摘要目的 探讨MicroRNA-9 (miR-9)对肝星状细胞增殖和胶原合成的影响并研究其相关机制.方法 设计合成miR-9特异性模拟物(miR-9 mimics),将其转入肝星状细胞中,培养48 h后收集细胞,采用实时定量PCR技术(qRT-PCR)验证miR-9 mimics转染细胞中miR-9的变化水平、Western blot检测Ⅰ型胶原、Ⅲ型胶原蛋白表达、噻唑蓝(MTT)法检测细胞增殖情况;通过qRT-PCR和Western blot检测转染miR-9 mimics后其潜在靶点转化生长因子-β1受体2(TGFBR2)的表达水平.结果 与对照组相比,转染miR-9 mimics后,检测肝星状细胞中miR-9表达量明显升高(P＜0.05).过表达miR-9后Ⅰ型胶原、Ⅲ型胶原蛋白分别降低约(44±2)％和(50±3)％(P＜0.05),细胞增殖活性降低约(48±4)％(P＜0.01),TGFBR2的表达水平降低(P＜0.05).结论 过表达miR-9可抑制肝星状细胞胶原合成和细胞增殖,此机制可能是通过下调TGFBR2的表达来实现.

abstractsObjective To investigate the effect of MicroRNA-9 (miR-9) on cell proliferation and collagen synthesis in hepatic stellate cells (HSCs),and to explore the potential mechanism.Methods HSC-T6 cells were cultured and transfected with miR-9 mimics with lipofectamine 2000.After incubation 48 h,the cells were collected and total proteins and RNAs were extracted.The expression of miR-9 was detected by quantitative real time polymerase chain reaction (qRT-PCR).The protein expression of type Ⅰ collagen and type Ⅲ collagen were measured by Western blot.The methyl thiazolyl tetrazolium (MTT) method was used to asses the proliferation of HSC-T6 cells.The expression of transforming growth factor-β1 receptor 2 (TGFBR2) was detected by qRT-PCR and Western blot.Results Compared to the control group,miR-9 expression in HSCs was increased in the miR-9 mimics group (P ＜ 0.05),type Ⅰ and type Ⅲ collagen protein expression was reduced by (44 ± 2) ％ and (50 ± 3) ％ (P ＜ 0.01),respectively.The proliferation activity of HSCs was decreased by (48 ± 4)％ (P ＜ 0.05).The expression of TGFBR2 was inhibited in the miR-9 mimics group.Conclusions Upregulation of miR-9 plays a role on suppressing cell proliferation and collagen synthesis in HSCs.This process might be mediated by downregulation of TGFBR2.