| Objective:In order to explore the potential molecular mechanism of pathogenesis of spontaneous type 2 diabetes mellitus(T2DM)in Chinese hamsters,we used tandem mass tags(TMT)combined with liquid chromatography-tandem mass spectrometry(LC-MS/MS)to identify and analyze the adipose tissue of T2 DM Chinese hamsters.Key differentially expressed proteins(DEPs)and KEGG signaling pathways were expected to be associated with the pathogenesis of T2 DM Chinese hamsters.Methods:Spontaneous T2 DM Chinese hamsters with fasting blood glucose ≥ 6 mmol/L were selected as the diabetic group,and hamsters with fasting blood glucose ≤ 4.5 mmol/L were selected as the normal control group,and take adipose tissue for TMT quantitative proteomics analysis.GO and KEGG were used to perform functional enrichment analysis on the identified DEPs.Afterwards,use western blotting to verify the selected DEPs,observe the pathological changes of Chinese hamster adipose tissue by HE staining,quantitatively detect the number of peroxisomes in the adipose tissue by immunofluorescence technology.Finally,use the kit to determine the activity of glucose-6-phosphate(G6PDH)and 6-phosphogluconate dehydrogenases(6PGDH)in adipose tissue,and the content of triglyceride(TG),non-esterified fatty acid(NEFA),superoxide dismutase(SOD),glutathione peroxidase(GDH-PX)and malondialdehyde(MDA)in serum.Results:A total of 212 DEPs(125 up-regulated and 87 down-regulated)were identified in adipose tissue by quantitative proteomic analysis of TMT.The GO and KEGG functional enrichment analysis showed that some DEPs were gathered in the peroxisome pathway,including PEX11 A and PEX11 B,which affect the peroxisome division,and ACOX1,HSD17B4,EHHADH and ACAA1,which affect the peroxisome fatty acid β-oxidation,were down-regulated in type 2 diabetic hamlets.Western blotting results showed that the expression level of the candidate DEPs in adipose tissue decreased significantly.Histopathological results showed that the adipocytes of T2 DM hammers were significantly larger than that of the normal group,and fluorescence staining results showed that the number of peroxisomes in adipocytes was decreased.The activity of G6 PDH and 6PGDH in adipose tissue is weakened.In serum,the contents of TG and NEFA increased,SOD and GSH-PX decreased,and MDA increased.Conclusion:In this study,we used quantitative proteomics to identify the proteins in the adipose tissue of T2 DM Chinese hamsters,and through GO and KEGG databases to perform a functional enrichment analysis of all DEPs.It was found that some DEPs are aggregated in the peroxisome pathway,and the down-regulated DEPs affect the biosynthesis of the peroxisome.This change further affected the lipid oxidation function and ROS scavenging capacity of T2 DM Chinese hamsters,which may have contributed to the development of T2 DM.Our study revealed the poential role of peroxisome in the pathogenesis of T2 DM,providing valuable information for further understanding of the molecular mechanism of the pathogenesis in T2 DM Chinese hamsters. |
PDF Full Text Request