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
- VernacularTitle: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
- Author:
Di GUO
1
;
Ying LIU
1
;
Yang LIU
1
Author Information
- Publication Type:Journal Article
- Keywords: Hepatitis,Autoimmune; RNA,Circular; MicroRNAs; RNA,Messenger; Mice,Inbred C57BL
- From: Journal of Clinical Hepatology 2025;42(5):878-887
- CountryChina
- Language:Chinese
- 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.
