活血及活血解毒配伍中药含药血清对肿瘤坏死因子-α诱导人内皮细胞与中性粒细胞黏附及相关通路蛋白表达的影响  

Effect of Chinese Herbal Drug-Containing Serum for Activating Blood, Activating Blood and Dispelling Toxin on TNF-α-Induced Adherence between Endothelial Cells and Neutrophils and the Expression of MAPK Pathway

作　　者：郭春雨[1] 马晓娟[2] 刘倩[2] 殷惠军[3] 史大卓[2] 

机构地区：[1]首都医科大学附属北京妇产医院中医科,北京100026 [2]中国中医科学院西苑医院心血管中心,北京100091 [3]甘肃中医学院附属医院心血管科,兰州730000

出　　处：《中国中西医结合杂志》2015年第2期204-209,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然基金资助项目(No.81030063;No.81102845)

摘　　要：目的探讨活血及活血解毒配伍中药含药血清对肿瘤坏死因子-α(TNF-α)诱导人脐静脉内皮细胞(HUVECs)与外周血中性粒细胞(PMN)黏附、炎症反应及相关通路蛋白的影响。方法采用随机数字表法将32只大鼠分为空白对照组、芎芍胶囊组、黄连胶囊组及芎芍胶囊加黄连胶囊组,每组8只,各给药组按临床等效剂量灌胃,采血制备含药血清。空白对照组予等体积蒸馏水作对照。常规方法培养HUVECs,以TNF-α(100 ng/m L)为刺激物,制备HUVECs细胞损伤模型。细胞分为5组:空白对照组、模型组、活血组、解毒组及活血解毒组。空白对照组及模型组给予正常大鼠血清,其他3组以10%含药血清干预24 h后收集细胞,应用孟加拉玫瑰红染色法观察HUVECs与PMN发生病理性黏附情况;酶联免疫法检测细胞上清液相关炎症因子E选择素(E-selectin)、细胞黏附分子-1(ICAM-1)及白细胞介素-1β(IL-1β)含量;Western blot法检测丝裂素活化蛋白激酶p38(p38MAPK)及细胞外信号调节激酶1/2(ERK 1/2)蛋白磷酸化水平。结果与空白对照组比较,模型组细胞出现明显的氧化损伤,PMN黏附数量明显增加,IL-1β、E-selectin及ICAM-1水平升高,且HUVEC上清p-p38MAPK、p-ERK 1/2蛋白表达增加(均P<0.01)。与模型组比较,活血组、解毒组及活血解毒组HUVECs与PMN黏附数量减少(P<0.05),细胞培养上清液IL-1β、E-selectin及ICAM-1水平降低(P<0.05,P<0.01),且内皮细胞p-p38MAPK、p-ERK 1/2蛋白表达降低(P<0.01)。结论活血及活血解毒含药血清能减轻TNF-α诱导的HUVECs损伤,抑制HUVECs-PMN黏附和黏附因子释放。其机制可能与调节内皮细胞MAPK通路p38MAPK、ERK 1/2蛋白磷酸化水平有关。Objective To explore the effect of drug-containing serum of Chinese herbal com- pounds [Xiongshao Capsule （XS, for activating blood） and Huanglian Capsule （HL, for dispelling toxin）l on tumor necrosis factor-a （TNF-a）-induced adherence between human umbilical vein endothelial cells （HUVECs） and polymorphonuclear neutrophils （PMN）, inflammatory reaction and expression of related proteins in mitoqen-activated protein kinase （MAPK） pathway. Methods Thirty-two rats were randomly di-vided into four groups （8 in each group） using random digit table： the blank control group treated with distilled water, the test group I treated with Chinese herbal compound of XS （0. 135 g/kg）, the test group H treated with Chinese herbal compound of HL （0. 135 g/kg）, and the test group m treated with Chinese herb- al compound of XS （0. 135 g/kg） and HL （0. 135 g/kg）. All medication was given by gastrogavage once a day for a week. Rats＇ blood serum was harvested 1 h after the last administration to prepare drug-containing serum. HUVECs were exposed to TNF-eL （100 ng/mL） to induce cell injury model and incubated with corresponding drug-containing serum （10%） for 24 h. Normal rats＇ serum was given to cells in the blank control group and the model group, while XC ＋ HL containing serum was given to cells in the rest 3 groups. The adherence of HUVECs and PMN cells was detected by using rose bengal strain. Levels of E-selectin, intercellular adhesion molecule-1 （ICAM-1）, and interleukin-1β（IL-1β） in the supernatant of cultured HU- VECs were determined by ELISA. Protein expressions of mitogen-activated protein kinases p38 （p38MAPK） and extracellular signal-regulated kinase 1/2 （ERK 1/2）were determined by Western blot. Results Compared with the blank control group, HUVECs were seriously injured; PMN adherence amount significantly increased; levels of E-selectin, ICAM-1, and IL-1 β increased; expression levels of p-p38MAPK and p-ERK 1/2 in the supernatant of HUVECs sig

关 键 词：活血解毒 内皮细胞 细胞黏附 丝裂素活化蛋白激酶p38 细胞外信号调节激酶1/2 

分 类 号：R285[医药卫生—中药学]