摘要
目的探讨糖皮质激素对人腹膜间皮细胞分泌细胞外基质的影响。方法采用胰蛋白酶消化法从人腹膜组织中分离间皮细胞,建立稳定的体外培养模型。将第三代细胞转板,至细胞融合后,将其分为:①对照组(F12);②2.5%葡萄糖组(F12+葡萄糖2.5%);③LPS组(F12+LPS10mg/L);④2.5%葡萄糖加DXM组(DXM为3种不同的浓度,依次为10nmol/L、100nmol/L、1000nmol/L);⑤LPS加DXM组(DXM为三种不同的浓度,依次为10nmol/L、100nmol/L、1000nmol/L);所有分组均培养24h后收集细胞上清液,低温离心(1000r/m)5min后,采用酶联免疫吸附法检测上清液中纤维连接蛋白(FN)和纤溶酶原抑制剂(PAI-1)的含量。结果①腹膜间皮细胞在2.5%葡萄糖和10mg/LLPS刺激下分泌FN及PAI-1明显增加,与对照组比较差异有显著性意义(P<0.01);10mg/LLPS组分泌FN及PAI-1较2.5%葡萄糖组分泌高,差异有显著性意义(P<0.01);②腹膜间皮细胞在2.5%葡萄糖加入不同浓度的DXM(10nmol/L、100nmol/L、1000nmol/L)干预后,24h内分泌FN及PAI-1较2.5%葡萄糖组减少,差异有显著性(P<0.05);③腹膜间皮细胞在LPS加入不同浓度的DXM(10nmol/L、100nmol/L、1000nmol/L)干预后,24h内分泌FN及PAI-1较LPS组减少,差异有显著性意义(P<0.05)。
Objective To study the effect of glucocorticoid on high concentration glucose and lipopolysaccharide（LPS） induced extracellular matrix secretion in cultured human peritoneal mesothelial cells. Methods Mesothelial cells were isolated from human omental specimens by trypsin disaggregation and cultured. Cultured human peritoneal mesothelial cells were treated with FI2 without serum （ Control group）, F12 with glucose （2.5% glucose group）, F12 with LPS（LPS group）, F12 with dexamethasone （DXM） 100 nmol/L （DXM group）,F12 with 2.5 %glucose and concentrations of DXM 10nmol/L-1000nmol/L（2.5% glucose and DXM group）, FI1 with LPS and concentrations of DXM 10 nmol/L-1000 nmol/L（ LPS and DXM group）. Enzyme-linked immunosorbent assay （ELISA） determined the quantity of FN and PAI-1 in culture supernatants. Results ① Compared with control group, the levels of FN and PAI-1 in culture supernatants with high concentration glucose and lipopolysaccharide were increased significantly （P〈0. 1）; ② The different concentration of dexamethasone decreased FN and PAl i secretion Compared with 2.5% glucose group at 24 hours（P〈0. 05）;③ The different concentration of dexamethasone decreased FN and PAI-I secretion compared with LPS. group at 24 hours（P〈0. 05）. Conclusion High concentration glucose and lipopolysaccharide can increase the levels of FN and PAI-1.Dexamethasone can inhibit the upregulation effect of high concentration glucose and lipopolysaccharide on FN and PAI-1 expression in cultured human peritoneal mesothelial cells.
出处
《中国血液净化》
2005年第7期375-378,共4页
Chinese Journal of Blood Purification
关键词
糖皮质激素
细胞外基质
腹膜间皮细胞
Glucocorticoid
Extracellular matrix
Peritoneal mesothelial cell
