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Type: Artigo de periódico
Title: Cytoskeletal Rearrangement And Cell Death Induced By Bothrops Alternatus Snake Venom In Cultured Madin-darby Canine Kidney Cells.
Author: Nascimento, Juliana Minardi
Franchi, Gilberto Carlos
Nowill, Alexandre Eduardo
Collares-Buzato, Carla Beatriz
Hyslop, Stephen
Abstract: Bothrops snake venoms cause renal damage, with renal failure being the main cause of death in humans bitten by these snakes. In this work, we investigated the cytoskeletal rearrangement and cytotoxicity caused by Bothrops alternatus venom in cultured Madin-Darby canine kidney (MDCK) cells. Incubation with venom (10 and 100 microg/mL) significantly (p <0.05) decreased the cellular uptake of neutral red dye after 1 and 3 h. Venom (100 microg/mL) also markedly decreased the transepithelial electrical resistance (RT) across MDCK monolayers. Staining with rhodamine-conjugated phalloidin revealed disarray of the cytoskeleton that involved the stress fibers at the basal cell surface and focal adhesion-associated F-actin in the cell-matrix contact region. Feulgen staining showed a significant decrease in the number of cells undergoing mitosis and an increase in the frequency of altered nuclei. Scanning electron microscopy revealed a decrease in the number of microvilli and the presence of cells with a fusiform format. Flow cytometry with annexin V and propidium iodide showed that cell death occurred by necrosis, with little apoptosis, a conclusion supported by the lack of DNA fragmentation characteristic of apoptosis. Pretreating the cells with catalase significantly attenuated the venom-induced loss of viability, indicating a possible involvement of H2O2 in the cellular damage; less protection was observed with superoxide dismutase or N omega-nitro-L-arginine methyl ester. These results indicate that Bothrops alternatus venom is cytotoxic to cultured MDCK cells, possibly via the action of reactive oxygen species. This cytotoxicity could contribute to nephrotoxicity after envenoming by this species.
Subject: Animals
Cell Death
Cell Line
Cell Survival
Crotalid Venoms
Flow Cytometry
Microscopy, Electron, Scanning
Time Factors
Citation: Biochemistry And Cell Biology = Biochimie Et Biologie Cellulaire. v. 85, n. 5, p. 591-605, 2007-Oct.
Rights: fechado
Identifier DOI: 10.1139/o07-067
Date Issue: 2007
Appears in Collections:Unicamp - Artigos e Outros Documentos

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