Viral Hepatitis
Copyright ©2005 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Dec 28, 2005; 11(48): 7585-7590
Published online Dec 28, 2005. doi: 10.3748/wjg.v11.i48.7585
Response of porcine hepatocytes in primary culture to plasma from severe viral hepatitis patients
Yong-Bo Cheng, Ying-Jie Wang, Shi-Chang Zhang, Jun Liu, Zhi Chen, Jia-Jia Li
Yong-Bo Cheng, Ying-Jie Wang, Shi-Chang Zhang, Jun Liu, Zhi Chen, Jia-Jia Li, Institute of Infectious Diseases, Southwest Hospital, Third Military Medical University, Chongqing 400038, China
Author contributions: All authors contributed equally to the work.
Supported by National Natural Science Foundation of China, No. 30470458
Correspondence to: Ying-Jie Wang, MD, Institute of Infectious Diseases, Southwest Hospital, Third Military Medical University, Chongqing 400038, China. wangyj103@263.net
Telephone: +86-23-68754479-8062
Received: April 14, 2005
Revised: June 23, 2005
Accepted: June 24, 2005
Published online: December 28, 2005
Abstract

AIM: To observe the effects of plasma from patients with severe viral hepatitis (SVHP) on the growth and metabolism of porcine hepatocytes and the clinical efficiency of bioartificial liver device.

METHODS: Hepatocytes were isolated from male porcines by collagenase perfusion. The synthesis of DNA and total protein, leakages of AST and LDH, changes in glutathione (GSH), catalase and morphology of porcine hepatocytes exposed to SVHP were investigated to indicate the effect of plasma from patients with severe hepatitis on the growth, injury, detoxification, and morphology of porcine hepatocytes.

RESULTS: The synthesis of DNA and protein was inhibited in the medium containing 100% SVHP compared to the controls. The leakages of LDH and AST increased in porcine hepatocytes following exposure to 100% SVHP for 5 h. The difference between 100% SVHP and 10% newborn calf serum (NCS) was significant in t-test (LDH: t = 24.552, P = 0.001; AST: t = 4.169, P = 0.014). After exposure to SVHP for 24 h, alterations in GSH status were significant (F = 2.746, P<0.05) between porcine hepatocytes in 100% SVHP and 10% NCS, but no alteration occurred in the culture medium after 48 h (F = 4.378, P<0.05). A similar profile was observed in catalase activity. Many round vacuoles were observed in porcine hepatocytes cultured in SVHP. The membranes of these cells became indistinct and almost all the cells died on d 5.

CONCLUSION: Plasma from patients with severe hepatitis inhibits the growth, injures membrane, disturbs GSH homeostasis and induces morphological changes of porcine hepatocytes. It is suggested that SVHP should be pretreated to reduce the toxin load and improve the performance of porcine hepatocytes in extracorporeal liver-support devices.

Keywords: Bioartificial liver; Porcine hepatocytes; Cell culture; Plasma toxicity