四氯化碳诱导的急性肝损伤小鼠肝脏差异表达基因筛选及钙离子转运相关基因动态变化分析  

Screening of Differentially Expressed Genes in the Liver of Mice with Carbon Tetrachloride-induced Acute Liver Injury and Analysis of the Dynamic Changes of Calcium Ion Transport-related Genes

作　　者：曹丽丽 王羚鸿 额尔德木图[1] 布仁其其格[1] 赵林昀 马春丽 包玉龙[1] CAO Lili;WANG Linghong;Eerdemutu;Burenqiqige;ZHAO Linyun;MA Chunli;BAO Yulong(Inner Mongolia Medical University,Hohhot010110,China)

机构地区：[1]内蒙古医科大学,内蒙古呼和浩特010110

出　　处：《畜牧与饲料科学》2024年第6期1-11,共11页Animal Husbandry and Feed Science

基　　金：内蒙古自然科学基金项目(2021MS08029,2024M S08054,2024LHMS08024);内蒙古自治区高等学校青年科技英才支持计划(NJYT23052);内蒙古自治区高等学校创新团队发展计划(NMGIRT2418);内蒙古自治区蒙医药协同创新中心项目(MYYXTPY202401);内蒙古医科大学实验室开放基金项目(2024ZN21);内蒙古自治区“一流学科科研专项项目”(YLXKZX-NYD-009)。

摘　　要：[目的]筛选与四氯化碳(CCl_(4))诱导的肝损伤发生发展相关的基因和关键信号通路,分析肝损伤过程中钙离子(Ca^(2+))转运相关基因表达的动态变化。[方法]将30只8周龄雄性C57BL/6J小鼠随机分为对照组和模型组,每组15只。对照组按0.1 m L/(10 g·BW)的剂量单次腹腔注射生理盐水,模型组按0.1 mL/(10 g·BW)的剂量单次腹腔注射CCl_(4)橄榄油混合物(CCl_(4)∶橄榄油=1∶4,V/V),建立CCl_(4)诱导的急性肝损伤模型。在造模后第2、5、7天,采用ELISA法检测模型组小鼠血清中ALT、AST活力,利用HE染色法和Masson染色法观察肝脏组织病理学变化。从对照组(C组)、造模后第2天组(D2组)、造模后第5天组(D5组)各选取3个生物学重复的肝脏组织样本构建cDNA文库进行转录组测序,对获得的转录组测序数据进行组装及功能注释;使用DESeqR包(1.10.0)筛选显著差异表达基因,利用KOBAS软件进行GO功能注释和KEGG通路富集分析,并对涉及的Ca^(2+)信号通路显著差异表达基因进行分析。[结果]与对照组相比,模型组小鼠肝损伤后第2天血清中ALT、AST活力均极显著(P<0.01)升高。肝损伤后第2天,模型组小鼠肝小叶结构破坏,肝门静脉周围形成大面积坏死与大面积胶原沉积,肝细胞排列紊乱,可见炎性细胞浸润。转录组测序结果显示,D2组与C组相比,差异表达基因总数为6954个,上调和下调基因分别为3577个和3377个;D5组与C组相比,差异表达基因总数为6028个,上调和下调基因分别为3202个和2826个;D5组与D2组相比,差异表达基因总数为6657个,上调和下调基因分别为3200个和3457个;三组之间共有1462个共同变化的差异表达基因。GO功能富集分析显示,D2组与C组之间差异表达基因富集的GO条目主要涉及代谢、信号转导、Ca^(2+)运输等;D5组与C组之间差异表达基因富集的GO条目主要涉及免疫反应、细胞内信号转导;D5组与D2组之间差异表达基因富集的GO条[Objective]The aim of this study was to screen the genes and key signaling pathways associated with the development of carbon tetrachloride(CCl_(4))-induced liver injury,analysing the dynamic changes in the expression of genes related to calcium ion(Ca^(2+))transport during liver injury.[Method]Thirty 8-week-old male C57BL/6J mice were randomly divided into control and model groups,with 15 mice in each group.The control group was given a single intraperitoneal injection of saline at a dose of 0.1 mL/(10 g·BW),and the model group was given a single intraperitoneal injection of a mixture of CCl_(4) as well as olive oil(CCl_(4)∶olive oil=1∶4,V/V)at a dose of 0.1 mL/(10 g·BW)to establish a model of CCl_(4)-induced acute liver injury.The viability of ALT and AST in the serum of mice in the model group was detected by ELISA on days 2,5 and 7 after modelling.Subsequently,the histopathological changes of the liver were observed using HE staining and Masson staining.Three biological replicates of liver tissue samples were selected from each of the control group(Group C),the group on the 2nd day after modelling(Group D2),and the group on the 5nd day after modelling(Group D5)to construct cDNA libraries for transcriptome sequencing,and the obtained data were assembled and functionally annotated.Afterwards,DESeqR package(1.10.0)was used to screen the significantly differentially expressed genes,and KOBAS software was employed to perform GO functional annotation and KEGG pathway enrichment analysis of the significantly differentially expressed genes involved in the Ca^(2+)signalling pathway.[Result]Compared with the control group,ALT and AST activity in serum of mice in the model group on the 2nd day after liver injury were highly significant(P<0.01)increased.On the 2nd day after liver injury,the structure of hepatic lobules in the model group was destroyed.Large necrosis and collagen deposition were formed around the hepatic portal vein,the arrangement of hepatocytes was disordered,and inflammatory cell infiltration cou

关 键 词：肝损伤 转录组测序 Ca2+信号通路 线粒体钙稳态 

分 类 号：R965.1[医药卫生—药理学] R575.3[医药卫生—药学] R114