文献类型：期刊文献

中文题名：活血定眩胶囊对小鼠脑微血管内皮细胞bEnd.3基因表达谱的影响及生物信息学分析

英文题名：Effect of Huoxue Dingxuan Capsule on gene expression profile in mice brain microvascular endothelial cell bEnd.3 and analysis in bioinformatics

作者：宋敏[1];刘涛[1];巩彦龙[1];周灵通[1];董万涛[2];刘建鸿[1]

第一作者：宋敏

机构：[1]甘肃中医药大学,甘肃兰州730000;[2]甘肃中医药大学附属医院,甘肃兰州730020

第一机构：甘肃中医药大学

年份：2017

卷号：48

期号：8

起止页码：1617

中文期刊名：中草药

外文期刊名：Chinese Traditional and Herbal Drugs

收录：CSTPCD;;Scopus;北大核心:【北大核心2014】；CSCD:【CSCD2017_2018】；

基金：国家自然科学基金资助项目(81360554);甘肃省中药现代制造工程研究院项目(YWW-2015049)

语种：中文

中文关键词：活血定眩胶囊;小鼠脑微血管内皮细胞;基因芯片;抗氧化;bEnd.3细胞

外文关键词：Huoxue Dingxuan Capsule; mouse brain microvascular endothelial cells; gene chip; anti-oxidant; bEnd.3 cells

摘要：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 Huoxue Dingxuan Capsule（HDC） containing serum on gene expression profile in mice brain microvascular endothelial cell bEnd.3 and its molecular mechanism. Methods Thirty rats were randomly divided into control group and HDC group, each group had 15 rats, and the serum was collected in two groups of rats. BEnd.3 was divided into blank serum group, hypoxia model group, and 10% HDC containing serum group. The cells were intervened in different conditions, and after oxygen deprivation for 6 h, the cytoskeleton was observed under laser microscope. And to detect the effect of HDC containing serum on gene expression profile in mice brain microvascular endothelial cell bEnd.3 by Affymetrix U133 plus2.0 gene expression microarray, and for clustering analysis by molecular annotation system MAS3.0. Results Differentially expressed genes were identified based on P〈0.01, ｜Fold Change｜〉3. There were 405 differentially expressed genes between blank group and model group, in which 176 genes were up-regulated and 229 genes were down-regulated. There were 368 differentially expressed genes between HDC sero group and model group, in which 146 genes were up-regulated and 222 genes were down-regulated. These differentially expressed genes had the functions of positive regulation of endothelial cell migration, process of acid metabolism, production of vascular endothelial growth factors, positive regulate activity of NF-κB transcription factors, oxidative stress-induced senescence, mitosis prophase, platelet aggregation（blockage formation）, ect. And many signaling pathways were regulated by these genes including vascular endothelial growth factor signaling pathway, interleukin signal pathway, p53 pathway, TNF-signal pathway, PPAR signaling pathway, PI3K-Akt signaling pathway, apoptosis signal pathway etc. Conclusion HDC has significant inhibitory effects on damage of bEnd.3 cells induced by hypoxia. Its mechanism is related to oxidative stress, inhibition of apoptosis, promotion of angiogenesis, inflammation, and immune response. Huoxue Dingxuan Capsule can regulate the function of vascular endothelial cell on gene level by several signaling pathways.