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Assessment of phosphamidon-induced apoptosis in human peripheral blood mononuclear cells: Protective effects of N-acetylcysteine and curcumin

✍ Scribed by Tanzeel Ahmed; Ashok K. Tripathi; Rafat S. Ahmed; Basu Dev Banerjee

Publisher
John Wiley and Sons
Year
2010
Tongue
English
Weight
221 KB
Volume
24
Category
Article
ISSN
1095-6670

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✦ Synopsis

Abstract

The molecular mechanism for noncholinergic toxicity of phosphamidon, an extensively used organophosphate pesticide, is still not clear. The aim of the present study is to find the possible molecular mechanism of this pesticide to induce apoptosis and the role of different drugs for attenuation of such effects. Human peripheral blood mononuclear cells (PBMC) were incubated with increasing concentrations of phosphamidon (0–20 μM) for 6–24 h. The MTT assay reveals that phosphamidon induces cytotoxicity in a dose‐dependent manner. Cellular glutathione (GSH) is depleted in a dose‐dependent manner from 55% to 70% at concentrations between 10 and 20 μM. The percentage of cells that bind to Annexin‐V, which is a representative of cells either undergoing apoptosis or necrosis during 24 h incubation, increases in a dose‐dependent manner. Above 5 μM, significant necrosis of cells was observed. DNA fragmentation assay revealed that at low concentration of phosphamidon (1 μM), no appreciable change in DNA fragmentation was seen; however, distinct fragmentation was observed beyond 2.5 μM. Phosphamidon was found to cause significant depletion of GSH, which correlates well with the percentage of cells undergoing apoptosis. An increasing trend in levels of cytochrome c was observed with increasing concentration of phosphamidon, indicating that the apoptotic effect of phosphamidon is mediated through cytochrome c release. Coadministration of the antioxidants N‐acetylcysteine and curcumin attenuated phosphamidon‐induced apoptosis. This further supports our hypothesis that oxidative stress, as indicated by GSH depletion, results in the induction of apoptosis by release of cytochrome c. © 2010 Wiley Periodicals, Inc. J Biochem Mol Toxicol 24:286–292, 2010; View this article online at wileyonlinelibrary.com. DOI 10.1002/jbt.20337

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