PPARγ配体对高糖诱导大鼠腹膜间皮细胞结缔组织生长因子和纤溶酶原激活抑制因子1表达的影响  

Influence of peroxisome proliferators-activated receptor γ ligands on the expression of connective tissue growth factor and plasminogen activator inhibitor 1 in rat peritoneal mesothelial cells induced by high glucose

作　　者：周光宇[1] 马健飞[2] 李德天[1] 李丽燕[2] 李志明[2] 王力宁[2] 

机构地区：[1]中国医科大学附属盛京医院肾内科,沈阳110001 [2]中国医科大学附属第一医院肾内科

出　　处：《中华肾脏病杂志》2010年第2期123-127,共5页Chinese Journal of Nephrology

摘　　要：目的研究过氧化物酶体增殖物活化受体1（PPA脚）天然配体15d—PGJ2及人工合成配体吡格列酮（pioglitazone）对高糖诱导大鼠腹膜间皮细胞（RPMC）表达结缔组织生长因子（CTGF）和纤溶酶原激活抑制因子1（PAI-）的影响。方法胰蛋白酶消化法分离培养RPMC，经鉴定分组：（1）0．1％、1．5％、2．5％、4．25％葡萄糖作用24h组；（2）2．5％葡萄糖作用0、6、12、24、36、48、72h组；（3）0．1％、1．5％、2．5％、4．25％甘露醇作用24h组；（4）15d—PGJ2（5、15μmol/L）及吡格列酮（5、15μmol／L）分别预孵育2h，加2．5％葡萄糖再作用24h。RT—PCR检测CTGF和PAI1 1mRNA表达；Western印迹检测PPARγ、CTGF及PAI-1蛋白表达。结果正常RPMC有PPARy表达。1．5％葡萄糖使RPMC的PPARy蛋白表达减少（P〈0．05），而4．25％葡萄糖作用最大（P〈0．01）；2．5％葡萄糖作用6h，RPMC的PPARγ／蛋白表达减少（P〈0．05），m72h达高峰（P〈0．01）。各种浓度的甘露醇作用24h，RPMC的PPARγ蛋白表达均无明显变化（P〉0．05）。2．5％葡萄糖作用后RPMC的CTGF和PAI-1mRNA和蛋白表达均显著增加（P〈0．01）。5μmol／L的毗格列酮显著降低CTGF和PA1-1mRNA和蛋白表达（均P〈0．05），而15μmol／L作用更强（P〈0．01）。5μmol／L的15d—PGJ2显著降低RPMC的CTGFmRNA和蛋白表达以及PA1-1mRNA的表达（均P〈0．05），但不影响PA1-1蛋白表达（P〉0．05）．15μmol／L的15d—PGJ2对CTGF和PA1-1mRNA和蛋白表达均有抑制作用（P〈0．05或P〈0．01）。结论葡萄糖以时间和剂量依赖方式调节RPMCPPAR叫的表达，其作用与高渗透浓度无关。PPARγ配体可显著抑制高糖诱导的CTGF和PAI-1的表达，提示激活PPARγ可能成为防治腹膜透析相关腹膜纤维化的新途径之一。Objective To investigate the intluence of peroxisome proliferators-activated receptor γ （PPARγ） ligands on the expression of connective tissue growth factor （CTGF） and plasminogen activator inhibitor 1 （PAl-I） in rat peritoneal mesothelial cells （RPMCs） induced by high glueose. Methods RPMCs were isolated, cultured, passaged, confirmed and divided into 4 groups： （1）treated with glucose for 24 h in the concentrations of 0.1%, 1.5%, 2.5% and 4.25%, respectively; （2）treated with 2.5% glucose for 0, 6, 12, 24, 36, 48 and 72 h respectively; （3） treated with mannitol for 24 h in the coneentrations of 0.1%, 1.5%, 2.5% and 4.25%, respectively; （4）pretreated with pioglitazone （5, 15 μmol/L） and 15d-PGJ2 （5, 15 μmol/L） tbr 2 h respectively, then treated with 2.5% glucose for 24 h. The expression of CTGF and PAI-hnRNA was detected by RT-PCR. The expression of PPARγ, CTGF and PAI-1 protein was detected by Western blot. Results RPMCs expressed PPAR＇y in normal condition. Glucose reduced the protein expression of PPARγ in doseand time-dependent manner （P〈0.05 and P〈0.01）. Mannitol of different concentrations had no efl）et on the expression of PPARγ in RPMCs （all P〉0.05）. The mRNA and protein expression of CTGF was up-regulated significantly by stimulation of 2.5% glucose （P〈0.01）. Both 15d-PGJ2 and pioglitazone （5, 15 μmol/L） decreased the mRNA and protein expression of CTGF in concentration-dependent manner （P〈0.05 and P〈0.01）. 2.5% glucose increased the mRNA and protein expression of PAI-1 （P〈0.01）. 5 μmol/L 15d-PGJ2 decreased the mRNA expression of PAI-1, but not protein. 15 μmol/L 15d-PGJ2 decreased the mRNA and protein expression of PAI-1 （P〈0.05 or P〈0.01）. The mRNA and protein expression of PAI-1 alos down-regulated in concentration-dependent manner induced by pioglitazone （5, 15 μmol/L） （P〈0.05 and P〈0.01）. Conclusions High glucose down-regulates the expression of PPARγ in RPMCs and this effect is not con

关 键 词：过氧化物酶体类 腹膜透析 结缔组织生长因子 纤溶酶原激活抑制因子1 

分 类 号：R285.5[医药卫生—中药学]