基于微阵列数据的circRNA-miRNA-mRNA竞争性内源RNA网络构建及其与自身免疫性肝炎小鼠模型肝损伤的相关性分析
Construction of a circRNA-miRNA-mRNA competitive endogenous RNA network based on microarray data and its correlation with liver injury in a mouse model of autoimmune hepatitis
摘要
Abstract
Objective To construct a circRNA-miRNA-mRNA competitive endogenous RNA(ceRNA)network,to investigate its potential regulatory mechanism in a mouse model of autoimmune hepatitis(AIH)induced by concanavalin A(ConA),and to verify the association between the expression of key genes and liver injury.Methods High-throughput data were used to identify differentially expressed circRNAs,miRNAs,and mRNAs,and the Pearson correlation analysis and the Miranda program were used to predict the pairing relationships between miRNAs and mRNAs/circRNAs and construct a ceRNA network.The gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed for the differentially expressed genes in the network.A total of 12 specific pathogen-free male C57BL/6 mice were divided into control group and model group using a random number table,with 6 mice in each group.The mice in the model group were given injection of ConA via the caudal vein to establish a mouse model of AIH,and those in the control group were given injection of normal saline.The methods of qRT-PCR and Western blot were used to validate the expression levels of circ_0001577,miR-7055-3p,and Akt3.The serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were measured,as well as the content of malondialdehyde(MDA)and nitric oxide(NO)in liver tissue,and their correlation with gene expression was analyzed.The independent-samples t test was used for comparison of continuous data between two groups,and the Spearman correlation analysis was used to investigate the correlation between gene expression levels and liver injury indicators.Results A ceRNA network containing 5 795 circRNA-miRNA-mRNA pairings was constructed,and circ_0001577 was identified as the central gene.Compared with the control group,the model group had significant increases in the expression levels of circ_0001577 and Akt3 and a significant reduction in the expression of miR-7055-3p(all P<0.05),and circ_0001577 was positively correlated with Akt3(r=0.861,P<0.001),while miR-7055-3p was negatively correlated with circ_0001577 and Akt3(r=-0.644 and-0.855,both P<0.05).Compared with the control group,the model group had a significantly higher protein expression level of Akt3 in the liver(1.04±0.10 vs 0.72±0.06,t=-6.49,P=0.001),which was positively correlated with circ_0001577(r=0.579,P=0.048)and was negatively correlated with miR-7055-3p(r=-0.891,P<0.001).Compared with the control group,the model group had significant increases in the serum levels of ALT and AST and the content of MDA and NO in liver tissue(all P<0.05),and these liver injury indicators were positively correlated with circ_0001577 and Akt3(r=0.849,0.865,0.811,0.801;0.889,0.954,0.938,and 0.961,all P<0.05)and were negatively correlated with miR-7055-3p(r=-0.687,-0.818,-0.833,and-0.870,all P<0.05);in addition,they were positively correlated with the protein expression level of Akt3(r=0.648,0.796,0.848,and 0.860,all P<0.05).Conclusion This study shows that circ_0001577 promotes the expression of Akt3 by competitively adsorbing miR-7055-3p and relieving the inhibition of Akt3,thereby participating in the development and progression of AIH.关键词
肝炎,自身免疫性/RNA,环状/微RNAs/RNA,信使/小鼠,近交C57BLKey words
Hepatitis,Autoimmune/RNA,Circular/MicroRNAs/RNA,Messenger/Mice,Inbred C57BL引用本文复制引用
郭地,刘莹,刘杨..基于微阵列数据的circRNA-miRNA-mRNA竞争性内源RNA网络构建及其与自身免疫性肝炎小鼠模型肝损伤的相关性分析[J].临床肝胆病杂志,2025,42(5):878-887,10.基金项目
国家中医药管理局高水平中医药重点学科建设项目(2024) （2024）
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(202204051002033) （202204051002033）
山西省科学技术厅山西省重点国别科技合作项目(202104041101013) （202104041101013）
山西省应用基础研究计划青年科学基金项目(202203021222272) （202203021222272）
山西省中医药管理局科研项目(2024ZYYAD008) （2024ZYYAD008）
山西中医药大学科技创新团队项目(2022TD2003) （2022TD2003）
中西医结合治疗风湿免疫疾病重点实验室(zyyyjs2024021) The Construction Project of High-level Traditional Chinese Medicine Key Discipline of National Administration of Traditional Chinese Medicine(2024) （zyyyjs2024021）
Key Team of Scientific and Technological Innovation Talents of Shanxi Province with Integrated Traditional Chinese and Western Medicine for Preventing and Treating Rheumatological Diseases(202204051002033) （202204051002033）
Special Project with Important Nations of Scientific and Technological Cooperation and Exchange of Shanxi Province(202104041101013) （202104041101013）
Youth Science and Technology Research Fund of Applied Basic Research Program of Shanxi Province(202203021222272) （202203021222272）
Scientific Research Project of Administration of Traditional Chinese Medicine in Shanxi Province(2024ZYYAD008) （2024ZYYAD008）
Science and Technology Innovation Project of Shanxi University of Chinese Medicine(2022TD2003) （2022TD2003）
Key Laboratory of Rheumatological and Immunological Diseases Treated by Integrated Chinese and Western Medicine(zyyyjs2024021) （zyyyjs2024021）
