Immunochemical analysis of quinol-thioether-derived covalent protein adducts in rodent species sensitive and resistant to quinol-thioether-mediated nephrotoxicity.
2,3,5-Tris(glutathion-S-yl)hydroquinone (TGHQ) is nephrotoxic in male Fischer 344 rats (20 micromol/kg) and albino guinea pigs (200 micromol/kg), but not BALB/c or B6C3F1 mice or Golden Syrian hamsters (200 micromol/kg). Since quinones are known to alkylate proteins, and because such macromolecular damage may play a role in cytotoxicity, we investigated the covalent binding of TGHQ to kidney (target tissue) and liver (nontarget tissue) of rodents "sensitive" or "resistant" to the nephrotoxic effects of TGHQ. Immunohistochemical staining of tissue obtained 2 h after administration of TGHQ, with rabbit anti-2-bromo-N-(acetyl-L-cystein-S-yl)HQ antibodies, correlated with the subsequent region of necrosis observed 19 h after dosing in Fischer 344 rats and guinea pigs. Immunohistochemical staining was localized to the S3 segment of the renal proximal tubules, at the corticomedullary junction along the medullary rays, and in the outer stripe of the outer medulla. Immunostaining was also observed in the same region in hamsters, but subsequent necrosis did not develop. In contrast, no immunostaining was observed in mice. Moreover, immunostaining was not detected in the livers of any species. Western blot analysis revealed numerous immunoreactive renal proteins in TGHQ-treated animals. The most distinctive immunostaining renal proteins were observed in Fischer 344 rats at approximately 34 kDa (mitochondria), approximately 35 kDa (nuclei) which comigrated with histone H1, and approximately 73 kDa (urine) which comigrated with gamma-glutamyl transpeptidase. These adducted proteins were not detected in other species. Qualitative differences in alkylated proteins may therefore contribute to species susceptibility to TGHQ.
Kleiner
H E
HE
Division of Pharmacology and Toxicology, College of Pharmacy, The University of Texas at Austin, Austin, Texas 78712, USA. hkleiner@sprd1.mdacc.tmc.edu
Jones
T W
TW
Monks
T J
TJ
Lau
S S
SS
D000818:Animals; D006224:Cricetinae; D003600:Cytosol / Q000378:metabolism; D005978:Glutathione / Q000031:analogs & derivatives* / Q000737:chemistry / Q000633:toxicity; D006873:Hydroquinones / Q000737:chemistry* / Q000633:toxicity*; D007150:Immunohistochemistry; D007668:Kidney / Q000201:enzymology / Q000378:metabolism; D007674:Kidney Diseases / Q000139:chemically induced* / Q000473:pathology; D008099:Liver / Q000378:metabolism; D008297:Male; D008647:Mesocricetus; D051379:Mice; D008815:Mice, Inbred Strains; D008861:Microsomes / Q000378:metabolism; D011506:Proteins / Q000737:chemistry*; D011817:Rabbits; D051381:Rats; D011916:Rats, Inbred F344; D013440:Sulfides / Q000737:chemistry* / Q000633:toxicity*
D006873:Hydroquinones; D011506:Proteins; D013440:Sulfides; C078765:2,3,5-(triglutathion-S-yl)hydroquinone; D005978:Glutathione