48 h groups,miR-130 b lentivirus group and miR-130 b-NC group. Western blot and immunofluorescence technique were used to detect the expression of collagen type Ⅰ protein(col Ⅰ）,and the expression of miR-130 b and colⅠ mRNA were detected by real-time polymerase chain reaction(qRT-PCR）. Results Induced by TGF-β1,the expression of miR-130 b in miR-130 b lentivirus group and miR-130 b-NC group were 6. 01 ± 1. 09,1. 00; col Ⅰ mRNA expression were 2. 88 ± 0. 64,1. 00,and col Ⅰ potein expression were2. 11 ± 0. 22,1. 00,all with significant difference(all P〈0. 01）.Conclusion miR-130 b,col Ⅰ mRNA and protein were all highly expressed in the process of mesangial cell fibrosis induced by TGF-β1,and miR-130 b may be related to the induction of mesangial cell fibrosis induced by TGF-β1.