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Morphological and biochemical changes in kidney cell lines (RK13, LLC-PK1) treated with ochratoxin A

Ochratoxin A (OTA) is a widespread mycotoxin produced by several species of fungi, Aspergillus, Penicillium and Eurotium. OTA contaminates animal feed and human food. OTA is the potent nephrotoxic, genotoxic and carcinogenic agent, and it affects normal blood coagulation and immune response. It has been implicated as one of the etiologic agents involved in the development of Endemic nephropathy. OTA induces a tubular-interstitial nephropathy in humans and in animals. Toxic effects of OTA in kidneys are usually connected to primary lesions of proximal tubules, followed by spontaneous damage of glomerulus and involution of interstitium. In the present paper we study the effects of OTA on morfologic (cells were stained with hematoxylin and eozin or with phalloidin-FITC labeled and propidium iodid), and biochemical characteristies (intracellular ATP concentration and catalitic concentrations of lactate dehydrogenase, alkaline phosphatase and ƒ×-glutamyltransferase) of RK 13 and LLC PK1 cells in culture. Kidney cell lines were grown at 37oC in RPMI supplemented with 10% fetal calf serum in a humidified atmosphere containing 5% CO2. The cells were then exposed to the different doses of OTA (2, 5 to 25 ƒÝg OTA/ ml of medium) for 24hrs. The number of live cell decreases after treatment with increasing concentration of OTA. 5 ƒÝg OTA/ml reduced the number of leaving cells to 43.4% in the case of RK 13 cells and to 6.2% for LLC PK1 cells, when compared to controls. Results suggest that LLC PK1 cells are more sensitive to OTA than RK 13. The release of LDH into the medium after OTA treatment was much higher in LLC PK1 culture than in RK 13 culture indicating that more intensive necrotic process take places in LLC PK1 culture than in RK 13 culture. OTA treatment affects the intracellular ATP content, but less in RK13 than in LLC PK1. Phalloidin-FITC labelled actin cytoskeleton has been changed in both cell lines.

ochratoxin A; kidney cell lines; RK13; LLC-PK1

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