脂多糖对人脐静脉内皮细胞EPCR和NF-B表达的影响及参附注射液的干预作用  

Effects of Shenfu injection on expressions of EPCR mRNA, protein and NF-B protein in human umbilical vein endothelial cells induced by lipopolysaccharide

作　　者：朱金强 张近波 张小乐 许国斌 鄢来超 金晓红 

机构地区：[1]浙江省温岭市第一人民医院,浙江温岭317500 [2]河北省滦南县医院,河北滦南063500

出　　处：《现代实用医学》2010年第9期975-977,980,共4页Modern Practical Medicine

摘　　要：目的观察脂多糖(LPS)培养的人脐静脉内皮细胞内皮细胞蛋白C受体(EPCR)和核因子-B(NF-B)的表达,并探讨参附注射液对其的干预作用。方法分别在12、24及48h采用逆转录聚合酶链反应(RT-PCR)、蛋白免疫印迹技术测定正常对照组、LPS组、参附干预组培养的人脐静脉内皮细胞EPCRmRNA、蛋白及NF-B的表达。结果与正常对照组比较,LPS(24h)组EPCRmRNA、EPCR蛋白表达水平均低(均<0.01),LPS(48h)组EPCRmRNA、EPCR蛋白表达水平亦均低(均<0.05);与LPS(24h)组比较,正常对照组、参附(12h)组EPCRmRNA、EPCR蛋白表达水平均高(均<0.01),LPS(12h)组、参附(24、48h)组EPCRmRNA、EPCR蛋白表达水平亦高(均<0.05)。与正常对照组比较,LPS(24h)组NF-B蛋白表达水平显著增高(<0.01),LPS(48h)组NF-B蛋白表达水平亦高(<0.05);与LPS(24h)组比较,正常对照组、参附(12、48h)组NF-B蛋白表达水平均低(均<0.01),LPS(12h)组、参附(24h)组NF-B蛋白表达水平亦低(均<0.05)。结论 LPS可以从基因、蛋白水平减少人脐静脉内皮细胞上EPCR的表达,可能是通过NF-B途径实现的;参附注射液可以调节LPS对人脐静脉内皮细胞上EPCR的表达的影响,从而对脓毒症起到防治作用。Objective To investigate the effects of Shenfu injection on expressions of endothelial protein C receptor （EPCR ） mRNA, protein and NF-B in human umbilical vein endothelial cells （HUVECs） incubated with lipopolysaccharide（LPS）. Methods The EPCR mRNA, protein or phosphorylated protein of NF-B in HUVECs was detected by reverse transcription polymerase chain reaction （RT-PCR） or western bloting （WB） respectively. Results The expressions of EPCR mRNA and protein were significantly lower in LPS stimulation （24 h） （0.01） and in LPS stimulation （48 h） group （0.05） than those in normal control group. However, the expressions of EPCR mRNA and protein were significantly higher in Shenfu （12 or 24 h） groups than those in LPS stimulation （12 or 24 h ） group （0.01; 0.05）. The expressions of NF- B protein were significantly higher in LPS stimulation （24 and 48 h） groups than those in normal control group （0.05 ）; the expressions of NF-B protein were significantly lower in Shenfu treated （12, 24 h or 48 h） groups than that in LPS stimulation group （all 0.05）. Conclusion The data suggested that LPS could induce the decrease of EPCR mRNA and protein expressions through increasing the expression of NF- B in HUVECs, and Shenfu injection could block the inhibitory action of LPS on EPCR.

关 键 词：脂多糖类 人脐静脉内皮细胞 内皮细胞蛋白C受体 核因子-B 

分 类 号：R322.123[医药卫生—人体解剖和组织胚胎学] R363.21[医药卫生—基础医学]