Background and objective: The novel marine factor 3,5-dihydroxy-4- methoxybenzyl alcohol (DHMBA) was originally identified in the Pacific oyster Crassostrea Gigas. DHMBA has been shown to prevent oxidative stress by scavenging radicals and enhance the production of antioxidant proteins. However, the pharmacologic role of DHMBA has been poorly understood. 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 that cause diverse disease conditions. Therefore, this study has been undertaken to elucidate whether DHMBA expresses anti-inflammatory effects in in vitro mouse macrophage RAW264.7 cells. Methods: Mouse macrophage RAW264.7 cells were cultured in a medium containing 10% fetal bovine serum (FBS) with or without DHMBA (1-1000 μM). Results: Culturing with DHMBA (1-1000 μM) suppressed the growth and stimulated the death of RAW264.7 cells in vitro, leading to a decrease in cell number. Treatment with DHMBA reduced the levels of Ras, PI3K, Akt, MAPK, phospho-MAPK, and mTOR, which are signalling factors to promote cell proliferation, and it raised the levels of p53, p21, Rb, and regucalcin, which are cell growth suppressors. DHMBA treatment elevated caspase-3 and cleaved caspase-3 levels. Interestingly, DHMBA treatment repressed the production of inflammatory cytokines, including tumor necrosis factor-α, interleukin-6, interleukin-1β, or prostaglandin E2, which were enhanced by LPS stimulation. Notably, the levels of NF-κB p65 were increased by LPS treatment, and this augmentation was repres-sed by DHMBA treatment. Moreover, LPS treatment stimulated osteoclastogenesis of RAW264.7 cells. This stimulation was blocked by DHMBA treatment, and this effect was not caused by the presence of an NF-κB signalling inhibitor. Conclusion: DHMBA was found to potentially suppress the activity of inflammatory macrophages in vitro, suggesting its therapeutic usefulness in inflammatory conditions.

中文翻译：

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海洋因子 3,5-二羟基-4-甲氧基苯甲醇抑制体外小鼠巨噬细胞 RAW264.7 细胞中的细胞生长、炎症细胞因子的产生和 NF-κB 信号增强的破骨细胞生成


背景和目的：新型海洋因子 3,5-二羟基-4-甲氧基苯甲醇 (DHMBA) 最初是在太平洋牡蛎巨牡蛎中发现的。 DHMBA 已被证明可以通过清除自由基来预防氧化应激并增强抗氧化蛋白的产生。然而，人们对 DHMBA 的药理作用知之甚少。炎症与许多疾病的发病机制有关。炎症细胞因子在脂多糖 (LPS) 的刺激下在巨噬细胞中产生，并被用作导致多种疾病的生物标志物。因此，本研究旨在阐明 DHMBA 是否在体外小鼠巨噬细胞 RAW264.7 细胞中表达抗炎作用。方法：小鼠巨噬细胞RAW264.7细胞在含10%胎牛血清（FBS）、含或不含DHMBA（1-1000 μM）的培养基中培养。结果：DHMBA (1-1000 μM) 培养抑制体外 RAW264.7 细胞的生长并刺激其死亡，导致细胞数量减少。 DHMBA 治疗降低了 Ras、PI3K、Akt、MAPK、磷酸-MAPK 和 mTOR 的水平，这些信号因子是促进细胞增殖的信号因子，并且它提高了 p53、p21、Rb 和 regucalcin 的水平，这些是细胞生长的信号因子抑制器。 DHMBA 治疗提高了 caspase-3 和裂解的 caspase-3 水平。有趣的是，DHMBA 治疗抑制了炎症细胞因子的产生，包括肿瘤坏死因子-α、白细胞介素-6、白细胞介素-1β 或前列腺素 E2，这些细胞因子通过 LPS 刺激而增强。值得注意的是，LPS 处理增加了 NF-κB p65 的水平，而 DHMBA 处理则抑制了这种增加。此外，LPS 处理刺激 RAW264.7 细胞的破骨细胞生成。 这种刺激被 DHMBA 治疗阻断，并且这种效应不是由 NF-κB 信号传导抑制剂的存在引起的。结论：DHMBA 在体外被发现可能抑制炎症巨噬细胞的活性，表明其在炎症性疾病中的治疗作用。