益肾软坚散含药血清拮抗马兜铃酸对人肾间质成纤维细胞的作用

Antagonistic Effect of Yishen Ruanjian San Contained Serum against Aristolochic Acid in Antagonizing Human Renal Interstitial Fibroblasts

目的研究益肾软坚散含药血清能否干预马兜铃酸 (aristolochicacid ,AA)对人肾间质成纤维细胞 (humanrenalinterstitialfibroblasts,hRIFs)的致细胞外基质 (excellularmatrix ,ECM )蓄积效应。方法用马兜铃酸钠盐 (AA Na ,4 0 μg/ml)加或不加 10 %大鼠含药血清与hRIFs孵育 ,而后检测hRIFs对转化生长因子 β1(transforminggrowthfactor β1,TGF β1)、结缔组织生长因子 (connectivetissuegrowthfactor,CTGF)、纤溶酶原激活物抑制物 1(plasminogenactivatorinhibitor 1,PAI 1)、金属蛋白酶组织抑制物 1(tissueinhibitorofmetalloproteinase 1,TIMP 1)和Ⅰ型胶原 (typeⅠCollagen ,ColⅠ )的mRNA(RT PCR方法 )和蛋白质表达(ELISA或Westernblot方法 )。结果大鼠含药血清能显著下调AA Na刺激后hRIFs对TGF β1、CTGF、TIMP 1和ColⅠmRNA及蛋白质高表达 (P <0 .0 5 ) ,但不影响PAI 1mRNA及蛋白质高表达 (P >0 .0 5 )。结论益肾软坚散含药血清可通过下调hRIFs促ECM合成因子及抗ECM降解因子的表达 。

Objective To study whether Yishen Ruanjian San contained serum (S YRS) could intervene the action of aristolochic acid (AA) in antagonizing human renal interstitial fibroblasts (hRIFs) to induce extracellular matrix (ECM) accumulation. Methods AA Na 40 μg/ml, with or without 10% S YRS, was co cultured with hRIFs, then the hRIFs mRNA of transforming growth factor β 1 (TGF β 1), connective tissue growth factor (CTGF), plasminogen activator inhibitor 1 (PAI 1), tissue inhibitor of metalloproteinase 1 (TIMP 1) and type Ⅰ collagen (ColⅠ) in the cultured cells were detected by RT PCR, and their protein expression monitored with ELISA and Western blot respectively. Results The mRNA and protein expression of all the above mentioned factors were significantly up regulated by AA Na (P<0 05). Excepting PAI 1, the enhanced mRNA and protein expression were significantly down regulated by S YRS (P<0 05). Conclusion S YRS could down regulate the hRIF to promote the expression of ECM synthesis factors and inhibit the ECM degradation factors in hRIFs, so as to antagonize the AA stimulated accumulation of ECM such as Col I.