2010•Chinese Journal of Integrated Traditional and Western NephrologyRequires access
Effects of Wenyanghuoxuefang on Aristolochic Acid Stimulated Transdifferentiation of Renal Tubular Epithelial Cells in Vitro
Yunman Wang
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Abstract
Objective:To investigate the effect of transdifferentiation for the aristolochic acid (Aristolochic Acid,AA) on rat proximal tubular epithelial cells (NRK-52E),and antagonistic role of Wenyanghuoxue for transdifferentiation.Methods:(1)To observe the proliferation of NRK-52E by MTT assay under the intention of different concentrations of Wenyanghuoxuefang containing serum.(2) To observe the expression of α-smooth muscle actin protein for NRK-52E under intention of different concentration AA (0,5,10,20,40 μg/ml) by immunocytochemical method.(3) to assay the expression of Col-Ⅰ and regulatory role of Wenyanghuoxuefang by ELISA assay.(4) to assay the expression of TGF-β1,VEGF,ET-1for NRK-52E under Wenyanghuoxuefang containing serum treatment.Results:(1) AA 5 μg/ml,AA 10 μg/ml,AA 20 μg/ml,AA 40 μg/ml can stimulate the NRK-52E expressing α-smooth muscle actin,the group of AA 10 μg/ml have the strongest role of stimulation.(2) After AA (10 μg/ml) stimulating cells at different time points(12、24、48 h),the expression of Col-Ⅰ increasedand showed time-depended trend.(3) After AA stimulating 24 hours,the expression of TGF-β1 mRNA was increased (P0.05),VEGF m RNA was significantly lower (P0.01),ET-1 mRNA was significantly raised (P0.01),Col-Ⅰ was significantly higher (P0.01),for the group of Wenyanghuoxuefang,the expression of TGF-β1 mRNA decreased (P0.05),the expression of VEGF mRNA was significantly raised (P0.01),ET-1 mRNA showed downward trend,the expression of Col-Ⅰ was significantly reduced.Conclusion:(1)Aristolochic acid can promote the transdifferentiation of tubular epithelial cells.(2)Wenyanghuoxuefang have antagonistic role in renal tubular epithelial cells.