Benzodiazepines inhibit neutrophil chemotaxis and superoxide production in a stimulus dependent manner; PK-11195 antagonizes these effects

Abstract

Diazepam, which binds both central (neuronal) and peripheral (non-neuronal) benzodiazepine binding sites, and Ro5-4864, a ligand selective for benzodiazepine peripheral binding sites (PBS), both inhibited the FMLP induced chemotaxis in human neutrophils at concentrations as low as 10⁻⁸ M. A selective peripheral benzodiazepine antagonists, PK-11195 (10⁻⁵ M), partially reversed the benzodiazepine inhibition of chemotaxis. Diazepam also inhibited the superoxide production induced by FMLP, NaF, and A23187, but not that induced by PMA whose stimulant action was insensitive even to 10⁻⁴ M diazepam. The FMLP-induced superoxide production was most sensitive to diazepam inhibition (ID₅₀ = 2.25 × 10⁻⁶ M diazepam); the effect of NaF was slightly less sensitive (ID₅₀ = 1.34 × 10⁻⁵ M diazepam); and the effect of A23187 was least sensitive as it was suppressed only at 10⁻⁴ M diazepam concentrations. Like diazepam, Ro5-4864 inhibited the FMLP-induced superoxide production, and PK-11195 (10⁻⁵ M) significantly antagonized both diazepam and Ro5-4864 inhibition.

Binding studies showed the presence of a saturable benzodiazepine ‘peripheral’ type binding site (PBS) on human neutrophils with a $\text{K}{}_{\mathrm{<mtext>d</mtext>}}\text{of}\text{1.2 ± 0.06 × 10}{}^{\mathrm{-8}}\text{M}\text{(±}\text{SEM}\text{),}\text{and a B}{}_{\max }\text{of}\text{1028 ± 86.2}\text{fmol}\text{/10}{}^6\text{Cells}\text{(±}\text{SEM}\text{)}\text{for}\text{[}{}^3\text{H}\text{]}\text{Ro}\text{5-4864}$; the binding was displaceable by PK-11195, Ro5-4864 and diazepam but not by clonazepam.