摘要
目的:探讨mi croRNA-200c(miR-200c)对人瘢痕疙瘩成纤维细胞增殖及胶原合成的影响,并阐明其涉及的TGF-β/Smad通路机制。方法:将mi R-200c mi mics用oligofectami脂质体转染经TGF-β1诱导的人瘢痕疙瘩成纤维细胞,Cell Counting Kit-8(CCK-8)法测细胞增殖变化;3H-脯氨酸掺入法测胶原蛋白水平的变化。相关蛋白表达变化和TGF-β1分泌表达变化分别用Western blot和ELISA法检测。结果:mi R-200c明显抑制经TGF-β1诱导的人瘢痕疙瘩纤维细胞的增殖和胶原合成;mi R-200c能明显减低磷酸化Smad2和Smad3的蛋白表达水平及抑制博莱霉素诱导的TGF-β1分泌。结论:miR-200c能明显抑制人瘢痕疙瘩成纤维细胞增殖及胶原合成,其机制可能与抑制TGF-β/Smad通路相关。
Objective To investigate the effects of microRNA-200c（miR-200c） on cell proliferation and collagen synthesis in human keloid fibroblasts and further elucidate its action mechanisms involved in TGF-β/Smad pathway.Methods miR-200c mimics were transfected keloid fibroblasts by oligofectami package.Cell proliferation was studied by Cell Counting Kit-8 assay（CCK-8）.Collagen synthesis was measured by 3H-proline incorporation assay.Proteins expression and TGF-β1 in keloid fibroblasts were investigated by western blot and Enzyme-linked immunosorbent assay,respectively.Results miR-200c significantly inhibited cell proliferation and collagen synthesis induced by TGF-β1 in keloid fibroblasts.The phosphorylation of Smad2 and Smad3 were markedly reduced by miR-200c treatment;miR-200c inhibited the bleomycin-induced elevation of TGF-b1 expression in the cell.Conclusion miR-200c exhibits inhibitory effects on cell proliferation and collagen synthesis in keloid fibroblasts,and its action mechanisms may involve in TGF-β/Smad pathway.
出处
《中国美容医学》
CAS
2012年第9期1539-1542,共4页
Chinese Journal of Aesthetic Medicine
