Effect Of Norcantharidin On Wnt/β-catenin In Obstructive Nephropathy

ObjectiveEstablish unilateral ureter obstructive (UUO) rats model and human proximal renal tubular epithelial cell(HK-2) fibrosis model in order to study two problems:1To detect the expression of β-catenin in UUO rats kidney after different obstructive time periods,and define the relationship between β-catenin and renal obstruction.2In different concentration of NCTD interference with UUO rats and HK-2cell stimulated by TGF-β,is to discuss the effection of NCTD on β-catenin, Wnt4.Methods1.animal experiments:We had two rats experiments,16Sprague-Dawley rats were randomly divided into control, UUO day3, UUO day7and UUO day14groups. Another16rats were randomly divided into control, UUO day14,NCTD (0.05mg/kg) interference group、NCTD0.1mg/kg interference groups.2. cell experiments:set control group, TGF-β15ng/ml stimulated group, TGF-β15ng/ml stimulated+NCTD2.5mg/L interference group,TGF-β15ng/ml stimulated+NCTD5mg/L interference group3. to detect protein expression of β-catenin,Wnt4by immunohistochemistry and western blot.4. to detect mRNA expression of β-catenin, Wnt4by RT-PCR.Results1.With prolonging ureter obstruction time, protein expression of β-catenin was increasing gradually, it reached the highest level in UUO day14group.2.Compared to the UUO group, in NTCD group protein and mRNA expressions of β-catenin,Wnt4decreased,the difference is statically significant.3.The β-catenin、wnt4protein expression in the TGF-β1stimulating HK-2group was up-regulated compared with controll group; The β-catenin、wnt4protein expressions in different concerntrations of NTCD interfereing group were down-regulated,and in concerntration dependant manner, the difference is statically significant.Conclusions1The β-catenin、wnt4expression in the obstructive renal tissue and HK-2cell stimulated by TGF-β1were increased.2NTCD interference can down-regulate the β-catenin、wnt4expression in the obstructive renal tissue and HK-2cell stimulated by TGF-β1.