Use of BCL2 Monoclonal Antibodies to Prove the Prooncogenic and Nephrotoxic Action of Ochratoxine a in Broiler Chickens

摘要

BCL2 antiapoptotic proteins plays a central role in preserving mitochondrial structureand function, preventing the occurrence of transition permeabiliy and inhibiting mitochondrialcytochrome C release in cytosol.The goal of this study was to evaluate the nephrotoxic and prooncogenic effect ofOTA in broiler chickens experimentally intoxicated, using BCL2 antibodies. Experiment wereused 40 Ross 307 broilers six days old, divided into 2 groups: experimental (E) and control(C), kept in the same environmental condition. E group received daily by gavage ochratoxinA (OTA-Sigma Chemicals Co.) eluted in sterilized sunflower oil, at a dose of 50.62 mcg / kgbw and the control group received only eluent (sterilized sunflower oil), for 20 days. At theend of each week during the experiment, five chicks were selected by random from eachgroup and killed. Histopathology was performed on kidney fragments, embedded in paraffin,and stained by HEA method. Immunohistochemistry (IHC) were performed with anti BCL2and DAB Substrate Kit (BD Biosciences Pharmigen). Electronomicroscopic investigationswere carried out with electron microscope TESLA BS 500.In two of the five chicks exposed to OTA for 20 days BCL2 positive cells wereidentified, ordered in layers into the renal papilla, fact considered a presumably indicator ofepithelial cell tumors with transitional cells of the upper urinary tract. The incidence of theseurothelial tumors of renal pelvis and urethra is higher in people in endemic areas forochratoxicosis (Vutelic and Sostaric1991). BCL2 positive cells were found in both proximaland distal convoluted tubules, both detached from the basal membrane but also byneighboring cells, or placed in the lumen of its layered. Some positive BCL2 cells have highnucleus, sometimes two nuclei, rich in euchromatine. BCL2 proteins are known asprooncogenic, regulating the apoptosis. BCL2 overexpression play a role in tumorogenesisand may explain the relative resistance of renal cell carcinoma to chemotherapeutic agentsand radiation therapy.Ultrastructurally, in the kidney, the presence of abnormal forms of mitochondria werenoted. These are numerous, large, oval shapes, spherical granules of different sizeselectronodense other degenerate, characterized by small cristae reduced number of spaces inthe downtown area.