Effectiveness of Cytoprotective Agents on Sulfur Mustard Induced Toxicity: The In vitro Model
Sulphur mustard (bis (2-chloroethyl) sulfide, SM) is a powerful vesicating chemical warfare agent that causes profound injuries to the eyes, lungs and skin. Despite intensive research following the first use of SM in World War I, there is still no useful pretreatment or therapeutic antidote available. This agent remains a constant chemical threat. A potential approach to combating the debiliting effects of this agent is the use of compounds that can react with this material before it interacts with critical macromolecules. Glutathione (GSH), a tripeptide that exists in high concentrations in cells, reacts with SM and is involved in SM detoxification. Amifostine is a synthetic aminothiol, has been extensively used as a radioprotector. This prompted us to evaluate the protective efficacy of GSH, Amifostine and DRDE-07 (S-2(2- aminoethyl amino) ethyl phenyl sulfide) (synthesized in our lab) against SM toxicity in vitro in HeLa cell line. All these compounds are thiol group containing compounds. Pretreatment of HeLa cell with these cytoprotectants led to decrease in cytotoxicity after SM exposure. The protective efficacy of above compounds were evaluated against sulphur mustard using HeLa cells. The above compounds were added to the media 1 hr before the SM exposure and incubated for 24 hrs. cell viability by MTT assay and LDH leakage were measured as end point.
2. Dacre JC, Goldman M. Toxicology and pharmacology of the chemical warfare agent sulfur mustard. Pharmacol Rev. 1996; 48: 289–326.
3. Gautam A, Gupta A, Lomash V, Pant SC and Vijayaraghavan R. Prophylactic efficacy of combination of DRDE-07 and its analogues with amifostine against sulphur mustard induced sytemic toxicity; Indian journal of Experimental Biology. 2010; 48: 752-761.
4. Gautam A and Vijayaraghavan R. Prophylactic effect of Gossypin against percutaneously administered Sulphur mustards in rats. Biomedical Environ. Science. 2007; 20: 250-259.
5. Pant SC and Lomash V. Sulphur Mustard Induced Toxicity, Mechanism of Action and Current medical management. Defence Life Science Journal. 2016; 1 (1): 07-17.
6. Momeni AZ, Enschaeih S, Meghdadi M. Skin manifestations of mustard gas: A clinical studies of 535 patients exposed to mustard gas. Arch Dermatol. 1992; 128:775-80.
7. Wormser U. Toxicology of mustard gas. Trends Pharmacol. Sci. 1991; 12: 164-67.
8. Smith WJ and Dunn MA. Medical defence against blistering chemical warfare agents. Arch Dermat. 1991; 127:1207-213.
9. Eisenmenger W, Drasch G, Von Clarmann M, Kretshmer E. Clinical and morphological findings on mustard gas [bis(2-chloro ethyl) sulphide] poisoning. J. Forensic Sci. 1991; 36:1688-698.
10. Somani SM and Babu SR. Toxicodynamics of sulphur mustard. Int. J. Clin. Pharmacol. Theor. Toxicol. 1989; 27: 419-435.
11. Arroyo CM, Schafer RJ, Kurt EM, Broomfield CA, Carmichael AJ. Response of normal human keratinocytes to sulfur mustard (SM), cytokine release using a non-enzymatic detachment procedure. Hum. Exp. Toxicol. 1999; 18: 1-11.
12. Kehe K and Szinicz L. Medical aspects of sulphur mustard poisoning. Toxicol. 2005; 214:198-209.
13. Pechura CM and Rall DP. Veterans at Risk: The Health Effects of Mustard Gas and Lewisite. National Academy Press, Washington, DC, p. 428 (1993).
14. Somani SM, Babu SR. Toxicodynamics of sulphur mustard, Int J Clin Pharmacol Ther Toxicol. 1989; 27: 419-35.
15. Papirmeister B, Feister AJ, Robinson SI, Ford RD. Medical Defense against Mustard Gas, Toxic Mechanisms and Pharmacological Implications, CRC Press, Boca Raton, FL (1991).
16. Casillas RP, Kiser RC, Truxell JA, Singer AW, Shumaker SS, Niamuth NA. Therapeutic approaches to dermatotoxicity by sulfur mustard:I Modulation of sulfur mustard induced cutaneous injury in mouse ear vesicant model. J.Appl. Toxicol. 2000; 20: S-145-51.
17. Vijayaraghavan R, Sugendran K, Pant SC, Husain K, Malhotra RC. Dermal intoxication of mice with bis(2-chloroethyl) sulphide and the protective effect of flavonoids. Toxicol. 1991; 69: 35–42.
