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Endosulfan‐induced apoptosis and glutathione depletion in human peripheral blood mononuclear cells: Attenuation by N ‐acetylcysteine
Author(s) -
Ahmed Tanzeel,
Tripathi Ashok K.,
Ahmed Rafat S.,
Das Shukla,
Suke Sanvidhan G.,
Pathak Rahul,
Chakraboti Ayanabha,
Banerjee Basu Dev
Publication year - 2008
Publication title -
journal of biochemical and molecular toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.526
H-Index - 58
eISSN - 1099-0461
pISSN - 1095-6670
DOI - 10.1002/jbt.20240
Subject(s) - apoptosis , peripheral blood mononuclear cell , glutathione , oxidative stress , dna fragmentation , programmed cell death , chemistry , endosulfan , microbiology and biotechnology , fragmentation (computing) , acetylcysteine , annexin , biochemistry , biology , in vitro , antioxidant , enzyme , ecology , pesticide , agronomy
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 µM) of endosulfan (α/β at 70:30 mixture) alone and in combination with NAC (20 µM) 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 µM, 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 µM/10 µM 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. © 2008 Wiley Periodicals, Inc. J Biochem Mol Toxicol 22:299–304, 2008; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/jbt.20240