阿霉素诱导的足细胞转分化中β-catenin及TGF-β的变化及活性维生素D_3的干预作用

The Role of β-catenin、TGF-β in Adriamycin-induced Podocytes Transdifferentiation and the Effect of 1α,25-dihydroxyvitamin D_3

目的:探讨阿霉素诱导的足细胞转分化(EMT)中β-catenin及TGF-β的变化及活性维生素D3干预作用。方法:以体外培养的小鼠足细胞系为研究对象,分6组进行实验:正常组(C组)、阿霉素组(即模型组,ADR组)、低剂量活性维生素D3组(LVD组)、高剂量活性维生素D3组(HVD组)、缬沙坦组(ARB组)、缬沙坦+高剂量活性维生素D3组(AHVD组)。各实验组干预24 h、48 h时通过RT-PCR检测β-catenin、TGF-βmRNA的表达,Western Bolt检测β-catenin蛋白的表达。结果:用药物干预24 h及48 h时,与正常对照组相比,ADR组足细胞β-catenin mRNA及蛋白表达均上调(P<0.01);与ADR组相比,四个药物干预组(ARB组、LVD组、HVD组、AHVD组)β-catenin mRNA及蛋白表达均下调(P<0.05)。24 h时与正常对照组相比,ADR组TGF-βmRNA表达上调(P<0.01);与ADR组相比,四个药物干预组TGF-βmRNA表达明显下调(P<0.01)。48 h时与正常对照组相比,ADR组TGF-βmRNA表达上调(P<0.01);与ADR组相比,HVD组与AHVD组TGF-βmRNA表达明显下调(P<0.01),而LVD组及ARB组无显著下降(P>0.05)。结论:活性维生素D3通过抑制信号转导通路中β-catenin、TGF-β的表达而抑制足细胞发生EMT。

Objective：To explore the role of β - catenin、TGF - β in adriamycin - induced epithelial - mesenchymal transi-tion（EMT）of podocytes and the effect of 1α,25 - dihydroxyvitamin D3 on it. Methods：Mouse podocytes in vitro cultured under dif-ferentiating conditions for 10 days were divided into 6 groups which were the control group（C group）,the model group（ADR group）, the low dose 1α,25 - dihydroxyvitamin D3 group（LVD group）,the high dose 1α,25 - dihydroxyvitamin D3 group（HVD group）,the valsartan group（ARB group）and the combination group（AHVD group,high dose 1α,25 - dihydroxyvitamin D3 combines with valsar-tan）. Each group was continually incubated with drugs for 24 h and 48 h. The expression of β - catenin and TGF - β mRNA which were related to EMT was detected by reverse transcription polymerase chain reaction（RT - PCR）. In addition,the expression of β -catenin protein was also detected by Western Blot. Results：After continually incubated with drugs for 24 h and 48 h,the expression of β - catenin mRNA and protein were increased in ADR group（P ﹤ 0. 01）;then compared with ADR group,the expression of β -catenin mRNA and protein decreased（P ﹤ 0. 05）in ARB group,LVD group,HVD group and AHVD group. In our study,the expres-sion of TGF - βmRNA increased（P ﹤ 0. 01）in ADR group when compared with control group in 24 h;then compared with ADR group,the expression of TGF - β mRNA decreased significantly（P ﹤ 0. 01）in the four drug intervention groups. Moreover,it showed that the expression of TGF - β mRNA increased（P ﹤ 0. 01）in ADR group when compared with control group in 48 h;then compared with ADR group,the expression of TGF - β mRNA decreased significantly（P ﹤ 0. 01）in HVD group and AHVD group,but ARB group and LVD group had no significant decreased（P ﹥ 0. 05）. Conclusion：1α,25 - dihydroxyvitamin D3 suppresses podocytes oc-curring EMT by inhibiting the expression of β - catenin and TGF - β which are signal transduction factors of podocytes EMT signal transduction pathway.