Viral Hepatitis
Copyright ©2007 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jul 7, 2007; 13(25): 3472-3477
Published online Jul 7, 2007. doi: 10.3748/wjg.v13.i25.3472
Topological and evolutional relationships between HCV core protein and hepatic lipid vesicles: Studies in vitro and in conditionally transgenic mice
Ming-Ling Chang, Jeng-Chang Chen, Chau-Ting Yeh, I-Shyan Sheen, Dar-In Tai, Ming-Yu Chang, Cheng-Tang Chiu, Deng-Yn Lin, D Montgomery Bissell
Ming-Ling Chang, Chau-Ting Yeh, I-Shyan Sheen, Dar-In Tai, Cheng-Tang Chiu, Deng-Yn Lin, Liver Research Center and Department of Hepatogastroenterology, Chang Gung Memorial Hospital, Chang Gung University, College of Medicine, Taoyuan, Taiwan, China
Ming-Ling Chang, D Montgomery Bissell, Liver Center and Department of Medicine, University of California, San Francisco, United States
Jeng-Chang Chen, Department of Surgery, Chang Gung Children's Hospital, Taoyuan, Taiwan, China
Ming-Yu Chang, Division of Pediatric Neurology, Department of Pediatrics, Chang Gung Children's Hospital at Linkou, Chang Gung University, College of Medicine, Taoyuan, Taiwan, China
Author contributions: All authors contributed equally to the work.
Supported by grants from National Science Council, Taiwan, China. No. NSC 93-2314-B-182A-148, 94-2314-B-182A-185 and 95-3112-B-182A-002 and Chang Gung Memorial Hospital, Taoyuan, Taiwan, China. No. CMRPG 33014, CMRPG 340341 and SMRPG350081
Correspondence to: Ming-Ling Chang, Liver Research Unit, Department of Hepatogastroenterology, Chang Gung Memorial Hospital, No 5, Fu Hsing Street, Kuei Shan, Taoyuan, Taiwan, China. kitty8210@yahoo.com
Telephone: +886-3-3281200-8102 Fax: +886-3-3272236
Received: April 5, 2007
Revised: April 7, 2007
Accepted: April 26, 2007
Published online: July 7, 2007
Abstract

AIM: To investigate a specific association between hepatic steatosis and hepatitis C virus (HCV) core.

METHODS: HeLa cells and primary mouse hepatocytes were transfected with HCV core plasmid, and conditional transgenics in which hepatic over-expression of HCV core is regulated by the tetracycline-off system, were developed. The expression of the HCV core was assessed over one to six months after withdrawal of doxycycline (dox) by immunohistochemistry (IHC) and Western blotting and by sequential liver biopsy. Hepatic steatosis was evaluated using oil red stain. 8-hydroxydeoxyguanosine (8-OHdG) stains and caspase levels were conducted to clarify hepatic oxidative stress and apoptosis rate. Serum aminotransferase was checked.

RESULTS: The transfected hepatocytes had globular cores under the lipid vesicles. In transgenic mice on control diet, core expression was robust, localized to the cytoplasmic vesicle membrane and strongly associated with microvesicular steatosis, which was gradually replaced by macrovesicular steatosis. However, both steatosis and core positive hepatocytes diminished with time. Increases in aminotransferase, caspase and 8-OHdG were associated with peak core expression.

CONCLUSION: The core protein was readily detected and morphologically associated with steatosis in individual hepatocytes both in vitro and in vivo. In vivo, oxidative stress caused by the core potentially reduced the number of core positive hepatocytes and in parallel the level of steatosis. To our knowledge, this is the first animal model that directly shows topological relationship between HCV core and hepatic lipid vesicles.

Keywords: Hepatic steatosis; Conditional transgenic mice; HCV core; Oxidative stress