Lack of a role for creatine phosphate kinase in sulphur mustard-induced cytotoxicity.
Several compounds involved in the creatine phosphate kinase (CPK) pathway were evaluated for their protective effects against the chemical warfare (CW) agent sulphur mustard (HD), in primary chick embryo neuron and first passage human skin keratinocyte cultures. High concentrations of both creatine and creatine phosphate were found to be protective under all culture conditions and increased the LC(50) of HD in both culture systems up to approximately 250%. Little difference was observed in the protective activity of these compounds in undifferentiated versus differentiated neuronal culture, or in proliferating versus differentiating cultures of keratinocytes. The protective effect of these compounds was found to be strictly prophylactic in nature. Although a modest decline in HD half-life was measured in buffer containing creatine phosphate, this did not account for the protective effects of this compound. In contrast to historical literature reporting 90-100% HD-induced CPK inhibition of purified enzyme, less than 30% of CPK activity was found to be inhibited by HD in both human keratinocytes and in swine blood plasma. Incubation of keratinocyte cultures with creatine or creatine phosphate prior to HD exposure did not alter CPK activity, compared with HD-only treated cultures. Although high mM concentrations of both creatine and creatine phosphate exert significant protective effects against HD, these results do not support a role for CPK in its toxicity or in the development of medical countermeasures against this CW agent.
Casualty Management Section, Defence Research and Development Canada-Suffield Box 4000, Medicine Hat, Alberta, Canada T1A 8K6.
P
P
Nelson
A
A
Burczyk
T W
TW
Sawyer
D000818:Animals; D002478:Cells, Cultured; D002619:Chemical Warfare Agents / Q000633:toxicity*; D018890:Chemoprevention; D002642:Chick Embryo; D003401:Creatine / Q000494:pharmacology; D003402:Creatine Kinase / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D015603:Keratinocytes / Q000187:drug effects* / Q000201:enzymology; D009151:Mustard Gas / Q000633:toxicity*; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology; D018696:Neuroprotective Agents / Q000494:pharmacology; D010725:Phosphocreatine / Q000494:pharmacology
D002619:Chemical Warfare Agents; D018696:Neuroprotective Agents; D010725:Phosphocreatine; D003402:Creatine Kinase; D003401:Creatine; D009151:Mustard Gas