【摘要】 目的探讨p38有丝分裂素激活蛋白激酶(MAPK)对高糖诱导的人腹膜间皮细胞(HPMC)p27kip1的表达和纤连蛋白(FN)分泌的影响。方法同步化生长融合的HPMC在有或无p38MAPK抑制剂SB203580的条件下和不同浓度的葡萄糖共同孵育。采用Bradford法测定细胞内总蛋白量。采用逆转录-聚合酶链反应(RT-PCR)方法检测p27kip1mRNA的表达。p27kip1和p38MAPK蛋白用Western印迹法测定。采用酶联免疫吸附试验(ELISA)测定细胞培养液FN水平。结果高糖刺激HPMC时细胞内总蛋白量、p27kip1蛋白及细胞培养液里的FN蛋白水平明显升高。抑制p38MAPK活性可有效阻断高糖介导的腹膜间皮细胞p27kip1的表达及FN的分泌。结论高糖通过p38MAPK的活化可上调HPMCp27kip1的表达和FN的分泌。更多还原

【Abstract】 Objective To investigate the effect of p38 MAPK on the p27kip1 expression and fibronectin(FN)secretion in cultured human peritoneal mesothelial cells (HPMCs) under high glucose (HG) conditions. Methods HPMCs were isolated from human omentum and subcultured. After serum restriction, HPMCs were exposed to 5 6 mmol/L glucose (NG), 5 6 mmol/L glucose+44 6 mmol/L mannitol(NG+MN),or 50 mmol/L glucose (HG) for 48 to 72 hours with or without p38 MAPK inhibitor SB203580. RT PCR, Western blot and ELISA were performed to determine mRNA, protein and FN expression, respectively. Results p27kip1 protein expression and total protein content in HPMCs cultured under HG conditions significantly increased at 48 hours and 72 hours. HG induced fibronectin production by HPMCs was also significantly increased at 72 hours. HPMCs exposed to HG exhibited significant increases in p38 MAPK activity at 48 hours and remained higher levels until 72 hours. Treatment with SB203580 significantly inhibited HG induced p27kip1 protein levels and total protein content in HPMCs. HG induced fibronectin production by HPMCs was also significantly reduced by SB203580.Conclusion p27kip1 and FN increase in HPMCs exposed to HG via p38 MAPK activation, and activated p38 MAPK pathway may play an important role in the pathogenesis of peritoneal fibrosis.更多还原