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 D3的干预作用

摘要

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 -β mRNA 表达上调（P ﹤0．01）；与 ADR 组相比，四个药物干预组 TGF -β mRNA 表达明显下调（P ﹤0．01）。48 h 时与正常对照组相比，ADR 组 TGF -β mRNA 表达上调（ P ﹤0．01）；与 ADR 组相比，HVD 组与AHVD 组 TGF -β mRNA 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.