Endosulfan‐induced apoptosis and glutathione depletion in human peripheral blood mononuclear cells: Attenuation by N‐acetylcysteine

Abstract

Present study investigated whether endosulfan, an organochlorine pesticide is able to deplete glutathione (GSH) and induce apoptosis in human peripheral blood mononuclear cells (PBMC) in vitro. The role of oxidative stress in the induction of apoptosis was also evaluated by the measurement of the GSH level in cell lysate. The protective role of N-acetylcysteine (NAC) on endosulfan-induced apoptosis was also studied. Isolated human PBMC were exposed to increasing concentrations (0-100 microM) of endosulfan (alpha/beta at 70:30 mixture) alone and in combination with NAC (20 microM) up to 24 h. Apoptotic cell death was determined by Annexin-V Cy3.18 binding and DNA fragmentation assays. Cellular GSH level was measured using dithionitrobenzene. Endosulfan at low concentrations, i.e., 5 and 10 microM, did not cause significant death during 6 h/12 h incubation, whereas a concentration-dependent cell death was observed at 24 h. DNA fragmentation analysis revealed no appreciable difference between control cells and 5 microM/10 microM endosulfan treated cells, where only high molecular weight DNA band was observed. Significant ladder formation was observed at higher concentration, which is indicative of apoptotic cell death. Intracellular GSH levels decreased significantly in endosulfan-treated cells in a dose-dependent manner, showing a close correlation between oxidative stress and degree of apoptosis of PBMC. Cotreatment with NAC attenuated GSH depletion as well as apoptosis. Our results provide experimental evidence of involvement of oxidative stress in endosulfan-mediated apoptosis in human PBMC in vitro.

Molecular Mechanisms of Pesticide Toxicity

“…Coadministration of the antioxidants N-acetylcysteine, which enhances GSH synthesis, attenuated phosphamidon-induced apoptosis. This work supports the hypothesis that oxidative stress, as indicated by GSH depletion, results in the induction of apoptosis by release of cytochrome C (Ahmed et al, 2008;Ahmed et al, 2010). .…”

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

“…On the basis of the Annexin-V-Cy3 binding assay, the cells were categorized as nonapoptotic (green), apoptotic (green and pink), and necrotic (pink) as described by us recently [8].…”

“…The assay was carried out by electrophoresis of isolated DNA in agarose gel as previously documented [8]. Briefly, cultured cells were collected by centrifugation at 1500 rpm for 5 min at 4…”

Clinical Aspects of the Poisoning by the Pesticide Endosulfan

“…57 Endosulfan is also able to deplete glutathione (GSH) and induce apoptosis in human peripheral blood mononuclear cells (PBMC) in vitro. 58 Oral rat administration of 10 mg kg -1 body weight/day for two and four weeks showed toxic interference with liver and kidney´s biochemistry and histology. Biochemical parameters, like aspartate amino transferase, alanine amino transferase, acid phosphatase, alkaline phosphatase, bilirubin, urea and creatinine, were increased, which clearly showed the hepatotoxic and nephrotoxic effects of endosulfan.…”