Inactivation of hepatic enzymes by inhalant nitrite—In vivo and in vitro studies

We examined the effects of acute isobutyl nitrite (ISBN) exposure on the activity of several hepatic enzymes. Two strains of adult male mice (Balb/c and C57BL/6) were exposed to 900 ppm ISBN or ambient air for 45 minutes. The enzyme activity of hepatic cytochrome P450 (CYP)-mediated deethylation, glutathione S-transferase (GST), and carboxylesterase (CBE) was monitored through the substrates 3-cyano-7-ethoxycoumarin (CEC), 1-chloro-2,4-dinitrobenzene, and p-nitrophenyl acetate, respectively. Acute ISBN exposure led to a significant reduction in hepatic CYP-mediated CEC deethylation, GST, and CBE activity in Balb/c mice (of 81.5%, 74.7%, and 25.2%, respectively, vs control mice, each at P<.05) when livers were harvested immediately after inhalant exposure. The corresponding decreases in C57BL/6 mice were smaller (with reductions of 21.8%, 18.8%, and 13.3%, respectively, each at P<.05). This enzyme activity, tested in C57BL/6 mice only, returned to control values after a 24-hour period of nonexposure. Follow-up mechanistic investigations using rat liver GST indicated that ISBN-mediated enzyme inactivation was not caused by its metabolites: inorganic nitrite ion (NO2−) or nitric oxide. This inactivation could be prevented, but not reversed, by added glutathione, suggesting irreversible protein oxidation. Using different NO donors as comparative agents, we found that GST inactivation by ISBN was not associated with protein S-nitrosylation or disulfide formation, but with tyrosine nitration. Inhalant nitrite exposure, therefore, led to a significant reduction in hepatic enzyme activity in mice, possibly through tyrosine nitration of hepatic proteins. This effect raises the possibility of drug-drug metabolic interactions from inhalant nitrite abuse. However, determining the applicability of these findings to humans will require further study

Soil bacteria, nitrite and the skin

Little is known about the composition of the skin microbiome and its potential significance for health and disease in the context of the ‘hygiene hypothesis’. We here propose that mammals evolved with a dermal microflora that contributed to the regulation of body physiology by providing nitrite from commensal ammonia-oxidising bacteria in response to ammonia released during sweating. We further hypothesise that modern skin hygiene practices have led to a gradual loss of these bacteria from our skin. Together with other lifestyle-related changes associated with an insufficient bodily supply with nitrite and depletion of other nitric oxide(NO)-related species, a condition we here define as ‘nitropenia’, this has led to a perturbation of cellular redox signalling which manifests as dysregulated immunity and generalised inflammation. If proven correct, this scenario would provide an additional evolutionary rationale and a mechanistic basis for the simultaneous rises in prevalence of a number of seemingly unrelated chronic illnesses over the last 3–4 decades