The phenolic antioxidant 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA), found in the Pacific oyster Crassostrea gigas, is a superior peroxyl radical scavenger compared to other materials, including Trolox. DHMBA may play an important role in the prevention of health disorders. This study elucidates whether DHMBA prevents the impairment of mineralization of mouse osteoblastic MC3T3-E1 cells under inflammatory conditions by using mouse macrophage RAW264.7 cells in vitro. Culturing with DHMBA (1–100 μM) did not affect the proliferation and death of MC3T3-E1 cells. DHMBA stimulated osteoblastic mineralization. DHMBA blocked the decrease in mineralization of MC3T3-E1 cells caused by culture with the inflammatory cytokine TNF-α. DHMBA inhibited the production of TNF-α by stimulation with lipopolysaccharide (LPS) in RAW264.7 cells. The growth of MC3T3-E1 cells was suppressed by coculture with macrophages under LPS stimulation through the crosstalk of both cells. Interestingly, the growth of MC3T3-E1 cells was suppressed by culturing with the conditioned medium obtained by culturing macrophages with LPS. The effect of the conditioned medium was blocked by the presence of DHMBA or Bay 11–7082, an inhibitor of the TNF-α pathway. The blocking effect of DHMBA was not further enhanced in the presence of Bay 11–7082. Mechanistically, DHMBA was found to decrease the levels of NF-κB p65 and the activity of NF-κB reporter expression in MC3T3-E1 cells. DHMBA was shown to prevent the impairment of osteoblastic mineralization via TNF-α signaling involved in macrophage activation in the bone marrow microenvironment. This study may provide a novel strategy for the therapy of osteoblastic impairment.

在太平洋牡蛎巨牡蛎中发现的酚类抗氧化剂 3,5-二羟基-4-甲氧基苯甲醇 (DHMBA)与其他材料（包括 Trolox）相比，是一种优异的过氧自由基清除剂。DHMBA 可能在预防健康疾病方面发挥重要作用。本研究通过使用小鼠巨噬细胞 RAW264.7 细胞在体外阐明 DHMBA 是否可以防止炎症条件下小鼠成骨细胞 MC3T3-E1 矿化的损害。DHMBA (1–100 μM) 培养不影响 MC3T3-E1 细胞的增殖和死亡。DHMBA 刺激成骨细胞矿化。DHMBA 阻断了炎症细胞因子 TNF-α 培养引起的 MC3T3-E1 细胞矿化减少。DHMBA 通过脂多糖 (LPS) 刺激 RAW264.7 细胞抑制 TNF-α 的产生。在 LPS 刺激下，通过两种细胞的串扰与巨噬细胞共培养，MC3T3-E1 细胞的生长受到抑制。有趣的是，通过用用LPS培养巨噬细胞获得的条件培养基进行培养，MC3T3-E1细胞的生长受到抑制。条件培养基的作用被 DHMBA 或 Bay 11-7082（TNF-α 途径的抑制剂）的存在所阻断。Bay 11-7082 存在时，DHMBA 的阻断效果没有进一步增强。从机制上讲，DHMBA 可降低 MC3T3-E1 细胞中 NF-κB p65 的水平和 NF-κB 报告基因表达的活性。DHMBA 被证明可以通过参与骨髓微环境中巨噬细胞激活的 TNF-α 信号传导来防止成骨细胞矿化受损。这项研究可能为成骨细胞损伤的治疗提供新的策略。