论文LPS中华肾.doc

1,25(OH)2D3对脂多糖作用下大鼠腹膜间皮细胞维生素D受体及TNFα、TGF-β1表达的影响 杨丽娜，马健飞，陈硕，李丽燕，王力宁 [摘要]目的： 研究脂多糖(LPS)对大鼠腹膜间皮细胞(RPMC)维生素D受体(VDR)及 TNFα、TGF-β1表达的影响,从而为1,25(OH)2D3在腹透相关腹膜炎中的应用提供理论基础。方法：胰蛋白酶消化法原代培养腹膜间皮细胞、传代、经鉴定后分组：(1)正常对照组；(2)脂多糖组：不同浓度的脂多糖(1，10，100μg/ml)分别作用6h；10μg/ml脂多糖分别作用2，6，12h；(3) 1,25(OH)2D3作用组：10μg/ml脂多糖预孵育2h后，加1,25(OH)2D3(10-8mol/L,10-7mol/L,10-6mol/L)再作用6h。RT-PCR法检测VDR mRNA的表达；Western印迹法检测VDR蛋白表达；ELISA法检测上清液TNFα、TGF-β1的表达。结果：与对照组相比，LPS组RPMC VDR mRNA和蛋白表达下降(P 0.05)。与LPS组相比，1,25(OH)2D3组VDR mRNA和蛋白表达显著增强(P 0.01)。LPS组上清液中TNFα、TGF-β1浓度显著高于对照组 (P 0.01)；1,25(OH)2D3组上清液中TNFα、TGF-β1浓度显著低于LPS组 (P 0.01)。结论：LPS能下调RPMC VDR mRNA和蛋白的表达，上调TNFα、TGF-β1表达；1,25(OH)2D3可逆转LPS所致的RPMC VDR mRNA和蛋白的表达上调，并下调TNFα、TGF-β1表达。VDR与TNFα、TGF-β1表达负相关，VDR对腹膜透析相关腹膜炎具有一定的保护作用，并具有抑制腹膜纤维化的作用。 [关键词] 脂多糖；1,25(OH)2D3；维生素D受体；TNFα；TGF-β1；腹膜透析； Effects of 1,25(OH)2D3 on rat peritoneal mesothelial cells expression of vitamin D receptor, TNFα and TGF-β1 stimulated by lipopolysaccharide.YANG Li-na,MA Jian-fei,CHEN Shuo,LI Li-yan,WANG Li-ning. Department of Nephrology,The First Affiliated Hospital,China Medical University,Shenyang 110001,China Corresponding author: MA Jian-fei,Email:majiangfei@ [Abstract] Objective To observe the effect of 1,25(OH)2D3 on expression of vitamin D receptor(VDR),TNF-α(tumor necrosis factor-α)and TGF-β1 (transforming growth factor-β1) in rat peritoneal mesothelial cell(RPMC) stimulated by lipopolysaccharide(LPS),so as to providesome theoretic basis for the clinical use of 1,25(OH)2D3 on peritoneal dialysis associated peritoneal inflammation . Methods RPMCs were isolated,cultured and passaged by enzymatic disaggregation,then identified by phase contrast inverted microscope,transmission electron microscope with immunocytochemistry method.RPMCs were incubated with LPS(1，10，100μg/ml)and LPS(10μg/ml)for 2,6,12 hours,or stimulated by 1,25(OH)2D3(10-8mol/L,10-7mol/L,10-6mol/L)after incubated with LPS(10μg/ml)for 2 hours.RPMCs in the control group were just incubated with medium.VDR mRNA was d