黄芩苷减轻HFLS-RA合成细胞外基质的分子机制研究

Study on the molecular mechanism of baicalin on alleviating synthesis of extracellular matrix of HFLS-RA

目的 研究黄芩苷对类风湿关节炎成纤维样滑膜细胞(HFLS-RA)miR-21、TGF-β1、Smad7及FN的影响,为阐释黄芩苷抗RA滑膜纤维化的分子机制提供依据。方法 采用CCK8法检测黄芩苷对HFLS-RA细胞活性的影响;采用RT-qPCR法测定黄芩苷对HFLS-RA细胞miR-21的影响;采用Western blot技术测定黄芩苷对HFLS-RA细胞TGF-β1、Smad7及FN蛋白表达的影响。结果 CCK8检测结果显示,随着黄芩苷浓度增大和作用时间的延长,黄芩苷对HFLS-RA细胞的生长抑制作用增强,24h、48h的半数抑制浓度(IC50)分别为(243.8±12.43)、(236.5±10.56)μg/ml;黄芩苷100、50及25μg/ml浓度可有效抑制类风湿关节炎成纤维样滑膜细胞miR-21、TGF-β1及FN表达(P<0.05),而促进Smad7表达(P<0.05)。结论 黄芩苷通过下调RA滑膜成纤维细胞的miR-21表达、促进Smad7蛋白表达,进而抑制TGF-β1的表达,从而减轻RA滑膜成纤维细胞过度分泌合成细胞外基质。

Objective To study the effects of Baicalin on miR-21,TGF-β1,Smad7 and FN in fibroblast-like synoviocytes(HFLS-RA)of rheumatoid arthritis(RA),and to elucidate the molecular mechanism of baicalin against RA synovial fibrosis.Methods CCK8 method was used to detect the effect of Baicalin on the activity of HFLS-RA cells,RT-q PCR was used to determine the effect of Baicalin on the on miR-21;Western blot was adopted to check HFLS-RA protein expressions of TGF-β1,Smad7 and FN in HFLS-RA.Results CCK8 result showed that the growth inhibition of HFLS-RA cells was enhanced with the increase of baicalin concentration and the prolongation of time.The half inhibitory concentrations(IC50)of baicalin at 24 h and 48 h were(243.8±12.43),(236.5±10.56)μg/ml respectively;Baicalin at 100,50 and 25μg/ml could effectively inhibit the expression of miR-21 and the protein expressions of TGF-β1 and FN(P<0.05),while the protein expression of Smad7 increased significantly(P<0.05).Conclusion Baicalin can inhibit the expression of TGF-β1 by down-regulating the expression of miR-21 and promoting the expression of Smad7 protein in RA synovial fibroblasts,thereby reducing the excessive synthesis of extracellular matrix of RA synovial fibroblasts.