活性氧在高糖诱导人腹膜间皮细胞产生IL-8、MCP-1中的作用

目的 研究高糖对培养的人腹膜间皮细胞(human peritoneal mesothelial cells,HPMC)产生趋化蛋白IL-8(白介素-8)、MCP-1(单核细胞趋化蛋白-1)的影响以及内源性活性氧(过氧化氢)在其中的作用。方法 分别用不同浓度的葡萄糖、高渗甘露醇、葡萄糖加抗氧化剂(丙酮酸)刺激细胞,用半定量RT-PCR的方法测定趋化蛋白的基因水平,用ELISA的方法测定蛋白水平。结果 高糖可以使HPMC产生IL-8、MCP-1增加,且呈浓度依赖性,而高渗甘露醇的作用则不明显;高糖中加入抗氧化剂(丙酮酸)后,IL-8的表达下降,而MCP-1的表达未见明显的改变。结论 高糖可以使HPMC表达IL—8、MCP-1增加,从而在透析液生物不相容性及腹膜纤维化中起作用;在IL-8的产生中,内源性的活性氧可以作为信号传导分子,起到一定的作用。丙酮酸能够通过对抗内源性氧化应激,对HPMC具有保护作用。

Effect of high glucose on the expression of IL-8 and MCP-1 in human peritoneal mesothelial cells and the role of reactive oxygen species in this course

Objective To study the effects of glucose (0.1 % ,3.86 % ) on the production of IL - 8 and MCP - 1 in human peritoneal mesothelial cells ( HPMC) in vitro and the role of endogenous reactive oxygen species ( ROS) in this course.Methods The glucose (0.1%,3.86%) with or without hydrogen peroxide (0.1mmol/L,0.5rnmol/L) ,3.86% mannitoK glucose(3.86% ) with pyruvate (35mmol/L) were used in culture medium to stimulate the HPMC.IL- 8(In-terlukin- 8)mRNA and MCP - l(Monocyte chemoattractant protein - l)mRNA were measured by semiquantative RT -PCR,moreover,the protein level of IL - 8 by ELISA.Results The glucose enhanced the production of IL - 8 and MCP -1 following the increasing of concentration.On the contrary,the high osmolality mannitol had no these effects; Adding the antioxidant ( pyruvate) in the glucose(3.86% ) ,the expression of IL - 8 was lowered,but not for the MCP - 1.Conclusion The high glucose increases the expression of IL - 8 and MCP - 1,which may make a role in the uncomplicabality of dialysate and fibrosis of peritoneal membrane; The endogenous ROS can induce the expression of IL - 8 as a signal message .Pyruvate can protect the HPMC by counterworking the endogenous oxidant stress.