Study On Screening And Function Of LncRNAs In Alcohol-Induced Liver Injury

[Objective]Alcoholic liver disease（ALD）is still one of the world’s leading health problems.Studies have shown that long noncoding RNA（lnc RNA）plays an important role in the pathogenesis of ALD.However,the role of lnc RNAs in the pathogenesis of ALD is not understood completely.In this study,we established mouse model of alcoholic liver disease and using gene chip technology to screen differential expression of lnc RNAs.Analyzing the correlation between the disease and lnc RNAs,as well as,the co-expression of lnc RNA and m RNA,an potential lnc RNA had been found out.Using Cis-target gene regulating method,the potential lnc RNA targeting gene had been tried to predict and figured out its function and possible mechanism.[Methods]The 1^st Experiment1.Using chronic alcoholic gavage,established an alcohol-induced liver injury animal model,measured the model from several aspects below:general conditions of the mice,the weight,and the fatal ratio.2.Furthermore,confirmed the model being well done from serum biochemistry markers,histology changes in mice liver.The 2^nd Experiment1.Using microarray technique,figured out the different expression between lnc RNAs and m RNAs in mice hepatocytes.Besides that,perform the microarray analyzing,GO enrichment analyzing and KEGG pathway enrichment analyzing.2.Through the differential expression analyzing and co-expression analyzing of lnc RNA-m RNA,screened 10 lnc RNAs,and select one which is up-regulated to study furthermore.3.Predicted the targeting gene of the lnc RNA through using Cis-regulated gene,and found out the function of targeting gene in alcohol-induced liver injury model.The 3^rd Experiment1.Measured cytokines changing in mice serum by using CBA flow cytometry.2.Using flow cytometry,detected mouse hepatocyte apoptosis ratio in high-dose alcohol-fed mice group and interfering groups.[Results]1.Observed the levels of ALT,AST,GLOB,and TP in serum of high-dose ethanol fed mice groups were elevated（P<0.05）,and the level of ALT was reduced（P<0.05）,through examing serum biochemical markers,staining tissue with HE,and using CBA flow cytometric technology.Hepatocyte degeneration and inflammatory cell infiltration occured in ethanol fed mice liver.The level of IL-6 in serum cytokines was increasing （P<0.05）.All changes above demonstrated that ALD animal models were made successfully.2.Using gene chip technology,we screen out some lnc RNAs in the alcohol-induced injury liver which their expression is up-regulated or down-regulated.3.Screened a up-regulated gene lnc RNA-NONMMUT007193 to focus through the differential expression analyzing and co-expression analyzing of lnc RNA-m RNA.4.Predicted the targeting gene for lnc RNA-NONMMUT007193 is m RNA-Tmtc2.by Cis-regulated targeting genes.5.Detected hepatocyte apoptosis using FCM method（Flow Cytometry）and CBA method （Cytometric Bread Array）.The level of IL-6 in serum of high-dose mice increasesignificantly.The decrease of apoptosis in hepatocytes in high-dose group with si RNA interfering may be related to the lnc RNA-NONMMUT007193.It is suggested that lnc RNA-NONMMUT007193 may play the role of promoting apoptosis in alcohol-induced liver injury.[Conclusions]1.Established an alcohol-induced injury liver model in mouse successfully,and proved it.2.Screened out a number of lnc RNAs in the liver of mice which their expressions were up-regulated or down-regulated.3.A up-regulated gene lnc RNA-NONMMUT007193 is screened out through the differential expression analyzing and co-expression analyzing of lnc RNA-m RNA.4.Used Cis-regulated targeting gene technique to predict the targeting gene for lnc RNA-NONMMUT007193 is m RNA-Tmtc2.5.The increasing of IL-6 in serum of high-dose alcohol-fed mice is probably related to the activation of inflammatory factors after high-dose alcohol-fed.6.lnc RNA-NONMMUT007193 may play the role of promoting apoptosis in alcohol-induced liver injury.