Coumarin derivatives, such as esculetin, have various physiological functions, including antioxidant, anti-inflammatory, antibacterial, antiviral, and anti-cancer. 6,7-Dihydroxy-4-methylcoumarin (6,7-DH-4MC) is a derivative of esculetin, and its anti-inflammatory effect and mechanism in macrophages have not been studied. In this study, the anti-inflammatory activity of 6,7-DH-4MC was evaluated by measuring the expression of inflammatory factors (NO and PGE₂) and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) in LPS-stimulated RAW 264.7 macrophages. The results revealed that 6,7-DH-4MC significantly reduced NO levels and PGE₂ expression without inducing cytotoxicity; it was confirmed that the inhibition of NO and PGE₂ expression was related to iNOS and COX-2 downregulation in response to 6,7-DH-4MC treatment. Moreover, 6,7-DH-4MC decreased the levels of pro-inflammatory cytokines, such as IL-1β and IL-6, in a dose-dependent manner. Mechanistic studies revealed reduced phosphorylation of ERK and p38-MAPK upon 6,7-DH-4MC treatment. Furthermore, the degradation of IκB-α and phosphorylation of NF-κB in cells treated with LPS were interrupted by 6,7-DH-4MC treatment. These results suggest that 6,7-DH-4MC is a potential therapeutic agent for inflammatory diseases. To the best of our knowledge, this is the first report demonstrating the anti-inflammatory effects of 6,7-DH-4MC in RAW 264.7 cells via MAPK and NF-κB signaling pathways.

香豆素衍生物，如七叶苷，具有多种生理功能，包括抗氧化、抗炎、抗菌、抗病毒和抗癌。6,7-Dihydroxy-4-methylcoumarin (6,7-DH-4MC) 是七叶亭的衍生物，其在巨噬细胞中的抗炎作用和机制尚未研究。在本研究中，通过测量炎症因子（NO 和 PGE ₂）和促炎细胞因子（IL-1β、IL-6 和 TNF-α）的表达来评估 6,7-DH-4MC 的抗炎活性。) 在 LPS 刺激的 RAW 264.7 巨噬细胞中。结果表明，6,7-DH-4MC显着降低NO水平和PGE ₂表达，且不引起细胞毒性；证实了 NO 和 PGE ₂的抑制作用表达与响应 6,7-DH-4MC 处理的 iNOS 和 COX-2 下调有关。此外，6,7-DH-4MC 以剂量依赖性方式降低促炎细胞因子的水平，如 IL-1β 和 IL-6。机制研究表明 6,7-DH-4MC 处理后 ERK 和 p38-MAPK 的磷酸化降低。此外，LPS 处理的细胞中 IκB-α 的降解和 NF-κB 的磷酸化被 6,7-DH-4MC 处理中断。这些结果表明 6,7-DH-4MC 是一种潜在的炎症性疾病治疗剂。据我们所知，这是第一份通过 MAPK 和 NF-κB 信号通路证明 6,7-DH-4MC 在 RAW 264.7 细胞中的抗炎作用的报告。