Efficacy of Glutathione in Ameliorating Sulfur Mustard Analog-Induced Toxicity in Cultured Skin Epidermal Cells and in SKH-1 Mouse Skin In Vivo

Abstract

Exposure to chemical warfare agent sulfur mustard (HD) is reported to cause GSH depletion, which plays an important role in HD-linked oxidative stress and skin injury. Using the HD analog 2-chloroethyl ethyl sulfide (CEES), we evaluated the role of GSH and its efficacy in ameliorating CEES-caused skin injury. Using mouse JB6 and human HaCaT epidermal keratinocytes, we observed both protective and therapeutic effects of exogenous GSH (1 or 10 mM) in attenuating a CEES-caused decrease in cell viability and DNA synthesis, as well as S and G 2 M phase arrest in cell cycle progression. However, the protective effect of GSH was stronger than its ability to reverse CEES-induced cytotoxic effect. The observed effect of GSH could be associated with an increase in intracellular GSH levels after its treatment before or after CEES exposure, which strongly depleted cellular GSH levels. N-Acetyl cysteine, a GSH precursor, also showed both protective and therapeutic effects against CEES-caused cytotoxicity. Buthionine sulfoximine, which reduces cellular GSH levels, caused an increased CEES cytotoxicity in both JB6 and HaCaT cells. In further studies translating GSH effects in cell culture, pretreatment of mice with 300 mg/kg GSH via oral gavage 1 h before topical application of CEES resulted in significant protection against CEEScaused increase in skin bifold and epidermal thickness, apoptotic cell death, and myeloperoxidase activity, which could be associated with increased skin GSH levels. Together, these results highlight GSH efficacy in ameliorating CEES-caused skin injury and further support the need for effective antioxidant countermeasures against skin injury by HD exposure.

From the Cover: Catalytic Antioxidant Rescue of Inhaled Sulfur Mustard Toxicity

“…Although AEOL 10150 did not appear to improve GSH levels, this effect may be complicated by the formation of SM-GSH adducts which would not have been detected by the HPLC methods used here (Batal et al, 2015;Mol et al, 2008). Other publications investigating oxidative stress and vesicants have concluded that reductions in the measured markers of oxidative stress are linked to lowered toxicity (Gould et al, 2009;Laskin et al, 2010;O'Neill et al, 2010;Tewari-Singh et al, 2011;Ucar et al, 2007). Our results support the hypothesis that AEOL 10150 lowered oxidative stress in this model, which correlated with attenuating SM toxicity.…”

Protective effects of the thiol compounds GSH and NAC against sulfur mustard toxicity in a human keratinocyte cell line

“…Thiol compounds such as GSH and NAC have been suggested as potential antidotes against SM toxicity in vivo and in vitro (Amir et al, 1998;Laskin et al, 2010;Lindsay et al, 1997;Paromov et al, 2007;Tewari-Singh et al, 2011). Recent data from animal studies in a domestic swine model suggest a beneficial effect of NAC treatment, even when administered after SM exposure (Jugg et al, 2013).…”

“…The presented study focussed on dose-effect relationships with particular interest on cytotoxicity and the release of pro-inflammatory cytokines. Doses of NAC (1, 5 and 10 mM) were established from the available literature on the clinical use of NAC in patients with paracetamol poisoning (Borgstrom et al, 1986;Holdiness, 1991;Shen et al, 2011) and from in vitro studies on the use of NAC against SM toxicity (Atkins et al, 2000;Tewari-Singh et al, 2011). GSH was used at identical molar concentrations.…”

“…Various studies had already been conducted to investigate the potential benefit effects of NAC or GSH treatment before or after SM poisoning (Atkins et al, 2000;Bobb et al, 2005;Rappeneau et al, 2000;Saberi and Zaree, 2009;Tewari-Singh et al, 2011). However, there is limited information regarding dose-effect relationships and, in particular, influence of thiol compound treatment on the secretion of pro-inflammatory cytokines in SMexposed cells.…”

Guidelines for Burn Care Under Austere Conditions

“…38 Other mechanisms include release of reactive oxygen species, depletion of glutathione, generation of reactive nitrogen species by iNOS, and production of proinflammatory cytokines like tumor necrosis factor α. [84][85][86][87] HD is radiomimetic, meaning that it predominantly affects rapidly dividing cells in the GI tract and marrow. Basal keratinocytes are particularly vulnerable to HD, which is why skin injuries feature dermal-epidermal separation (similar histologically to toxic epidermal necrolysis syndrome).…”