Hepatic covalent adduct formation with zomepirac in the CD26-deficient mouse.
OBJECTIVE
Zomepirac (ZP), a non-steroidal anti-inflammatory drug (NSAID), has been reported to cause immune-mediated liver injury. In vivo, ZP is metabolized to a chemically reactive acyl glucuronide conjugate (ZAG) which can undergo covalent adduct formation with proteins. Such acyl glucuronide-derived drug-protein adducts may be important in the development of immune and toxic responses caused by NSAID. We have shown using immunoabsorptions that the 110 kDa CD26 (dipeptidyl peptidase IV) is one of the hepatic target proteins for covalent modification by ZAG. In the present study, a CD26-deficient mouse strain was used to examine protein targets for covalent modification by ZP/metabolites in the liver.


RESULTS
The CD26-deficient phenotype was confirmed by immunohistochemistry, flow cytometry analysis, RT-PCR, enzyme assay and immunoblotting. Moreover, by using monoclonal antibody immunoblots, CD26 was not detected in the livers of ZP-treated CD26-deficient mice. Immunoblots using a polyclonal antiserum to ZP on liver from ZP-treated mice showed three major sizes of protein bands, in the 70, 110 and 140 kDa regions. Most, but not all, of the anti-ZP immunoreactivity in the 110 kDa region was absent from ZP-treated CD26-deficient mice.


CONCLUSIONS
These data definitively showed that CD26 was a component of ZP-modified proteins in vivo. In addition, the data suggested that at least one other protein of approximately 110 kDa was modified by covalent adduct formation with ZAG.
Wang
Min
M
Department of Medicine, University of Queensland, Royal Brisbane Hospital, Brisbane, Australia.
Gorrell
Mark D
MD
Abbott
Catherine A
CA
Jaggi
Rene
R
Marguet
Didier
D
Dickinson
Ronald G
RG
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism* / Q000494:pharmacology; D018819:Dipeptidyl Peptidase 4 / Q000378:metabolism*; D020719:Glucuronides / Q000378:metabolism*; D015151:Immunoblotting; D008099:Liver / Q000201:enzymology; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014046:Tolmetin / Q000031:analogs & derivatives* / Q000378:metabolism* / Q000494:pharmacology
D000894:Anti-Inflammatory Agents, Non-Steroidal; D020719:Glucuronides; C020549:zomepirac; D014046:Tolmetin; D018819:Dipeptidyl Peptidase 4