title: Response of porcine hepatocytes in primary culture to plasma from severe viral hepatitis patients.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Institute of Infectious Diseases, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
authors.forename: Yong-Bo
authors.initials: YB
authors.lastname: Cheng
authors.forename: Ying-Jie
authors.initials: YJ
authors.lastname: Wang
authors.forename: Shi-Chang
authors.initials: SC
authors.lastname: Zhang
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
authors.forename: Zhi
authors.initials: Z
authors.lastname: Chen
authors.forename: Jia-Jia
authors.initials: JJ
authors.lastname: Li
mesh_terms: D000818:Animals; D002374:Catalase / Q000378:metabolism; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D005978:Glutathione / Q000032:analysis; D006525:Hepatitis, Viral, Human / Q000097:blood*; D022781:Hepatocytes / Q000502:physiology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D014176:Protein Biosynthesis; D013552:Swine; D013556:Swine, Miniature
chemical_list: D004247:DNA; D007770:L-Lactate Dehydrogenase; D002374:Catalase; D005978:Glutathione