Functional proteomic analysis of nonalcoholic fatty liver disease in rat models

• DOI: 10.1002/hep.23486
• Publication Date: 2009-12-04T15:19:02Z
• Authors: Xuequn Zhang; Juntao Yang; Yuanbiao Guo; Hua Ye; Chaohui Yu; Chengfu Xu; Lei Xu; Songfeng Wu; Wei Sun; Hangdong Wei; Xue Gao; Yunping Zhu; Xiaohong Qian; Ying Jiang; Youming Li; Fuchu He
• Keywords: Steatosis

Nonalcoholic fatty liver disease (NAFLD) has emerged as a common public health problem that can progress to end-stage disease. A high-fat diet (HFD) may promote the development of NAFLD through mechanism is poorly understood. We adopted proteomic approach examine effect HFD on proteome during progression NAFLD. Male Sprague-Dawley rats fed an for 4, 12, and 24 weeks replicated human NAFLD: steatosis, nonspecific inflammation, steatohepatitis. Using two-dimensional difference gel electrophoresis (DIGE) combined with matrix-assisted laser desorption ionization time flight/time flight analysis, 95 proteins exhibiting significant changes (ratio ≥ 1.5 or ≤−1.5, P < 0.05) were identified. Biological functions these reflected phase-specific characteristics The potential role enoyl–coenzyme hydratase (ECHS1), enzyme catalyzes second step mitochondrial acid beta-oxidation, received further investigation. First, reduced protein level ECHS1 was validated both in rat models patients biopsy-proven hepatic simple steatosis via immunoblotting immunohistochemical analysis. Then small interfering RNA (siRNA)–mediated knockdown murine hepatocyte cell line alpha mouse 12 (AML12) demonstrated increased cellular lipid accumulation induced by free (FFA) overload. Furthermore, using hydradynamic transfection method, vivo silencing siRNA duplexes targeting investigated mice. Administering specifically expression mice liver, which significantly exacerbated HFD. Conclusion: Our results revealed down-regulation contributed HFD-induced help clarify pathogenesis point targets therapeutic interventions. (HEPATOLOGY 2010.)