Investigation of proteomic biomarkers in in vivo hepatotoxicity study of rat liver: toxicity differentiation in hepatotoxicants.
We investigated the overall protein expression profiles in the in vivo hepatotoxicity of rats induced by four well-recognized hepatotoxicants. Acetaminophen (APAP), amiodarone (AMD), tetracycline (TC) and carbon tetrachloride (CTC) were administered to male rats by gavages and the liver at 24 hr post-dosing was applied to the proteomic experiment. Blood biochemistry and histopathology were examined to identify specific changes related to the compounds given. Protein expression in the liver was investigated by 2-dimensional gel electrophoresis (2DE), and spots showing a significantly different expression in treated versus control group were excised from gels and identified by Q-Tof mass spectrometer. They were well characterized based on their functions related to the mechanisms of toxicity of the compounds. Among them, we focused on the 8 proteins that were affected by all 4 compounds examined. Proteins related to oxidative stress response such as carbonic anhydrase III (CA3) and 60kDa heat shock protein (HSP60), and energy metabolism such as adenylate kinase 4 (AK4) were found. Moreover, hierarchical clustering analysis using 2D-gel spots information revealed the possibility to differentiate the groups based on their toxicity levels such as severity of liver damage. These results suggested that assessing the effects of hepatotoxicants on protein expression is worth trying to screen candidate compounds at the developmental stage of drugs.
Worldwide Safety Sciences, Pfizer Global Research & Development, Nagoya Laboratories, Pfizer Inc., 5-2 Taketoyo, Aichi 470-2393, Japan. Toshinori.Yamamoto@pfizer.com
Toshinori
T
Yamamoto
Rie
R
Kikkawa
Hiroshi
H
Yamada
Ikuo
I
Horii
D000082:Acetaminophen / Q000633:toxicity*; D000638:Amiodarone / Q000633:toxicity*; D000818:Animals; D015415:Biomarkers; D002251:Carbon Tetrachloride / Q000633:toxicity*; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D011506:Proteins / Q000378:metabolism*; D040901:Proteomics; D051381:Rats; D011919:Rats, Inbred Strains; D013752:Tetracycline / Q000633:toxicity*
D015415:Biomarkers; D011506:Proteins; D000082:Acetaminophen; D002251:Carbon Tetrachloride; D013752:Tetracycline; D000638:Amiodarone