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Ochratoxin a Induced Apoptosis in Kidley Cell Lines

Ochratoxin A (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. MDCK, LLC-PK1 and RK 13 kidney cell lines were exposed to the different doses of OTA (2.5 to 25  g OTA/mL of medium) for 24 hrs. We studied the effects of OTA on morphology of the cells stained with haematoxylin and eosin or labelled with phalloidin-FITC, TUNEL technique and stained with propidium iodide. We also followed the particular biochemical characteristics, intracellular ATP concentration and catalytic concentration of LDH. The number of living cells significantly decreased after treatment with increasing concentration of OTA. 2.5  g OTA/mL reduced the number of leaving cells to 69.4% for RK 13 cells, to 34.5% for LLC-PK1 and to 27.5% for MDCK cells, when compared to untreated control cells. In MDCK, LLC-PK1 and RK 13 cell lines we have observed 12%, 11% and 8.2%, apoptotic cells, respectively. Results suggest that MDCK and LLC-PK1 cell lines are more sensitive to OTA than RK 13 cells. The release of LDH into the medium after treatment with OTA was more abundant in MDCK and LLC-PK1 than in RK 13 cells indicating more intensive necrotic process. OTA reduced the intracellular ATP content, but less in RK 13 than in MDCK and LLC-PK1 cell lines. Taken all together, OTA induced either apoptotic or necrotic cell death in cell type- and dose- dependent manner.

ochratoxin A; apoptosis; cytoskeleton; intracellular ATP; Endemic nephropathy

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