S-allyl cysteine in combination with clotrimazole downregulates Fas induced apoptotic events in erythrocytes of mice exposed to lead |
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Authors: | Samir MandalSudip Mukherjee Kaustav Dutta ChowdhuryAvik Sarkar Kankana BasuSoumosish Paul Debasish KarmakarMahasweta Chatterjee Tuli BiswasGobinda Chandra Sadhukhan Gargi Sen |
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Affiliation: | a Indian Institute of Chemical Biology (CSIR), 4, Raja S.C. Mullick Road, Kolkata-700032, Indiab Department of Zoology, Vidyasagar College, 39 Sankar Ghosh Lane, Kolkata-700006, India |
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Abstract: | BackgroundChronic lead (Pb2 +) exposure leads to the reduced lifespan of erythrocytes. Oxidative stress and K+ loss accelerate Fas translocation into lipid raft microdomains inducing Fas mediated death signaling in these erythrocytes. Pathophysiological-based therapeutic strategies to combat against erythrocyte death were evaluated using garlic-derived organosulfur compounds like diallyl disulfide (DADS), S allyl cysteine (SAC) and imidazole based Gardos channel inhibitor clotrimazole (CLT).MethodsMorphological alterations in erythrocytes were evaluated using scanning electron microscopy. Events associated with erythrocyte death were evaluated using radio labeled probes, flow cytometry and activity gel assay. Mass spectrometry was used for detection of GSH-4-hydroxy-trans-2-nonenal (HNE) adducts. Fas redistribution into the lipid rafts was studied using immunoblotting technique and confocal microscopy.ResultsCombination of SAC and CLT was better than DADS and CLT combination and monotherapy with these agents in prolonging the survival of erythrocytes during chronic Pb2 + exposure. Combination therapy with SAC and CLT prevented redistribution of Fas into the lipid rafts of the plasma membrane and downregulated Fas-dependent death events in erythrocytes of mice exposed to Pb2 +.Conclusion and general significanceCeramide generation was a critical component of Fas receptor-induced apoptosis, since inhibition of acid sphingomyelinase (aSMase) interfered with Fas-induced apoptosis during Pb2 + exposure. Combination therapy with SAC and CLT downregulated apoptotic events in erythrocytes by antagonizing oxidative stress and Gardos channel that led to suppression of ceramide-initiated Fas aggregation in lipid rafts. Hence, combination therapy with SAC and CLT may be a potential therapeutic option for enhancing the lifespan of erythrocytes during Pb2 + toxicity. |
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Keywords: | FITC, fluorescein isothiocyanate PS, phosphatidylserine Prx2, peroxiredoxin2 NAD, nicotinamide adenine dinucleotide NADP, nicotinamide adenine dinucleotide phosphate NADH, nicotinamide adenine dinucleotide reduced NADPH, nicotinamide adenine dinucleotide phosphate reduced HPF, 3&prime -(p-hydroxyphenyl) fluorescein FACS, fluorescence-activated cell sorting Pb2 +, lead SAC, S allyl cysteine CLT, clotrimazole ROS, reactive oxygen species OH 0" alt=" radical dot" src=" http://cdn.els-cdn.com/sd/entities/rad" class=" glyphImg" >&minus , hydroxyl radical GSH, glutathione K+, potassium ion DMSA, meso-2, 3-dimercaptosuccinic acid MiADMSA, monoisoamyl meso-2, 3-dimercaptosuccinic acid DADS, diallyl disulfide GST, glutathione S transferase SGPT, serum glutamate pyruvate transaminase SGOT, serum oxaloacetate transaminase WBCs, white blood cells RBCs, red blood cells PBS, phosphate buffer saline HNE, hydroxynonenal TBARS, thiobarbituric acid reactive substance TBA, thiobarbituric acid DTNB, 5,5&prime -dithiobis-2-nitrobenzoic acid CDNB, 1-cloro-2,4-dinitrobenzene FCS, fetal calf serum aSMase, acid sphingomyelinase BSA, bovine serum albumin DMTU, dimethyl thiourea DAS, diallyl sulfide LDL,, low-density lipoprotein &minus SH group, sulfhydryl group H2O2, hydrogen peroxide GPx, glutathione peroxidase DISC, death inducing signaling complex |
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