Effects of plasma from patients with acute on chronic liver failure on function of cytochrome P450 in immortalized human hepatocytes.
BACKGROUND
The bioartificial liver is anticipated to be a promising alternative choice for patients with liver failure. Toxic substances which accumulate in the patients' plasma exert deleterious effects on hepatocytes in the bioreactor, and potentially reduce the efficacy of bioartificial liver devices. This study was designed to investigate the effects of plasma from patients with acute on chronic liver failure (AoCLF) on immortalized human hepatocytes in terms of cytochrome P450 gene expression, drug metabolism activity and detoxification capability.


METHODS
Immortalized human hepatocytes (HepLi-2 cells) were cultured in medium containing fetal calf serum or human plasma from three patients with AoCLF. The cytochrome P450 (CYP3A5, CYP2E1, CYP3A4) expression, drug metabolism activity and detoxification capability of HepLi-2 cells were assessed by RT-PCR, lidocaine clearance and ammonia elimination assay.


RESULTS
After incubation in medium containing AoCLF plasma for 24 hours, the cytochrome P450 mRNA expression of HepLi-2 cells was not significantly decreased compared with control culture. Ammonia elimination and lidocaine clearance assay showed that the ability of ammonia removal and drug metabolism remained stable.


CONCLUSIONS
Immortalized human hepatocytes can be exposed to AoCLF plasma for at least 24 hours with no significant reduction in the function of cytochrome P450. HepLi-2 cells appear to be effective in metabolism and detoxification and can be potentially used in the development of bioartificial liver.
State Key Laboratory for Diagnosis and Treatment of Infectious Disease, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.
Wei-Bo
WB
Du
Xiao-Ping
XP
Pan
Xiao-Peng
XP
Yu
Cheng-Bo
CB
Yu
Guo-Liang
GL
Lv
Yu
Y
Chen
Lan-Juan
LJ
Li
D000641:Ammonia / Q000378:metabolism; D000779:Anesthetics, Local / Q000493:pharmacokinetics; D001798:Blood Proteins / Q000633:toxicity*; D002461:Cell Line, Transformed; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D058625:End Stage Liver Disease / Q000097:blood* / Q000628:therapy; D005240:Feasibility Studies; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D008658:Inactivation, Metabolic / Q000502:physiology; D008012:Lidocaine / Q000493:pharmacokinetics; D017114:Liver Failure, Acute / Q000097:blood* / Q000628:therapy; D019164:Liver, Artificial
D000779:Anesthetics, Local; D001798:Blood Proteins; D000641:Ammonia; D003577:Cytochrome P-450 Enzyme System; D008012:Lidocaine