摘要
目的探讨七氟醚预处理抑制肿瘤坏死因子-α(TNF-α)诱导的血管内皮细胞细胞间黏附分子-1(ICAM-1)表达与c-Jun氨基末端激酶(JNK)的关系。方法采用随机数字表法将体外培养的人脐静脉内皮细胞随机分为5组,每组3孔:对照组(C组)、七氟醚预处理组(S组)、TNF-α处理组(T组)、七氟醚预处理加TNF-α处理组(S+T组)、JNK抑制剂SP600125加七氟醚预处理加TNF-α处理组(SP+S+T组)。S组、S+T组、SP+S+T组加入1.5MAC七氟醚饱和缓冲液,每次孵育15 min,共3次,间隔15 min。SP+S+T组在七氟醚预处理前1 h每孔加入30μmol SP600125。七氟醚预处理结束40 min后,T组、S+T组、SP+S+T组人脐静脉内皮细胞培养基中加入10 ng/ml的TNF-α建立炎症模型。应用Western blot法测定各组细胞JNK磷酸化水平(p-JNK)及ICAM-1的表达量。结果与C组相比,S组人脐静脉内皮细胞ICAM-1、p-JNK表达无明显差异(P>0.05),T组、S+T组ICAM-1、pJNK表达增高(P<0.05);与T组比较,S+T组ICAM-1、p-JNK表达下降(P<0.05);与S+T组比较,SP+S+T组ICAM-1、p-JNK表达降低(P<0.05)。结论七氟醚预处理抑制TNF-α诱导的血管内皮细胞ICAM-1表达与降低p-JNK有关。
Objective To investigate the role of c-Jun N-terminal kinase （JNK） in sevoflurane preconditioning inhibits tumor necrosis factor-α （TNF-α） induced intercellular adhesion molecule-1 （ICAM-1） expression in vascular endothe-lial cells. Methods Human umbilical vein endothelial cells were randomly divided into 5 groups with 3 wells in each group：normal control group （group C）,TNF-α treatment group （group T）,sevoflurane preconditioning group （group S）, sevoflurane preconditioning and TNF-α treatment group （group S＋T）,JNK inhibitor SP600125 and sevoflurane precon-ditioning and TNF-α treatment group （group SP＋S＋T）.1.5MAC sevoflurane saturated buffer was added to the culture medium and the cells underwent 3 episodes of 15 min incubation with sevoflurane at 15 min interval in group S,group S＋T and group SP＋S＋T.The cells were incubated with SP600125 （30 μmol） 1 h before sevoflurane preconditioning in group SP＋S＋T.Human umbilical vein endothelial cells were treated with TNF-α （10 ng/ml） in group T,group S＋T and group SP＋S＋T 40 min after the end of sevoflurane preconditioning.The expression level of ICAM-1 protein and the phosphorylation level of JNK were measured by Western blot. Results Compared with group C,the expression level of ICAM-1 and the phosphorylation level of JNK were activated by TNF-α in group T and group S＋T （P〈0.05）,but was not significant in group S （P〉0.05）.Compared with group T,sevoflurane pretreatment could decrease the expression level of ICAM-1 and p-JNK in group S＋T （P〈0.05）.Compared with group S＋T,ICAM-1 and p-JNK expression was down-regulated in group SP＋S＋T （P〈0.05）. Conclusion The reduction of p-JNK expression plays a role in sevoflurane pre-conditioning inhibits TNF-αinduced ICAM-1 expression in vascular endothelial cells.
出处
《中国当代医药》
2015年第7期4-7,共4页
China Modern Medicine
基金
国家自然科学基金资助项目(81270185)
