RESUMEN
Objective 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.
RESUMEN
Objective To investigate the effect of miRNA-200b-specific inhibitor on hepatic stellate cells(HSCs) activation,proliferation,and extracellular matrix production.Methods The miRNA-200b-specific inhibitors were designed,synthesized,and transfected into HSCs with lipofectamine 2000.The supernatant and HSCs were collected after incubation for 48 h.The expression of miR-200b was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR).The expression ofα-smooth muscle actin (oα-SMA) protein in HSCs was detected by Western blotting.The cell proliferation was assayed by methyl thiazolyl tetrazolium (MTT) method.Contents of type Ⅲ procollagen and hyaluronic acid in supernatant were determined by radioimmunoassay.Results Compared to the control group,miRNA-200b expression was decreased in the miRNA-200b inhibitor group by 82％ (P ＜ 0.01),α-SMA protein expression was reduced in the miRNA-200b inhibitor group by (19 ± 3) ％ (P ＜ 0.05),and the activity of HSCs proliferation was reduced by(33 ± 5)％ (P ＜0.01),and the contents of type Ⅲ procollagen and hyaluronic acid in supernatant were reduced in miRNA-200b inhibitor group by (35 ± 4)％ and (31 ± 2)％,respectively(P ＜0.01).Conclusions The miRNA-200b-specific inhibitor could significantly reduce the expression of miRNA-200b,and inhibit HSC proliferation,activation,and extracellular matrix production.