脂多糖作用下腹膜间皮细胞骨桥蛋白表达及乌司他丁的影响

The Expression of Osteopontin in Rat Peritoneal Mesothelial Cells Stimulated by Lipopolysaccharide and the Protective Effect of Ulinastatin

目的:观察脂多糖对大鼠腹膜间皮细胞(RPMC)中骨桥蛋白(OPN)表达的影响;观察乌司他丁(ulinastatin)作用下RPMC的OPN细胞分布及mRNA表达的影响。方法:胰蛋白酶法行RPMC的原代培养和传代,经鉴定第3代细胞融合至80%时分组。正常对照组:不同浓度LPS(50,100μg/ml)组;50μg/mlLPS作用不同时间(8,12,24,34,48h)组;乌司他丁组:(160,320,640U/ml)与LPS50mg/L作用24h,RealTime-RT-PCR技术检测骨桥蛋白(OPN)、TGF-β1mRNA的表达。结果:LPS可刺激RPMC的骨桥蛋白表达显著增加,呈剂量、时间依赖性(P<0.05);乌司他丁可降低LPS引起的RPMC中的骨桥蛋白表达,呈剂量依赖性(P<0.05)。结论:脂多糖通过上调骨桥蛋白的表达,引起腹膜损伤导致腹膜纤维化,乌司他丁可以一定程度抑制骨桥蛋白引起的腹膜纤维化过程。

Objective：To observe the expression of OPN in rat peritoneal mesothelial cells （RPMC） stimulated by Lipopolysaccharide. To observe the protective effect of Ulinastatin on rat peritoneal mesothelial cells （RPMC） fibrosis induced by lipopolysaccharide （LPS） and its protective mechanism in RPMC. Methods：RPMC were isolated from rat colic omentum and were incubated with LPS （ 50 μg/ml, 100 μg/ml） . The different dose（160,320,640 U/ml） of Ulinastatin was injected in the LPS（50μg/ml） group of RPMC.OPN was detected by Immunohistochemistry and RealTime-RT-PCR. TGF-β1 was detected by RealTime-RT-PCR.Results：Compared with the control group, the expression of OPN was significantly increased in the groups stimulated by LPS . In the group pre-incubated with LPS, compared with the group stimulated by LPS, the up-regulating expression of OPN was significantly inhibited. Ulinastatin decreased the expression of OPN and TGF-β1 which stimulated by Lipopoysaccharide（LPS）.Conclusion：LPS can up-regulate the expression of OPN in rat peritoneal mesothelial cell. Ulinastatin can down-regulate the fibrosis of RPMC stimulated by LPS through the down- regulation of OPN expression and activity.