甘草酸介导miRNAs对大鼠酒精性肝损伤的保护作用

Glycyrrhizic acid-mediated protective effect of miRNAs on alcoholic liver injury in rats

目的探讨miRNAs在甘草酸治疗大鼠酒精性肝损伤过程中的作用及其机制。方法将45只雄性SD大鼠随机分成甘草酸组(灌胃56%白酒和甘草酸)、模型组(灌胃56%白酒)和对照组(灌胃蒸馏水),给药8周后采集血液和肝组织。检测血清天冬氨酸氨基转移酶(AST)及丙氨酸氨基转移酶(ALT)水平;提取肝组织RNA,大鼠miRNA芯片检测,对miRNA表达量进行分析,对差异miRNA进行靶基因预测,并采用Gene Ontology(GO)和KEGG Pathway富集分析了解差异miRNA的功能,使用Cytoscape构建差异miRNA-mRNA-Pathway调控网络进一步筛选调控关键miRNA与通路,qRT-PCR对挑选的miRNA进行表达量验证分析。结果与模型组相比,甘草酸组能改善肝组织病变,降低肝血清AST和ALT水平(P<0.05)。通过比较分析甘草酸组和模型组芯片数据,共筛选出差异表达miRNA 13个(P<0.05,fold change≥1.5),其中上调10个,下调3个。差异miRNA靶基因GO分类注释显示,差异miRNA主要与细胞黏附、抗氧化活性、代谢过程、生物过程调控、细胞杀伤、免疫系统等功能相关。差异miRNA靶基因KEGG Pathway通路分析显示,MAPK信号通路、mTOR信号通路、Ras信号通路、Hippo信号通路、PI3K-Akt信号通路、wnt信号通路、细胞凋亡等信号通路可能在甘草酸改善酒精性肝损伤病变过程中发挥着重要的调控作用。结论该研究建立了甘草酸治疗大鼠酒精性肝损伤的miRNA表达谱,提示miR-615、miR-107-3p和miR-292-5p可能在甘草酸治疗酒精性肝损伤过程中起着重要的作用。

Objective To investigate the mechanism of miRNAs in glycyrrhizic acid treatment of alcohol-induced liver injury in rats.Methods 45 male SD rats were randomly divided into glycyrrhizin group,model group and control group.The rats in glycyrrhizin group were given 56%liquor and glycyrrhizin,the rats in model group were given 56%liquor,and the rats in control group were given distilled water for 8 weeks.The blood was collected and the serum was separated by centrifugation to detect the levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT).RNA were extracted from liver tissues,miRNAs were detected by rat miRNA microarray,and their expression levels were analyzed.The miRNA target genes of differential miRNA were predicted.Gene Ontology(GO)and KEGG Pathway enrichment analysis were used to understand the function of differential miRNA,and the differential miRNA-mRNA-Pathway regulatory network was constructed using Cytoscape to further screen the regulatory key miRNA and key pathways.qRT-PCR was used to verify the expression of selected miRNA.Results Compared with the model group,the glycyrrhizin group could significantly improve the liver tissue lesions,and reduce the liver serum AST and ALT levels(P<0.05).Compared the microarray data of glycyrrhizin group and model group,a total of 13 differentially expressed miRNA were screened(P<0.05,fold change≥1.5),of which 10 were up-regulated and 3 were down-regulated.The GO classification annotation of differential miRNA target genes showed that differential miRNA were related to cell adhesion,antioxidant activity,metabolic process,biological process regulation,cell killing,immune system and other functions.The KEGG Pathway analysis of differential miRNA target genes showed that MAPK signaling pathway,mTOR signaling pathway,Ras signaling pathway,Hippo signaling pathway,PI3K-Akt signaling pathway,wnt signaling pathway,apoptosis and other signaling pathways might play an important regulatory role in the improvement of alcoholic liver injury by glycyrrhic acid.Conclusion This study established the miRNA expression profile of glycyrrhizin in the treatment of alcoholic liver injury in rats,suggested that miR-615,miR-107-3p and miR-292-5p might play an important role in the treatment of alcoholic liver injury by glycyrrhizin.