Isoniazid-induced hepatotoxicity in rat hepatocytes of gel entrapment culture.
Gel entrapment culture of rat hepatocytes in hollow fibers were evaluated as a potential in vitro model for studies on isoniazid-induced hepatotoxicity. After exposure to isoniazid (0.11 mM and 1.1 mM) for 24-96 h, gel entrapped hepatocytes were more severely damaged than hepatocyte monolayers according to the assays on methyl thiazolyl tetrazolium (MTT) reduction, intracellular glutathione (GSH) content, reactive oxygen species (ROS) levels, and albumin secretion. Furthermore, CYP 2E1 activity detected by 4-nitrocatechol (4-NC) formation maintained at least 7 days in gel entrapped hepatocytes but decreased to an undetectable level within 2 days in hepatocyte monolayer. And the addition of CYP 2E1 inhibitor, diethyl-dithiocarbamate (DDC), significantly reduced isoniazid-induced GSH depletion in gel entrapped hepatocytes. In addition, the protective effects of N-acetylcysteine (NAC), GSH, liquorice extract and glycyrrhizic acid (GA), a purified compound from liquorice extract, against isoniazid hepatotoxicity were clearly observed in gel entrapped hepatocytes at 72 h incubation. Overall, gel entrapped hepatocytes were more susceptible to isoniazid-induced hepatotoxicity than hepatocyte monolayers by a possible mechanism that higher CYP 2E1 activity in gel entrapped hepatocytes could enhance isoniazid toxicity. This indicates that gel entrapped hepatocytes in hollow fibers could be a more effective model than hepatocyte monolayer for hepatotoxicity research in vitro.
Shen
Chong
C
College of Materials Science and Chemical Engineering, Zhejiang University, Zhejiang, PR China.
Zhang
Hongzi
H
Zhang
Guoliang
G
Meng
Qin
Q
D000111:Acetylcysteine / Q000378:metabolism; D000818:Animals; D000931:Antidotes / Q000494:pharmacology; D000995:Antitubercular Agents / Q000037:antagonists & inhibitors / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury / Q000473:pathology* / Q000517:prevention & control; D019392:Cytochrome P-450 CYP2E1 / Q000378:metabolism; D004050:Ditiocarb / Q000494:pharmacology; D005260:Female; D005978:Glutathione / Q000378:metabolism; D006035:Glycyrrhiza; D019695:Glycyrrhizic Acid / Q000494:pharmacology; D022781:Hepatocytes / Q000187:drug effects*; D007538:Isoniazid / Q000037:antagonists & inhibitors / Q000633:toxicity*; D010936:Plant Extracts / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D012709:Serum Albumin / Q000378:metabolism
D000931:Antidotes; D000995:Antitubercular Agents; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D012709:Serum Albumin; D019695:Glycyrrhizic Acid; D004050:Ditiocarb; D019392:Cytochrome P-450 CYP2E1; D005978:Glutathione; D007538:Isoniazid; D000111:Acetylcysteine