18. Vojvodic V, Milosavljevic Z, Boskovic B, Bojanic N. The protective effect of different drugs in rats poisoned by sulfur and nitrogen mustards. Fundam. Appl. Toxicol. 1985; 5: S160–S168.
19. Dabrowska MI, Becks LL, Lelli JL, Levee MG, Hinshaw DB. Sulfur mustard induces apoptosis and necrosis in endothelial cells. Toxicol. Appl. Pharmacol. 1996; 141:568-583.
20. Kisby GE, Springer N, Spencer PS. In vitro neurotoxic and DNA-damaging properties of nitrogen mustard. J. Appl. Toxicol. 2000; 20:S35–S41.
21. Vijayaraghavan R, Gautam A, Sharma M, Satish HT, Pant SC, Ganeshan K. Comparative evaluation of some flavonoids and tocopherol acetate against the systemic toxicity induced by sulphur mustard. Indian J Pharmacol. 2009; 40: 114-120.
22. Vijayaraghaven R, Kulkarni A, Kumar P, Lakshmana Rao PV, Pant SC, Pathak U, Raza SK, Jaiswal DK. Prophylactic efficacy of amifostine and DRDE-07 against sulphur mustard administered through various routes. In Pharmacological Perspectives of Some Toxic Chemicals and Antidotes. Flora SJS, Romano JA, Baskin SI, Sekhar K (eds) Narosa Publishers: New Delhi: 25-39 (2001).
23. Gautam A, Vijayaraghavan R. Prophylactic effect of Gossypin against percutaneously administered sulphur mustards in mice.Biomed Environ Sci. 2007; 20: 250-259.
24. Sawyer TW. Modulation of sulphur mustard toxicity by arginine analogues and related nitric oxide synthase inhibitors in vitro. Toxicol. Sci. 1998; 46: 112–121.
25. Sawyer TW. Characterization of protective effects of L-nitroarginine methyl ester (L-NAME) against the toxicity of sulphur mustard in vitro. Toxicol. 1998; 131: 21–32.
26. Gautam A, Vijayaraghavan R, Pant SC, Kumar O, Singh S & Satishkumar HT. Protective effect of quercetin against sulphur mustard-induced oxidative stress in mice. Def. Sci. J. 2007; 57: 707-720.
27. Sharma M, Vijayaraghavan R & Gautam A. DRDE07 and its analogues as promising cytoprotectants to nitrogen mustard (HN-2)—An alkylating anticancer and chemical warfare agent. Toxicol. Lett. 2009; 188: 243-50.
28. Bhattacharya R, Lakshmana Rao PV, Pant SC, Kumar P, Tulsawani RK, Pathak U, Kulkarni A, Vijayaraghavan R. Protective Effects of Amifostine and Its Analogues on Sulfur Mustard Toxicity in Vitro and in Vivo. Toxicol and Appl Pharmacol. 2001; 176: 24–33.
29. Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immun. Methods. 1983; 65: 55-63.
30. Sawyer TW and Risk D. Effects of selected arginine analogues on sulphur mustard toxicity in human and hairless guinea pig skin keratinocytes. Toxicol. Appl. Pharmacol. 2000; 163: 75–85.
31. Sawyer TW, Lundy PM, Weiss MT. Protective effect of an inhibitor of niric oxide synthase on sulphur mustard toxicity in vitro. Toxicology 1996; 141: 138–144.
32. Lindsay CD and Hambrook JL. Protection of A549 cells against the toxic effects of sulphur mustard by hexamethylenetetramine. Hum. Exp. Toxicol. 1997; 16:106–114.
33. Papirmeister B, Gross CL, Meier HL, Petrali JP, Johnson JB. Molecular basis for sulfur mustard-induced vesication. Fundam. Appl. Toxicol. 1985; 5: S134– S149.
34. Ochi T and Miyaura S. Cytotoxicity of an organic hydroperoxide and cellular antioxidant defense system against hydroperoxides in cultured mammalian cells. Toxicol. 1989; 55:69– 82
35. Giuliani I, Baeza-Squiban A, Marano F. Early cytotoxic effects of chlorethamine, a nitrogen mustard, on mammalian airway epithelium. Toxicol. In Vitro. 1997; 11:695–702.
36. Rappeneau S, Baeza-Suiban A, Braut-Boucher F, Aubery M, Gendron MC, Marano F. Use of fluorescent probes to assess the early sulfhydryl depletion and oxidative stress induced by mechlorethamine on airway epithelium. Toxicol. In Vitro 1999; 13: 765-771.
37. Hospers GA, Eisenhauer EA, de Vries E G. The sulfhydryl containing compounds WR-2721 and glutathione as radio and chemoprotectiveagents. A review, indications for use and prospects. Br. J. Cancer. 1999; 80: 629–638