A new approach to studying ochratoxin A (OTA)-induced nephrotoxicity: expression profiling in vivo and in vitro employing cDNA microarrays.
Ochratoxin A (OTA) is a mycotoxin often found in cereals as a contaminant, and it is known to cause severe nephrotoxicity in animals and humans. There have been several investigations studying the mode of action of this toxicant, suggesting inhibition of protein synthesis, formation of DNA adducts, and provocation of DNA single-strand breaks as a result of oxidative stress, but little is known about the transcriptional alterations underlying OTA-derived nephrotoxicity so far. We carried out DNA microarray analyses to assess OTA-specific expression profiles in vivo and in vitro. Cultures of primary rat proximal tubular cells and male Wistar rats were treated with a low dose (5 microM and 1 mg/kg, respectively) or a high dose (12.5 microM and 10 mg/kg, respectively) of OTA for 24 or 72 h. Microarray experiments were carried out after dual fluorescent labeling of sample cDNA, and data analysis was performed utilizing different statistical methods. Validity of selected microarray data was confirmed by quantitative real-time PCR. We were able to demonstrate that microarray data derived from our proximal tubule cell (PTC) culture model were highly comparable to the in vivo situation. Marked treatment-specific transcriptional changes were detected for genes involved in DNA damage response and apoptosis (upregulation of GADD 153, GADD 45, annexin V), response to oxidative stress (differential expression of hypoxia-inducible factor 1 and catalase), and inflammatory reactions (upregulation of alpha 2 macroglobulin, ceruloplasmin, and cathepsin S). We conclude that our results provide a molecular basis for interpretation of OTA-induced nephrotoxicity.
Luhe
Anke
A
Department of Molecular and Genetic Toxicology, Bayer AG, Aprather Weg 18a, 42096 Wuppertal, Germany. anke.luehe.al@bayer-ag.de
Hildebrand
Heinz
H
Bach
Ute
U
Dingermann
Theodor
T
Ahr
Hans-Jurgen
HJ
D000210:Acute-Phase Reaction / Q000235:genetics / Q000378:metabolism; D000284:Administration, Oral; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D002273:Carcinogens / Q000008:administration & dosage / Q000633:toxicity*; D018929:Cell Culture Techniques; D004247:DNA / Q000032:analysis; D004249:DNA Damage / Q000187:drug effects / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D004260:DNA Repair / Q000187:drug effects / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D020869:Gene Expression Profiling; D007674:Kidney Diseases / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology; D007687:Kidney Tubules, Proximal / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008297:Male; D009183:Mycotoxins / Q000008:administration & dosage / Q000633:toxicity*; D009793:Ochratoxins / Q000008:administration & dosage / Q000633:toxicity*; D020411:Oligonucleotide Array Sequence Analysis; D018384:Oxidative Stress / Q000187:drug effects / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction
D002273:Carcinogens; D017931:DNA Primers; D009183:Mycotoxins; D009793:Ochratoxins; C025589:ochratoxin A; D004247:DNA