[Bile salt induces apoptosis of hepatocytes: the mechanism of hepatic function injury during obstructive jaundice].
OBJECTIVE
To determine if the bile salt induces hepatocyte apoptosis in vitro.


METHODS
Hepatocytes were isolated by in situ collagenase perfusion and plated in 6 well flat bottom with DMEM/F12, 0.5 micro/ml insulin. Two hours later, glycochenodeoxcholate (GCDC) 25, 50, 100, 200, 300 micromol was added and cells evaluated by DNA-PI staining FACS and terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) 3, 6, 9, 12, 18, 20, 24 hours later. Agarose gel electrophoresis of DNA extracted from hepatocytes after incubation with GCDC using various experimental conditions.


RESULTS
The hepatocytes treated with GCDC have a high apoptotic rate as compared with the controls. 100 micromol GCDC treated 24 hours, nearly 61.81% of hepatocytes were apoptotic by FACS evaluation. The TUNEL showed that apoptotic hepatocytes had less cell volume and stained with brown by Biotin-11-dUTP. DNA ladder of bile-salt treated hepatocytes were shown in agarose gel electrophoresis.


CONCLUSIONS
The mechanism of hepatic injury during obstructive jaundice is related to bile salt caused hepatocyte apoptosis.
Li
D
D
Depatment of General Surgery, Xuanwu Hospital, Capital University of Medical Sciences, Beijing 100053.
Sun
J
J
Sun
H
H
Li
F
F
Liu
F
F
Liu
S
S
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001647:Bile Acids and Salts / Q000494:pharmacology*; D002779:Cholestasis / Q000503:physiopathology; D004305:Dose-Response Relationship, Drug; D005260:Female; D005999:Glycochenodeoxycholic Acid / Q000494:pharmacology; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D020287:In Situ Nick-End Labeling; D008099:Liver / Q000473:pathology / Q000503:physiopathology; D051381:Rats; D017207:Rats, Sprague-Dawley
D001647:Bile Acids and Salts; D005999:Glycochenodeoxycholic Acid