Integrated Analysis Of Glycogenes Expression And Glycan Profiles In Mice With Liver Fibrosis

Aims:Protein glycosylation is one of the major posttranslational modification, which an important effect on protein folding, stability and activity. It has previously been shown that altered of glycosylation involved in various biological processes such as cell adhesion, pathogen recognition and metastasis. Hepatic fibrosis is considered a model of the wound-healing response to liver injury. Without appropriate treatment, it would develop cirrhosis, and have a risk to develop hepatocellular carcinoma. The aim of this study was to investigate potential associations between glycogenes expression and glycan profiles to evaluate hepatic fibrosis in a mouse model which induced by CC14.Method: Liver fibrosis was induced by CC14intraperitoncal injection in mouse models. Liver tissues were separated and used to isolate RNA, and then, targets were prepared for hybridized of mouse whole-genome oligonucleotide microarray.The differential expression glycogenes were filtrated after analysis, and the results were verified by RT-PCR. For another, proteins were extractd from liver tissue with fibrosis and normal controls respectively. After labeled with Cy3fluorescence, the labeled proteins were incubated with lectin microarray. The data was normalized by global method, and abnormal glycopatterns which associated with fibrosis were screened. Lectin histochemistry was used to confirm any altered glycopatterns. The expression level of glycosyltransferases which involved in the synthesis of abnormal glycopatterns was analysis by Western Blot. Integrate these results, our finding revealed a synthesis pathway of abnormal glycopatterns associated with fibrosis.Result:The result of mouse whole-genome microarray showed that (1) There are1079±44transcripts up-regulated (fold change>1.5, p value<0.05) and579±10transcripts down-regulated (fold change<0.67, p value<0.05) in fibrotic group when compared with controls.(2)227glycogenes which were contained in the microarray, in which,8glycogenes were up-regulated and2were down-regulated in fibrotic group.(3)6glycogenes which transcription levels were altered were verified by RT-PCR, and result was consistant with microarray. The result of Western Blot revealed that the transcription level of and expression level of glycosyltransferase/glycosidase had same altered tendency. The results of lectin microarray indicated that (1) the glycopatterns which recognized by10lectins has higher abundance in fibrosis group, besides, the glycopattern which recognized by lectin STL has higher abundance in normal control group.(2) The result of lectin histochemistry was consistant with lectin arrays’.Conclusion:1. In process of liver fibrosis, the transcription level of glycosyltransferase/glycosidase which involved in modification of glycosylation on protein were altered, and it has an effect on modification of glycosylation on glycoprotein in hepatocyte.2. Integrated analysis of results of gene expression profile microarray, Western Blot and lectin array, our finding revealed that the synthetic pathway of " Tn antigenâ†'T antigen (core-1)â†'sialyl-T antigen" was activated for O-glycan during the process of liver fibrosis.