Response of porcine hepatocytes in primary culture to plasma from severe viral hepatitis patients.
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.
Cheng
Yong-Bo
YB
Institute of Infectious Diseases, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
Wang
Ying-Jie
YJ
Zhang
Shi-Chang
SC
Liu
Jun
J
Chen
Zhi
Z
Li
Jia-Jia
JJ
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
D004247:DNA; D007770:L-Lactate Dehydrogenase; D002374:Catalase; D005978:Glutathione