微小RNA-199a对人胚肺成纤维细胞表型转化的抑制作用机制研究

目的 探讨微小RNA-199a(miR-199a)对转化生长因子-β1(TGF-β1)诱导的人胚肺成纤维细胞(HELF)向肌成纤维细胞转化的作用及机制.方法 将细胞分为6组:对照组、模型组(5 ng·mL-1的TGF-β1诱导HELF细胞72 h)、第1转染组(转染150 nmol·L-1的inhibitor NC质粒...

Inhibits effect and mechanism of miR-199a on the phenotypic transformation of human embryonic lung fibroblast cells

Objective To investigate the effect and mechanism of microRNA-199 a(miR-199 a)on the phenotypic transformation of human embryonic lung fibroblast cells(HELFs)induced by transforming growth factor-β1(TGF-β1).Methods The cells were devided into six groups:control group,model group(HELFs induction 5 ng·mL^-1 TGF-β1 for 72 h),trasfected-1 group(transfection 150 nmol·L^-1 inhibitor NC plasmid),trasfected-2 group(transfection 150 nmol·L^-1 miR-199 a inhibitor plasmid),trasfected-3 group(transfection inhibitor NC plasmid+TGF-β1 induction)and trasfected-4 group(transfection miR-199 a inhibitor plasmid+TGF-β1 induction).Real-time quantitative-PCR assay was used to detect the expression(2-△△Ctvalue)of miR-199 a.Western blot was used to detect the expression(gray value)ofα-smooth muscle actin(α-SMA),CollagenⅠ,phosphorylation-serine threonine protein kinase(p-Akt)and phosphorylation-mammalian target of rapamycin(p-m TOR).Results The expression levels of miR-199 a in control group,model group,trasfected-1 group,trasfected-2 group,trasfected-3 group and trasfected-4 group were 0.99±0.00,4.73±0.29,0.93±0.12,0.37±0.01,4.52±0.22 and 0.31±0.08;comparing between model group and control group,the differences was significant(P<0.001).The expression levels ofα-SMA protein in model group,trasfected-3 group and trasfected-4 group were 0.58±0.13,0.58±0.10 and 0.36±0.03;the expression levels of CollagenⅠprotein in the above 3 groups were 0.50±0.05,0.49±0.07 and 0.29±0.04;the expression levels of p-Akt(S473)/Akt protein in the above 3 groups were0.60±0.15,0.59±0.16 and 0.30±0.09;the expression levels of p-m TOR/m TOR protein in the above 3 groups were 0.74±0.02,0.74±0.04 and 0.49±0.06;comparing between trasfected-4 group and trasfected-3 group,the differences of the factors were significant(P<0.05,P<0.01).Conclusion The miR-199 a promotes the transformation of HELF cells into myofibroblasts induced by TGF-β1,and its mechanism may be related to the activation of PI3 K/Akt/m TOR pathway by miR-199 a.