大鼠肾小管上皮细胞NRK-52E中VDR、PKC及其相互作用对Na DC1表达的影响

Effects of PKC,VDR and their interaction on expression of Na DC1 in rat renal tubular epithelial NRE-52E cells

目的:探究大鼠肾小管上皮细胞NRE-52E中蛋白激酶C(PKC)、维生素D受体(VDR)及其相互作用对钠离子依赖性二羧酸协同转运蛋白1(Na DC1)表达的影响。方法:体外培养大鼠肾小管上皮细胞NRE-52E,采用PKC激动剂和PKC抑制剂干预NRE-52E细胞,并用VDR过表达载体及VDR-shRNA载体转染NRE-52E细胞,结合Western blot检测细胞中PKC、VDR和Na DC1的蛋白表达情况。结果:成功建立VDR稳定过表达和VDR稳定干扰的NRK-52E细胞株。与对照组比较,PKC激动剂组VDR和Na DC1蛋白表达量显著上调(P <0. 01),VDR过表达组的PKC和Na DC1蛋白表达量显著上升(P <0. 01),VDR干扰+PKC激动剂组和VDR过表达+PKC抑制剂组中PKC和Na DC1蛋白的表达水平均介于VDR干扰组与VDR过表达组之间。结论:在大鼠肾小管上皮细胞NRE-52E中PKC活性增强诱导VDR蛋白表达,PKC活性减弱抑制Na DC1蛋白表达;VDR与PKC和Na DC1的蛋白表达存在明显正相关;PKC与VDR相互作用时PKC高活性和VDR过表达是促进或维持Na DC1蛋白表达的主要因素。

AIM:To investigate effects of protein kinase C(PKC),vitamin D receptor(VDR)and their interaction on expression of Na+/dicarboxylate contransporter 1(NaDC1)in rat renal tubular epithelial NRE-52E cells.METHODS:Rat renal tubular epithelial cell line NRE-52E was cultured in vitro.PKC agonist and inhibitor were used to interfere with NRE-52E cells.VDR over-expression and shRNA vectors were used to transfect into NRE-52E cells.The protein expression of PKC,VDR and NaDC1 in the cells was determined by Western blot.RESULTS:The NRK-52E cells with stable VDR over-expression and stable VDR interference were successfully set up.Compared with control group,the protein expression of VDR and NaDC1 in PKC agonist group and VDR over-expression group was increased significantly(P<0.01),and that in VDR interference combined with PKC agonist group and VDR over-expression combined with PKC inhibitor group was between VDR interference group and VDR over-expression group.CONCLUSION:In rat renal tubular epithelial cell line NRE-52E,enhanced PKC activity induces protein expression of VDR,while decrease in PKC activity inhibits the protein expression of NaDC1.There is a significantly positive correlation between VDR and regulation of PKC and NaDC1 protein expression.When PKC and VDR interact with each other,high activity of PKC and over-expression of VDR are the main factors to promote or maintain NaDC1 protein expression in the NRE-52E cells.