急性脑出血模型大鼠脑组织中差异表达基因测序的验证及功能分析  

Sequencing,verification and functional analysis of differentially expressed genes in brain tissue of a rat model with acute intracerebral hemorrhage

作　　者：高玉广[1] 钟洁[1] 黄德庆[1] 马玉娟 廖煜雄 刘琦琦 Gao Yuguang;Zhong Jie;Huang Deqing;Ma Yujuan;Liao Yuxiong;Liu Qiqi(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西中医药大学第一附属医院,广西壮族自治区南宁市530023

出　　处：《中国组织工程研究》2024年第20期3182-3189,共8页Chinese Journal of Tissue Engineering Research

基　　金：广西教育厅高校中青年教师科研基础能力提升项目(2020KY07015),项目负责人:高玉广;广西壮族自治区卫生健康委员会自筹经费科研课题(20210943),项目负责人:高玉广;广西中医药重点学科建设项目(GZXK-Z-20-47);广西医疗卫生重点培育学科建设项目(桂卫科教发[2021]8号)。

摘　　要：背景:急性脑出血中存在差异表达的基因,这些基因与脑出血发生发展有关。目的:筛选急性脑出血大鼠模型脑组织中差异表达的基因和关键基因,并采用qPCR进行验证,分析关键基因与脑出血后神经功能、脑组织含水量的关系。方法:78只SD大鼠随机分为2组:脑出血组大鼠于右侧尾状核部位注射胶原酶构建脑出血模型,假手术组大鼠于相同部位注射等量生理盐水。运用TRIzol法将2组大鼠的脑组织提取出RNA,转录组测序,筛选急性脑出血脑组织的差异表达的基因,经过qPCR验证,分析基因与脑出血后神经功能、脑组织含水量的关系,并结合生物信息学对关键基因进行GO、KEGG功能富集分析。结果与结论:①筛选出10个关键基因,分别是CXCL8、SERPINE1、TFPI2、CXCR4、GDA、KCNQ5、ERICH3、SCN3B、CACNA1E、CCL20;②脑出血组的关键基因GDA、KCNQ5、ERICH3、SCN3B、CACNA1E含量低于假手术组(P<0.05),CXCL8、SERPINE1、TFPI2、CXCR4、CCL20含量高于假手术组(P<0.05);③关键基因GDA、KCNQ5、ERICH3、SCN3B、CACNA1E含量与脑组织含水量、神经功能缺损评分正相关(P<0.05),CXCL8、SERPINE1、TFPI2、CXCR4、CCL20含量与脑组织含水量、神经功能缺损评分负相关(P<0.05);④GO分析显示基因在生物过程差异表达基因主要富集于白细胞趋化性、趋化因子介导的信号通路;细胞组成差异表达基因主要富集于阳离子通道复合物、离子通道复合物;分子功能差异表达基因主要富集于门控通道活动、离子通道活动;⑤KEGG分析示基因集中于TNF信号通路、谷氨酸能突触、GABA能突触;⑥结论:在脑出血中出现差异表达的基因为CXCL8、SERPINE1、TFPI2、CXCR4、GDA、KCNQ5、ERICH3、SCN3B、CACNA1E、CCL20,这些基因与脑出血后脑组织含水量、神经功能相关,这些基因主要富集在细胞组分、结合功能、细胞突起等相关的生物学功能上。BACKGROUND:There are differentially expressed genes in acute intracerebral hemorrhage,which are related to the occurrence and development of intracerebral hemorrhage.OBJECTIVE:To screen differentially expressed genes and key genes in brain tissue of a rat model with acute intracerebral hemorrhage,to validate them through qPCR,and to analyze the relationships between key genes and the neurological function and brain tissue water content after intracerebral hemorrhage.METHODS:Seventy-eight Sprague-Dawley rats were randomly divided into two groups:in intracerebral hemorrhage group,a rat model of acute intracerebral hemorrhage was made using collagenase injection at the right caudate nucleus;and in sham-operated group,rats were injected with equal amount of saline at the same site.RNA was extracted from rat brain tissues of both groups using the TRIzol method and transcriptome sequencing technology was used to identify differentially expressed genes in brain tissues of acute intracerebral hemorrhage,which were then verified by qPCR and analyzed for the relationships between the genes and neurological function and brain tissue water content after intracerebral hemorrhage.And the key genes were analyzed by GO and KEGG functional enrichment analysis in combination with bioinformatics.RESULTS AND CONCLUSION:Ten key genes were identified,including CXCL8,SERPINE1,TFPI2,CXCR4,GDA,KCNQ5,ERICH3,SCN3B,CACNA1E,and CCL20.The contents of GDA,KCNQ5,ERICH3,SCN3B,and CACNA1E in the intracerebral hemorrhage group were lower than those in the sham-operated group(P<0.05).The contents of CXCL8,SERPINE1,TFPI2,CXCR4 and CCL20 in the intracerebral hemorrhage group were higher than those in the sham-operated group(P<0.05).The contents of GDA,KCNQ5,ERICH3,SCN3B,and CACNA1E were positively correlated with brain tissue water content and neurologic deficit score(P<0.05),while the contents of CXCL8,SERPINE1,TFPI2,CXCR4 and CCL20 were negatively correlated with brain tissue water content and neurologic deficit score(P<0.05).GO analysis indicated

关 键 词：脑出血 基因 差异性表达 神经功能 脑组织含水量 生物信息学分析 

分 类 号：R496[医药卫生—康复医学] R318[医药卫生—临床医学] R446