环状RNA参与自身免疫性肝炎小鼠肝损伤的相关性分析  

Correlation analysis of circular RNAs involved in liver injury in mice with autoimmune hepatitis

作　　者：侯艺文 刘莹 李竹蓉 陈晨 李振城 刘杨 Hou Yiwen;Liu Ying;Li Zhurong;Chen Chen;Li Zhencheng;Liu Yang(School of Basic Medical Sciences,Shanxi University of Chinese Medicine,Jinzhong 030619,Shanxi Province,China)

机构地区：[1]山西中医药大学基础医学院,山西省晋中市030619

出　　处：《中国组织工程研究》2024年第14期2152-2158,共7页Chinese Journal of Tissue Engineering Research

基　　金：风湿免疫性疾病中西医结合基础研究项目(2023XKJS-03),项目参与人:刘杨;中西医结合防治风湿免疫病山西省科技创新人才重点团队项目(202204051002033),项目参与人:刘杨;山西中医药大学科技创新项目(2022TD2003),项目负责人:刘杨;山西省重点国别科技合作项目(202104041101013),项目负责人:刘杨;山西省高等学校科技创新项目(2020L0423),项目负责人:刘杨;山西中医药大学科技创新能力培育计划(2020PY-JC-07),项目负责人:刘杨。

摘　　要：背景:自身免疫性肝炎的发病机制尚未明确阐明,环状RNA(CircRNAs)是RNA领域的一个研究热点,参与了许多自身免疫性疾病的发病,但CircRNAs在自身免疫性肝炎中的作用尚不清楚。目的:探讨CircRNAs与刀豆蛋白A诱导自身免疫性肝炎小鼠肝损伤的关系。方法:对前期微阵列技术筛选出差异表达的CircRNAs谱进行生物信息学分析,包括基因本体论(GO)及京都基因与基因组百科全书(KEGG)富集分析,探讨上述差异表达基因潜在的生物学功能。采用随机数字表法将12只C57BL/6小鼠分为正常组和模型组,每组6只,模型组通过尾静脉注射刀豆蛋白A建立自身免疫性肝炎模型,造模12 h后处死小鼠,提取小鼠肝脏及外周血,利用qRT-PCR技术验证部分CircRNAs的表达水平,通过比色法检测小鼠血清中肝损伤指标谷丙转氨酶和谷草转氨酶的水平,通过微孔板法检测小鼠肝脏中氧化应激指标丙二醛和NO的水平,并将肝损伤指标和氧化应激水平的相关性进行分析。结果与结论:①GO分析结果显示,表达上调CircRNAs的靶基因参与的生物过程主要为“SNARE复合装配的调节”(P=0.004)等,分子功能主要为“金属离子结合”(P=0.00029)等,主要富集在“CORVET复合体”(P=0.075)等细胞组分;表达下调circRNAs的靶基因参与的生物过程主要为“胰液分泌的负调节”(P=0.00042)等,分子功能主要为“转录激活因子活性”(P=0.025)等,主要富集在“膜外组分”(P=0.006)等细胞组分;②KEGG富集分析结果显示,表达上调CircRNAs的靶基因主要富集于“碱基切除修复”(P=0.026)等信号通路;③与正常组比较,模型组小鼠血清谷丙转氨酶和谷草转氨酶及肝组织中丙二醛和NO的水平升高(P<0.01),mmu-circ-0001520和mmu-circ-0001577的表达升高(P<0.05);④Spearman相关分析显示,mmu-circ-0001520和mmu-circ-0001577的表达与谷丙转氨酶、谷草转氨酶、丙二醛、NO存在正相关关系;⑤结果显示,CircRBACKGROUND:The pathogenesis of autoimmune hepatitis has not been clearly elucidated.Circular RNA(CircRNA)is a research hotspot in the field of RNA and is involved in the pathogenesis of many autoimmune diseases.However,the role of CircRNA in autoimmune hepatitis remains unclear.OBJECTIVE:To investigate the relationship between CircRNA(CircRNA)and concanavalin A induced liver injury in mice with autoimmune hepatitis.METHODS:Bioinformatics analysis was performed on CircRNA profiles selected by previous microarray technology,including gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses,so as to explore the potential biological functions of these differentially expressed genes.Twelve C57BL/6 mice were randomized into normal group and model group(n=6 per group).Autoimmune hepatitis model was established by tail vein injection of concanavalin A in the model group.Mice were killed at 12 hours after modeling to extract mouse liver and peripheral blood.The expression levels of CircRNAs were verified by qRT-PCR.Serum alanine aminotransferase and aspartate aminotransferase levels were detected by colorimetric method.The levels of oxidative stress indexes malondialdehyde and nitric oxide in mouse liver were detected by microplate method.The correlation between oxidative stress level and liver injury index was analyzed.RESULTS AND CONCLUSION:The results of GO analysis showed that the target genes with up-regulated CircRNAs expression were mainly involved in the biological processes of SNARE complex assembly regulation(P=0.004),their molecular functions were mainly metal ion binding(P=0.00029),and the cell components were mainly enriched in CORVET complex(P=0.075).The biological processes involved in the down-regulated circRNAs target genes were mainly“negative regulation of pancreatic secretion”(P=0.00042),the molecular functions were mainly“transcriptional activator activity”(P=0.025),and the cell components were mainly enriched in“extracellular components”(P=0.006).KEGG results

关 键 词：肝损伤 自身免疫性肝炎 刀豆蛋白A 环状RNA(CircRNAs) 相关分析 

分 类 号：R459.9[医药卫生—治疗学] R319[医药卫生—临床医学] R963