急性缺氧致小鼠脑血管平滑肌功能改变的转录组学分析  

Transcriptome analysis of acute hypoxia-induced functional changes in cerebrovascular smooth muscle in mice

作　　者：常玮 李亚娟 臧克海 李少华 刘美杰 刘峰舟 薛军辉 CHANG Wei;LI Yajuan;ZANG Kehai;LI Shaohua;LIU Meijie;LIU Fengzhou;XUE Junhui(Center of Clinical Aerospace Medicine,School of Aerospace Medicine,Xijing Hospital,Air Force Medical University,Xi'an 710032,China;Department of Aerospace Physiology,School of Aerospace Medicine,Xijing Hospital,Air Force Medical University,Xi'an 710032,China;Department of Recuperation and Rehabilitation for Flight Personnel,School of Aerospace Medicine,Xijing Hospital,Air Force Medical University,Xi'an 710032,China;Department of Aviation Medicine,Xijing Hospital,Air Force Medical University,Xi'an 710032,China)

机构地区：[1]空军军医大学航空航天医学系航空航天临床医学中心,陕西西安710032 [2]空军军医大学航空航天医学系航空航天生理学教研室,陕西西安710032 [3]空军军医大学航空航天医学系飞行人员疗养与康复教研室,陕西西安710032 [4]空军军医大学西京医院空勤科,陕西西安710032

出　　处：《空军军医大学学报》2024年第2期198-205,共8页Journal of Air Force Medical University

基　　金：陕西省重点研发计划项目(2022SF-246);军队实验动物专项青年项目(SYDW[2020]23)。

摘　　要：目的探讨小鼠脑血管平滑肌细胞(CVSMCs)在低气压缺氧脑损伤中的作用及其可能的发生机制。方法将雄性C57BL/6J小鼠随机分为平原组(Sham组)和高原组(HA组),每组10只。采用HE染色、免疫组织化学法、免疫荧光法、Western blotting观察各组小鼠脑损伤以及脑血管平滑肌功能变化情况。分离培养另20只C57BL/6J小鼠的原代CVSMCs并构建细胞缺氧模型。采用RNA-seq技术对缺氧组(Hypoxia组)和常氧对照组(Control组)的细胞进行转录组测序,筛选出差异表达基因(DEGs)后进行GO富集分析和KEGG通路富集分析,并使用qRT-PCR对测序结果的准确性进行验证。结果模拟高原环境导致小鼠神经元损伤,并且这种损伤伴随脑血管平滑肌收缩功能改变;RNA-seq测序结果显示,Hypoxia组和Control组相比有511个DEGs(变化大于2倍;P<0.05),其中298个上调差异基因,213个下调差异基因。GO富集分析DEGs主要富集在线粒体结构功能以及糖代谢相关途径。在KEGG通路富集分析中,糖酵解/糖异生、氨基酸生物合成、RIG-I样受体信号通路较显著(P<0.05)。结论CVSMCs可通过改变能量代谢、物质代谢、信号传导等多个途径在低气压缺氧脑损伤中发挥作用。本研究为后续进一步研究CVSMCs在低气压缺氧脑损伤中的作用机制提供了新的方向和思路。Objective To investigate the role of cerebrovascular smooth muscle cells(CVSMCs) in hypobaric hypoxic brain injury in mice and its possible mechanism.Methods Male C57BL/6J mice were randomly divided into plain group(Sham group) and plateau group(HA group),with 10 mice in each group.HE staining,immunohistochemistry,immunofluorescence and Western blotting were used to observe brain injury and functional changes of cerebrovascular smooth muscle(CVSM) in each group.Primary CVSMCs of another 20 C57BL/6J mice were isolated and cultured,and a cellular hypoxia model was established.Transcriptome sequencing of hypoxic(Hypoxia group) and normoxic(Control group) cells was performed by RNA-seq technology.Differentially expressed genes(DEGs) were screened for GO enrichment analysis and KEGG pathway enrichment analysis,and qRT-PCR was used to verify the accuracy of sequencing results.Results The simulated high-altitude environment caused neuron damage in mice,and this damage was accompanied by changes in the contractile function of CVSM.RNA-seq results showed that there were 511 DEGs(fold change>2 and P<0.05) in the Hypoxia group compared with the Control group,including 298 up-regulated DEGs and 213 down-regulated DEGs.In GO enrichment analysis,DEGs were mainly enriched in the structure and function of mitochondria and the related pathways of glycometabolism.In KEGG pathway enrichment analysis,glycolysis/gluconeogenesis,biosynthesis of amino acids and RIG-I-like receptor signaling pathways were significantly enriched(P<0.05).Conclusion CVSMCs can play a role in hypobaric hypoxic brain injury by changing energy metabolism,substance metabolism,signal transduction and other ways.This study provides a new direction and idea for further study on the mechanism of CVSMCs in hypobaric hypoxic brain injury.

关 键 词：脑血管平滑肌细胞 低气压缺氧脑损伤 RNA-SEQ GO富集分析 KEGG通路富集分析 

分 类 号：R743[医药卫生—神经病学与精神病学]