白鹤冲剂对TNF-α刺激的血管内皮细胞与嗜中性粒细胞黏附的影响

Influence of Baihe Decoxtion on the Polymorphonuclear-endotheliocyte Adhesion and the Activation of NF-kappaB

目的:观察白鹤冲剂对人脐静脉内皮细胞(HUVEC)与人外周血嗜中性粒细胞黏附以及细胞间黏附分子-1(ICAM-1)和CD18表达及核转录因子NF-κB(NF-κB)的影响,探讨白鹤冲剂治疗血管炎的作用机制。方法:将细胞分为正常对照组、TNF-α组、TNF-α+白鹤冲剂大剂量组、TNF-α+白鹤冲剂小剂量组和TNF-α+雷公藤内酯醇组,共五组。分别进行细胞黏附实验;以细胞ELISA法观察HVVEC的ICAM-1表达;以免疫细胞化学间接法检测CD18表达;间接免疫荧光法观察白鹤冲剂对HUVEC中NF-κB活化。结果:白鹤冲剂能抑制肿瘤坏死因子-α(TNF-α)刺激的HUVEC中ICAM-1表达和人外周血嗜中性粒细胞CD18表达(P<0.05),同时呈剂量依赖性抑制人外周血嗜中性粒细胞与HUVEC的黏附。结论:通过影响HUVEC中NF-κB活化从而抑制ICAM-1表达和嗜中性粒细胞CD18的表达,最终抑制嗜中性粒细胞与HUVEC的黏附可能是白鹤冲剂治疗血管炎的作用机制。

Objective： To study the effect of Baihe Decoxtion on the adhesion between human neutrophils and the umbilical vein endothelial cells （HUVEC） and the expression of intercellular adhesion molecule-1 （ICAM）-1 and CD18. Methods： The cells were divided into 5 groups： controlled group （Group A）, TNF-α group （GroupB）, TNF-α＋high dosage of Baihe Deeoxtion（GroupC）, TNF-α＋low dosage of Baihe Decoxtion（GroupD）, TNF-α＋Triptolide（Group E）. Cell adhesion tests were performed in each group. ELISA method was applied in testing the expression of ICAM-1 and indirect-immuno-cellchemistry method was applied in testing the expression of CD18. The activation of NF-κB in HUVEC affected by Baihe Decoxtion was observed through IIF. Results： Baihe Decoxtion depressed the expression of ICAM-1 in HUVEC stimulated by TNF-α and CD18 in polymorphonuelear and the activation of NF-κB. The depression was dosage dependent. Conclusion： It is an important mechanism in treating vasculitis with Baith Deeoxtion that it probably depresses the expression of ICAM-1 and CD18 in polymorphonuclear through effecting the activation of NF-κB in HUVEC, and finally depress the adhesion between polymorphonuelear and HUVEC.