白细胞介素-10对脂多糖直接诱导的血管内皮细胞损伤的影响

目的 :观察脂多糖 (LPS)对体外培养的人脐静脉内皮细胞 (HU VEC)凋亡和坏死的直接作用以及IL 10对其影响。方法 :将体外培养的HUVEC随机分为正常对照组、LPS组、观察组 (LPS +IL 10 ) ,各组在相应的培养液中于 3 7℃、5 %的CO2 孵育箱中培养。四唑盐(MTT)比色法检测不同时间细胞增殖的变化 ;酶联免疫吸附试验(ELISA)法检测各组vWF水平 ;用Annexin v及PI双标染色 ,流式细胞仪测定HUVEC的凋亡和坏死的变化。结果 :MTT结果示不同浓度LPS在 2 4h后对细胞增殖有抑制作用 (P <0 .0 5 ) ,并随培养时间延长和浓度增大 ,其抑制率逐渐增大 ,IL 10对细胞抑制率无明显改善 (P>0 .0 5 ) ;培养 2 4h后LPS组和观察组vWF水平明显升高 ,与对照组有显著性差异 ;流式细胞仪检测示LPS组及观察组细胞凋亡及坏死增高 ,以坏死细胞为主 ,与对照组均有显著性差异 (P <0 .0 1) ,而LPS组与观察组无显著性差异 (P >0 .0 5 )。结论 :LPS可以直接诱导血管内皮细胞损伤 ,而IL 10对其无明显的影响

Effect of IL-10 on the Direct Injury Induced by Lipopolysaccharide in Cultured Human Umbilical Vascular Endothelial Cells

Objective: To observe the direct injury of lipopolysaccharide (LPS) on cultured human umbilical vascular endothelial cells (HUVEC) and the effect of IL 10 on it.Method: The cultured HUVEC in vitro were randomly divided into control group, LPS group and experimental group (LPS+IL 10) and incubated at 37℃ in 5% CO 2 in incubator. Cellproliferation was assessed using MTT assay in different culture time; vWF levels were measured by ELISA method; Cell apoptosis and necrosis were detected by flow cytometry (FCM) using Annexin v PI staining.Results: MTT assay showed that cellproliferation are inhibited in LPS groups and experimental groups after 24 hours culture and the inhibitory rates are time and dose dependent,IL 10 haven′t improved effect on it( P >0.05); The levels of vWF increased highly in LPS groups and experimental groups after 24 hours culture, which is significantly different when compared with control groups. Flow cytometry (FCM) analysis indicated significant apoptosis and necrosis are found in all groups except controls, cell death occurred mainly in the form of necrosis and there is no significant difference between LPS group and experimental group( P >0.05).Conclusion: LPS may induce the direct injury of HUVEC including apoptosis and necrosis,while IL 10 have no apparent effects on it.