Combined effect of testosterone and apocynin on nitric oxide and superoxide production in PMA-differentiated THP-1 cells

Abstract

Human inducible nitric oxide synthase (iNOS) is most readily observed in macrophages from patients with inflammatory diseases like atherosclerosis. The aim of the present study was to find out the combined effect of male sex hormone; testosterone and apocynin (NADPH oxidase inhibitor) on cytokine-induced iNOS production. THP-1 cells were differentiated into macrophages by phorbol myristate acetate (PMA). Expression of iNOS was induced by the addition of cytokine mixture? Testosterone was added at different concentrations (10 -6-10 -12 M) with apocynin (1 mM). Testosterone (10 -8, 10 -10 M) inhibited NO x production in cytokine-added THP-1 cells which was further confirmed by quantikine assay of iNOS protein and RT-PCR analysis. Testosterone treatment decreased 40% of superoxide anion production. Testosterone showed inhibition of NADPH oxidase, especially expression of p67 phox and p47 phox (cytosol subunits). Addition of testosterone with apocynin further decreased the expression of p67 phox and p47 phox subunits of NADPH oxidase. The findings of the present study suggest that, testosterone; the male androgen plays an important role in the prevention of atherogenesis. Even though apocynin does not have any role on NO production, addition of apocynin together with testosterone is effective in suppressing iNOS activity. © 2004 Elsevier B.V. All rights reserved.