活性维生素D3对阿霉素诱导的足细胞转分化的干预作用∗

摘要

目的：：探讨活性维生素D3对阿霉素诱导的足细胞上皮-间充质细胞转分化( EMT)的干预作用。方法：以体外培养的小鼠足细胞系为研究对象,分六组：正常组( C组)、阿霉素组(即模型组,ADR组)、缬沙坦组( ARB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+高剂量活性维生素D3组( AHVD组)。干预24 h、48 h后采用RT-PCR检测synaptopodin、P-cadherin、desmin、FSP-1 mRNA的表达。 Western Bolt检测synaptopodin、P-cadherin、desmin蛋白的表达。结果：与C组相比,24 h及48 h ADR组synaptopodin、P-cadherin mRNA及蛋白表达均下降(P<0.01)、desmin mRNA及蛋白表达上调(P<0.01)和FSP-1 mRNA表达上调(P<0.01)。与ADR组相比,24 h时ARB组、LVD组、HVD组、AHVD组synaptopodin mRNA表达均升高(P<0.01)但四组之间差异无统计学意义。48 h时与ADR组相比,LVD组及HVD组synaptopodin mRNA及蛋白表达均升高(P<0.05)。48 h四个药物干预组P-cadherin mRNA及蛋白表达均升高(P<0.05)。24 h及48 h四个药物干预组desmin mRNA及蛋白表达均下调(P<0.01),FSP-1 mRNA表达均下调(P<0.05),但四组之间差异无统计学意义。结论：阿霉素可诱导小鼠足细胞发生EMT,活性维生素D3可通过修复足细胞synap-topodin、P-cadherin及抑制desmin、FSP-1的表达来拮抗足细胞EMT而起到保护足细胞的作用。缬沙坦和活性维生素D3在抑制足细胞EMT方面无协同作用。%Objective:To explore the effect of 1α, 25-dihydroxyvitamin D3 on αdriamycin-induced epithelial-mesen-chymal transition ( EMT) of podocytes. Methods:Mouse podocytes cultured in vitro under different differentiation conditions for 10 days were divided into 6 groups - the control group ( C group) , the experimental group ( ADR group) , the low dose 1α, 25-di-hydroxyvitamin D3 group ( LVD group) , the high dose 1α, 25-dihydroxyvitamin D3 group ( HVD group) , the valsartan group ( ARB group) and the combined group ( AHVD group:high dose 1α, 25-dihydroxyvitamin D3 combines and valsartan) . Each group was continuously intervened with drugs for 24 hours and 48 hours. The expression of protein molecules which were related to EMT such as synaptopodin, P-cadherin, desmin and FSP-1 were detected through reverse transcription polymerase chain reaction ( RT-PCR) . In addition, the expression of protein molecules such as synaptopodin, P-cadherin and desmin were also detected by Western Blot-ting. Results:Compared with the control group, after continuously intervened with drugs for 24 hours and 48 hours, in the ADR group, the expression of synaptopodin mRNA, protein, P-cadherin mRNA, and protein decreased, while the expression of desmin, mRNA, protein, and FSP-1 mRNA increased (P<0. 01). Compared with the ADR group, the expression of synaptopodin and mR-NA increased significantly in the ARB group, the LVD group, the HVD group and the AHVD group (P<0. 01). There was no signif-icant difference among the four drug intervention groups. In addition, the expression of synaptopodin, mRNA, and protein increased (P<0. 05) in the LVD group and the HVD group compared to the ADR group in 48 h. Compared with the ADR group, the expres-sion of P-cadherin and mRNA increased (P<0. 05) in the four drug invention groups. The expression of desmin mRNA and protein and FSP-1 mRNA significantly decreased (P<0. 01) in the four drug invention groups. Furthermore there was no significant differ-ence among the four groups. Conclusion:Adriamycin can induce podocytes injury through EMT. In vitro, 1α, 25-dihydroxyvitamin D3 can antagonize podocyte injury by repairing synaptopodin, P-cadherin expression while inhibiting desmin and FSP-1 expression. It shows that 1α, 25-dihydroxyvitamin D3 can protect podocytes through inhibiting EMT in podocytes. Valsartan and high dose 1α, 25-dihydroxyvitamin D3 have no synergy effect on the inhibition of podocyte EMT.