The role of oxidative stress in the ochratoxin A-mediated toxicity in proximal tubular cells.
Balkan endemic nephropathy (BEN), a disease characterized by progressive renal fibrosis in human patients, has been associated with exposure to ochratoxin A (OTA). This mycotoxin is a frequent contaminant of human and animal food products, and is toxic to all animal species tested. OTA predominantly affects the kidney and is known to accumulate in the proximal tubule (PT). The induction of oxidative stress is implicated in the toxicity of this mycotoxin. In the present study, primary rat PT cells and LLC-PK(1) cells, which express characteristics of the PT, were used to investigate the OTA-mediated oxidative stress response. OTA exposure of these cells resulted in a concentration-dependent elevation of reactive oxygen species (ROS) levels, depletion of cellular glutathione (GSH) levels and an increase in the formation of 8-oxoguanine. The OTA-induced ROS response was significantly reduced following treatment with alpha-tocopherol (TOCO). However, this chain-braking anti-oxidant did not reduce the cytotoxicity of OTA and was unable to prevent the depletion of total GSH levels in OTA-exposed cells. In contrast, pre-incubation of the cell with N-acetyl-L-cysteine (NAC) completely prevented the OTA-induced increase in ROS levels as well as the formation of 8-oxoguanine and completely protected against the cytotoxicity of OTA. In addition, NAC treatment also limited the GSH depletion in OTA-exposed PT- and LLC-PK(1) cells. From these data, we conclude that oxidative stress contributes to the tubular toxicity of OTA. Subsequently, cellular GSH levels play a pivotal role in limiting the short-term toxicity of this mycotoxin in renal tubular cells.
Schaaf
G J
GJ
Department of Veterinary Pharmacology, Pharmacy and Toxicology (VFFT) Faculty of Veterinary Medicine, Utrecht University, P.O. Box 80152, NL 3508 TD Utrecht, The Netherlands.
Nijmeijer
S M
SM
Maas
R F M
RF
Roestenberg
P
P
de Groene
E M
EM
Fink-Gremmels
J
J
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D005978:Glutathione / Q000032:analysis / Q000378:metabolism; D006147:Guanine / Q000031:analogs & derivatives* / Q000032:analysis / Q000096:biosynthesis; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000378:metabolism; D009183:Mycotoxins / Q000633:toxicity*; D009793:Ochratoxins / Q000633:toxicity*; D018384:Oxidative Stress; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis; D051381:Rats; D017382:Reactive Oxygen Species / Q000032:analysis / Q000378:metabolism; D013997:Time Factors
D000975:Antioxidants; D009183:Mycotoxins; D009793:Ochratoxins; D011506:Proteins; D017382:Reactive Oxygen Species; C025589:ochratoxin A; C024829:8-hydroxyguanine; D006147:Guanine; D005978:Glutathione