固有免疫信号分子hsa-miR-146b调控脑卒中关联信号分子的生物信息学分析与实验研究

发布时间：2018-08-25 13:16

【摘要】：该研究旨在运用生物信息学方法预测固有免疫信号分子hsa-mi R-146b与脑卒中关联的分子调控网络。实验运用UCSC基因组浏览器、人类mi RNA疾病数据库、TF-mi RNA调控数据库、mi RNA靶基因预测验证数据库和Genecards数据库研究hsa-mi R-146b的上游转录因子、下游靶基因及信号通路的多个调控途径,绘制hsa-mi R-146b的核心调控网络图。采用脂多糖(lipopolysaccharide,LPS)刺激慢病毒感染的小胶质细胞BV2细胞,采用实时定量PCR检测早期生长反应基因1(early growth response 1,EGR1)、Toll样受体4(Toll-like receptor 4,TLR4)、mi R-146b、脑源性神经营养因子(brain derived neurotrophic factor,BDNF)和基质金属蛋白酶9(matrix metalloproteinase 9,MMP9)的m RNA水平变化,对hsa-mi R-146b调控网络进行验证。UCSC数据库中显示,hsa-mi R-146b在多个物种中具有高度保守性。生物信息学分析显示,hsa-mi R-146b受转录因子EGR1调控的同时,又调控下游TLR4、BDNF和MMP9等39个靶基因。所有基因构成一个以hsa-mi R-146b为核心的调控网络,在脑卒中的发生与发展中起着重要作用。在LPS刺激的BV2细胞中,EGR1、TLR4、mi R-146b、BDNF和MMP9m RNA水平升高,而RNA慢病毒技术干扰小胶质细胞中的EGR1的表达后,TLR4、mi R-146b和MMP9 m RNA水平降低,BDNF升高,表明EGR1是调节mi R-146b表达和下游信号分子TLR4、BDNF和MMP9的关键信号分子。综上所述,生物信息学方法预测并初步验证了hsa-mi R-146b分子在脑卒中疾病中的调控网络,为深入阐明hsa-mi R-146b在脑卒中疾病中的机制奠定了实验基础。
[Abstract]:The aim of this study was to predict the molecular regulatory network associated with stroke by bioinformatics. UCSC genome browser, TF-mi RNA regulatory database and Genecards database were used to study the upstream transcription factor, downstream target gene and signal pathway of hsa-mi R-146b. Draw the core control network diagram of hsa-mi R-146b. Lipopolysaccharide (lipopolysaccharide,LPS) was used to stimulate BV2 cells infected with lentivirus. Real time quantitative PCR was used to detect the changes of m RNA levels of Toll-like receptor 4 (Toll-like receptor 4 TLR4), brain-derived neurotrophic factor (brain derived neurotrophic factor,BDNF) and matrix metalloproteinase 9 (MMP9) in the early growth response gene 1 (early growth response 1 / EGR1). The verification of hsa-mi R-146b regulatory network shows that hsa-mi R-146b is highly conservative in many species. Bioinformatics analysis showed that hsa-mi R-146b was regulated by transcription factor EGR1, but also regulated 39 target genes such as downstream TLR4,BDNF and MMP9. All genes constitute a regulatory network with hsa-mi R-146b as the core, which plays an important role in the occurrence and development of stroke. In BV2 cells stimulated by LPS, the levels of BDNF and MMP9m RNA increased in BV2 cells stimulated by LPS, while RNA lentivirus technology inhibited the expression of EGR1 in microglial cells and decreased the levels of MMP9 m RNA and TLR4mi R-146b. It is suggested that EGR1 is the key signal molecule to regulate the expression of mi R-146b and the downstream signal molecules TLR4,BDNF and MMP9. In conclusion, the bioinformatics method predicts and preliminarily verifies the regulatory network of hsa-mi R-146b molecule in stroke disease, which lays an experimental foundation for further elucidating the mechanism of hsa-mi R-146b in stroke disease.
【作者单位】：南昌大学第一附属医院药学部;南昌大学转化医学研究院心血管疾病研究中心;
【分类号】：R743.3

【相似文献】