摘要
AIM: To explore mitochondrial dysfunction in nonalcoholic steatohepatitis (NASH) by analyzing the proteome of liver mitochondria from a NASH model.
AIM:To explore mitochondrial dysfunction in nonalcoholic steatohepatitis(NASH)by analyzing the proteome of liver mitochondria from a NASH model.METHODS:The NASH rat model was established by feeding rats a fat-rich diet for 24 wk and was confirmed using hematoxylin and eosin staining of liver tissue and by changes in the levels of serum alanine transaminase,aspartate aminotransferase,triglyceride,total cholesterol and other markers.Liver mitochondria from each group were isolated using differential centrifugation.The mitochondrial samples were lyzed,purified and further analyzed using two-dimensional electrophoresis combined with matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry.Bioinformatic analyses of assigned gene ontology and biological pathway was used to study functional enrichments in the abundant proteomic data.RESULTS:Eight up-regulated and sixteen down-regulated proteins were identified that showed greater than1.5-fold differences between the controls and the NASH group.These dysregulated proteins were predicted to be involved in different metabolic processes including fatty acidβ-oxidation processes,lipid metabolic processes,cell-cycle arrest,cell polarity maintenance,and adenosine triphosphate/sex hormone metabolic processes.Novel proteins that may be involved in NASH pathogenesis including the trifunctional enzyme Hadha,thyroxine,prohibitin,aldehyde dehydrogenase ALDH1L2,UDP-glucuronosyltransferase 2B31,and carbamoyl-phosphate synthase were identified using bioinformatics tools.The decreased expression of Hadha in NASH liver was verified by Western blotting,which was used as a complementary technique to confirm the proteomic results.CONCLUSION:This novel report on the liver mitochondrial proteome of a NASH model may provide a reservoir of information on the pathogenesis and treatment of NASH.
基金
Supported by National Natural Science Foundation of China No.81000169,No.81100277,No.81370008,and No.81200284
the Excellent Young Investigator Foundation of the Health Bureau of Zhejiang Province No.2010QNA011
the Excellent Young Investigator Natural Science Foundation of Zhejiang Province No.R2110159
the Project of Zhejiang Traditional Chinese Medicine Administration Bureau No.2010ZA065
the Fundamental Research Funds for the Central Universities No.2013QNA702
