The marine factor 3,5-dihydroxy-4-methoxybenzyl alcohol expresses the anti-inflammatory effects in mouse macrophages RAW264.7 cells in vitro

Abstract

Background: Inflammation is implicated in the pathogenesis of many diseases. Inflammatory cytokines are produced in macrophages with stimulation of lipopolysaccharide (LPS) and are used as biomarkers participating in diverse disease conditions. The novel marine factor 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA) was initially identified in the Pacific oyster Crassostrea Gigas. DHMBA has properties to reduce oxidative stress as radical scavenging and increase the production of antioxidant proteins. The pharmacologic role of DHMBA, however, has been poorly understood.Methods and Results: This study has been undertaken to investigate whether DHMBA attenuates growth, cytokine production, and osteoclastogenesis in inflammatory mouse macrophage RAW264.7 cells. Culturing with DHMBA (1-1000 µM) suppressed the growth and stimulated the death of RAW264.7 cells in vitro, leading to decrease in cell number. Mechanistically, DHMBA treatment decreased the levels of Ras, PI3K, Akt, MAPK, phospho-MAPK, and mTOR of signaling factors to promote the proliferation, and it raised the levels of p53, p21, Rb, and regucalcin, which are cell growth suppressors. The levels of caspase-3 and cleaved caspase-3 were increased by DHMBA treatment. Culturing with DHMBA suppressed productions of inflammatory cytokines, including tumor necrosis factor-α, interleukin-6, interleukin-1β, or prostaglandin E2, were enhanced by LPS stimulation. Notably, the levels of NF-κB p65 were increased by LPS treatment, and this increase was repressed by DHMBA treatment. LPS treatment stimulated osteoclastogenesis of RAW264.7 cells. This stimulation was blocked by DHMBA treatment.Conclusion: DHMBA was found to potentially suppress the activity of inflammatory macrophages in vitro, suggesting therapeutic usefulness in inflammatory conditions.