Screening For Potential Key Mirnas In Human Alcoholic Hepatitis And The Role Of MiR-146a-5p Based On GEO Database

Liver is an important place for alcohol metabolism in human body.When drinking too much for a long time,alcohol and its metabolite acetaldehyde,can cause necrosis and regeneration of hepatocytes repeatedly,which can lead to alcoholic fatty liver,alcoholic hepatitis,liver fibrosis and cirrhosis.When it gets worse,it may even develop into liver cancer.Howerer,there is no targeted drug for alcoholic hepatitis at present,and further research for novel therapeutic targets is also the current hotspot.MiRNAs are involved in many physiological activities.The regulatory mechanism of miRNA on alcoholic hepatitis is helpful for us to develop novel drugs for the treatment of alcoholic hepatitis.After screening expression profile of miRNA and mRNA in alcoholic hepatitis from GEO database,the differentially expressed miRNAs and mRNAs were obtained,then GO enrichment analysis and KEGG pathway analysis for the differentially expressed mRNAs were performed by using David tool on line,Combined with the differentially expressed miRNAs of HBV in GEO database,the potential key miRNAs were selected.The target genes of key miRNAs were predicted by biological software,and the key regulatory network of miRNA-mRNA in alcoholic hepatitis was constructed,and then the GO enrichment analysis and KEGG enrichment analysis were carried out by using David tool on line,.The results showed that the target genes were enriched in 45 biological processes including the positive regulation of GTPase activity and cell activation,and so on.Specially,the potential target genes of miR-146a-5p were involved in some biological processes signaling pathways and,such as protein binding,exosomes and so on.Therefore,we selected miR-146a-5p as one key factor for further research.In this paper,the relative mRNA level of miR-146a-5p in human normal hepatocytes 7702 injured by alcohol was detected by q PCR,and the cell activity of damaged hepatocytes was detected by MTT.The results showed that both cell activity and relative expression level of miR-146a-5p were decreased with the increase of alcohol concentration.In order to explore the protective effect of miR-146 a on hepatocyte,miR-146a-5p mimics and negative control mimics NC were transfected by liposome mediated method.The results showed that the cell activity was increased in cells transfected with miR-146a-5p mimic group when compared with NC + alcohol treatment group,the the expression of the active drug metabolism related protein CYP2E1,SOD and PCNA was increased,on the contrary,the contents of GSH,AST/ALT,SOX5,TNF-α,IL-6 and BCL-2 were decreased.It suggested that the overexpression of miR-146a-5p could reduce the damage caused by alcohol on hepatocyte.In conclusion,the key miRNAs were selected by the combined analysis of miRNAs and mRNAs in human alcoholic hepatitis from GEO,it was verified that the overexpression of miR-146a-5p could reduce the alcohol damage to hepatocytes by liposome transfection.These results lay a foundation for the further development of targeted drugs for the treatment of alcoholic hepatitis.