蒙花苷对脂多糖诱导的血管内皮细胞炎症损伤的影响

Effect of buddleoside on lipopolysccharide-induced inflammatory injury of vascular endothelial cells

目的:在体外细胞水平上模拟血管内皮炎症损伤,探讨蒙花苷对血管内皮细胞炎症损伤的保护作用及其机制。方法:采用脂多糖(LPS)刺激人脐静脉内皮细胞(EA.hy926),加入不同浓度的蒙花苷处理,DAPI染色法测定EA.hy926与单核细胞(THP-1)的黏附能力,ELISA法检测细胞上清液中肿瘤坏死因子(TNF-α)和白介素-1(IL-1)水平,Western blot法检测细胞中p38、JNK、ERK1/2、p-p38、p-JNK、p-ERK1/2、核因子-κB(NF-κB)和细胞间黏附分子-1(ICAM-1)的蛋白表达。结果:蒙花苷(5、10、20mol/L)可显著抑制LPS诱导的EA.hy926与THP-1的细胞间黏附作用,减少EA.hy926分泌的炎性因子TNF-α和IL-1水平,降低细胞中JNK、ERK1/2、p-p38、p-JNK、p-ERK1/2、NF-κB和ICAM-1的蛋白表达。结论:蒙花苷可通过抑制NF-κB及其相关信号通路的活化来减少炎性黏附和炎性因子分泌,从而缓解LPS引起的血管内皮细胞炎症损伤。

Objective： To study the protective effect of buddleoside on lipopolysccharide-induced vascular endothelial cells inflammatory injury.Methods： The cytotoxicity of buddleoside in different concentrations was determined by MTT assay. DAPI staining was used to evaluate the effect of buddleoside on adhesion of EA. hy926 and THP-1. Many protein expression,such as p38,JNK,ERK1 /2,p-p38,p-JNK,p-ERK1 /2,NF-κB and ICAM-1,were detected by the Western blot analysis. ELISA method was used to measure the levels of TNF-α and IL-1. Results：Buddleoside could not influence the growth of EA. hy926 in 0. 1 ~ 20 mol / L. Buddleoside（ 5,10,20 mol / L） could inhibit the adhesion of EA hy926 and THP-1 stimulated by LPS. Western blot results indicated that buddleoside treatment leaded to a decrease in JNK,ERK1 /2,p-p38,p-JNK,p-ERK1 /2,NF-κB and ICAM-1. Furthermore,ELISA results showed that buddleoside reduced LPS-stimulated TNF-α and IL-1 production in EA. hy926. Conclusion： These data support that buddleoside can ameliorate LPS-induced inflammatory injury by inhibiting the activation of NF-κB signaling pathway,as well as decrease the inflammatory adhesion and inflammatory cytokine secretion.