盐酸戊乙奎醚抑制脂多糖诱导血管内皮细胞表达细胞间黏附分子-1及其机制

Penehyclidine hydrochloride inhibits lipopolysaccharide-induced intercellular adhesion molecule-1 expression in endothelial cells

目的 探讨盐酸戊乙奎醚（PHC）抑制脂多糖（LPS）诱导人脐静脉内皮细胞表达细胞间黏附分子（ICAM-1）及其作用机制.方法 体外培养人脐静脉内皮细胞株EVC-304,分别用LPS、PHC和LPS＋ PHC联合处理.Western blot及逆转录-聚合酶链反应（RT-PCR）检测ICAM-1的表达;Western blot检测丝裂原活化蛋白激酶（p38 MAPK）信号通路的激活；通过抑制剂SB203580阻断p38MAPK信号通路后,检测对ICAM-1表达的影响.结果 LPS可以诱导人脐静脉内皮细胞中磷酸化丝裂原活化蛋白激酶（p-p38 MAPK）表达增加113％ （P ＜0.05）,ICAM-1 mRNA表达增加105％,蛋白表达增加83％（P＜0.05）；联合应用PHC后,LPS诱导的p-p38MAPK表达增加下降36％（P＜0.05）;ICAM-1 mRNA表达增加下降40％,蛋白表达增加下降31％（P＜0.05）.通过SB203580阻断p38MAPK信号通路后,LPS诱导的ICAM-1 mRNA表达增加下降49％,蛋白表达增加下降39％（P＜0.05）.结论 LPS可以诱导血管内皮细胞表达黏附分子ICAM-1,PHC可以抑制这种LPS诱导的ICAM-1表达,PHC可能通过抑制p38MAPK信号通路的激活,从而抑制其下游细胞间黏附分子ICAM-1表达.

Objective To investigate the inhibitory effect of penehyclidine hydrochloride （PHC）on lipopolysaccharide （LPS）-induced expression of intercellular adhesion molecule-1 （ICAM-1） in human umbilical vein endothelial cells （hUVBECs） EVC-304.Methods EVC-304 cells were cultured and treated with PHC,LPS and PHC ＋ LPS （pretreated with PHC,followed by LPS） separately.The expression of ICAM-1 was detected by using reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting.The p38 mitogen-activated protein kinase （p38MAPK） expression and its phosphorylate-protein kinase （p-p38MAPK） were tested by using Western blotting.Finally,in another experiment,cells were pretreated with p38MAPK inhabitor SB203580,followed by LPS treatment,and the expression of ICAM-1 was examined by using RT-PCR and Western blotting.Results LPS added p-p38MAPK expression by 1.13 folds,and increased 1.05 folds of ICAM-1 mRNA and 0.83 folds of ICAM-1 protein in hUVBECs （P＜0.05）,which could be attenuated by the treatment of PHC （0.36 folds decrease of p-p38MAPK,0.4 folds of ICAM-1 mRNA and 0.31 folds of ICAM-1 protein,P ＜0.05）.The expression of ICAM-1 also could be restrainted by the treatment of p38MAPK inhabitor （0.49 folds decrease of mRNA and 0.39 folds of protein P ＜ 0.05）.Conclusion LPS could markedly increase the expression of ICAM-1,which could be inhibited by PHC.PHC may attenuate the expression of ICAM-1 by suppressing the activation of p38MAPK pathway,and PHC may have a protective effect against LPS-induced cell injury.