Diisopropylphosphorofluoridate-induced cholinergic hyperactivity and lipid peroxidation

In the present study, the association between acetylcholine (ACh)-induced muscle necrosis and the appearance of lipid peroxidation products was investigated. Lipid peroxidation in this injury was quantified by the malondialdehyde-thiobarbituric acid complex (TBA-MDA) using HPLC. To induce muscle necrosis, rats were treated with 1.0 or 2.0 mg/kg diisopropylphosphorofluoridate (DFP), an irreversible inhibitor of AChE that induced muscle fasciculations, and were euthanized 30-120 min after the DFP treatment. DFP caused a dose-dependent increase in AChE inhibition, muscle fasciculations, TBA-MDA formation, and muscle necrosis. Reduction of glutathione (GSH) by pretreatment with buthionine sulfoximine (BSO) potentiated the DFP-induced changes in TBA-MDA and caused an increase in the number of necrotic muscle fibers. Prevention of fasciculations by pretreatment with cholinergic antagonists such as atropine and d-tubocurarine, before DFP, inhibited the increase in lipid peroxidation, and significantly attenuated the muscle fiber necrosis. Without affecting muscle fasciculations, the antioxidant U-78517F prevented the increase in lipid peroxidation and reduced the number of muscle fibers that became necrotic. It is suggested that DFP-induced AChE inhibition causes pronounced muscle hyperactivity as the initial step that triggers free radical-induced lipid peroxidation as the final common pathway to muscle injury.