Effect of all-trans-retinoic acid on cytokine production in a murine macrophage cell line.

All-trans-retinoic acid (RA) is a cancer chemopreventive agent and a pluripotent morphogen. It belongs to the class of retinoids that, besides being inducers of differentiation and growth-inhibitos, exert immunomodulatory and anti-inflammatory functions by mechanisms that are not clearly understood. Macrophages play different roles in diverse physiological processes, including ones in orchestrating immune and inflammatory responses. Products of activated macrophages such as interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNFalpha), interleukin-6, interleukin-8, and nitric oxide (NO) are important regulators of inflammatory reactions. In this study J774A. 1 cells, a murine macrophage cell line, was used to study the effects of RA on the production of NO, TNFalpha and IL-1beta. Cells were stimulated with lipopolysaccharide (LPS) with or without RA. RA depressed the levels of NO in a dose-dependent manner. NO production and subsequent nitrite accumulation in the media peaked at 24 h, plateaued at 48 h, and remained at the same level through 72 h. The presence of RA decreased TNFalpha levels, measured by both bioassay and enzyme-linked immunosorbent assay (ELISA), but these did not correlate with increased mRNA expression measured by reverse-transcriptase polymerase chain reaction at 6 h after LPS stimulation. IL-1beta protein production measured by both ELISA and bioassay decreased with RA treatment. IL-1beta mRNA expression was not affected by RA except at low doses. This study indicated that RA modulates cytokine production in J774A.1 macrophage cells. Inhibition of inflammatory cytokine production may play a role in the anti-inflammatory activity of RA. The results suggested that effects of RA are complex and are time and concentration dependent.