目的探讨活血定眩胶囊含药血清对体外培养小鼠脑微血管内皮细胞bEnd.3基因表达谱的影响及作用的分子机制。方法将30只大鼠随机分为对照组和活血定眩胶囊组,2组分别ig给予7.5 g/kg活血定眩胶囊和等量生理盐水,连续7 d,采集2组大鼠血清。将bEnd.3细胞分为对照组、模型组、10%活血定眩胶囊含药血清组,给药后,bEnd.3细胞缺氧6 h。激光显微镜下观察细胞骨架,运用Affymetrix U133 plus2.0基因组表达芯片检测活血定眩胶囊对bEnd.3细胞基因表达谱的影响,应用分子注释系统MAS3.0软件进行聚类分析。结果根据P<0.01,|Fold Change|>3筛选差异基因,对照组与模型组之间共有405个差异基因,其中表达上调的有176个,表达下调的有229个。活血定眩胶囊含药血清组与模型组之间共有368个差异基因,其中表达上调的有146个,表达下调的有222个。这些共同的差异基因生物功能包括内皮细胞迁移的正调控、含氧酸代谢过程、血管内皮生长因子的产生、核转录因子-κB(NF-κB)活性的正向调节等,参与的信号通路包括氧化应激诱导的衰老、有丝分裂前期、血小板聚集(堵塞形成)、血管内皮生长因子信号通路、白细胞介素信号通路、p53通路、肿瘤坏死因子-α(TNF-α)信号通路、过氧化物酶体增殖物激活受体(PPAR)信号通路、磷脂酰肌醇3激酶-蛋白激酶B(PI3K-Akt)信号通路、凋亡信号通路等信号通路。结论活血定眩胶囊对缺氧所致bEnd.3细胞的损伤具有明显的对抗作用,其机制与氧化应激、抑制细胞凋亡、促进血管内皮新生、炎症及免疫反应有关,活血定眩胶囊在基因水平通过多条通路调节血管内皮细胞功能。 Objective To investigate the effect of Huoxingdingsuo capsule containing serum on the expression of bEnd.3 gene in mouse brain microvascular endothelial cells in vitro and its molecular mechanism. Methods Thirty rats were randomly divided into control group and Huoxundingngxuan Capsule group. Two groups were given 7.5g / kg Huoxuoding capsule and an equal volume of normal saline for two consecutive days. Two groups of rat serum were collected. BEnd.3 cells were divided into control group, model group and 10% Huoxuedinghuang capsule containing drug serum group. After administration, bEnd.3 cells were exposed to hypoxia for 6 hours. The cytoskeleton was observed under a laser microscope. Affymetrix U133 plus 2.0 genomic expression chip was used to detect the effect of Huoxuedingzhu capsule on the gene expression profile of bEnd.3 cells. Cluster analysis was performed using molecular annotation system MAS3.0 software. Results According to P <0.01, | Fold Change |> 3, a total of 405 differential genes were screened out from the control group and the model group, among which 176 were up-regulated and 229 were down-regulated. There are 368 differential genes between Huoxuedinghuang capsule containing drug serum group and model group, of which 146 genes are up-regulated and 222 genes are down-regulated. These common biological functions of differential genes include the positive regulation of endothelial cell migration, the oxoacid metabolism, the production of vascular endothelial growth factor, and the upregulation of nuclear transcription factor-κB (NF-κB) activity. The involved signaling pathways include Oxidative stress-induced senescence, pre-mitosis, platelet aggregation (blockage), vascular endothelial growth factor signaling pathway, interleukin signaling pathway, p53 pathway, tumor necrosis factor-alpha (TNF- alpha) signaling pathway, peroxidase (PPAR) signal pathway, PI3K-Akt signaling pathway and apoptosis signaling pathway. Conclusion Huoxueding Capsule has a significant antagonistic effect on the injury of bEnd.3 cells induced by hypoxia. The mechanism is related to oxidative stress, inhibition of apoptosis, promotion of endothelium neovascularization, inflammation and immune response. Genes regulate vascular endothelial function through multiple pathways.