Rhoifolin (ROF) is a main effective component in Citrus grandis ‘Tomentosa’. ROF has a potential anti-inflammatory activity, but its specific effects and mechanisms have not been studied. This study investigated the anti-inflammatory activity of ROF and searched for its possible molecular mechanisms. A mouse model of acute inflammation was induced by lipopolysaccharide, and the effects of ROF on pathological damages of the lung and liver were observed. Carrageenan-induced paw edema rat model was used to evaluate the effect of ROF on the volume of swelling paw. In LPS-induced RAW264.7 macrophages, the expression levels of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α were measured using ELISA. Real-time PCR was used to measure the mRNA levels of iNOS and CCL2. Western blot was used to detect the activation of IκBα and IKKβ in NF-κB signaling pathways. The results showed that ROF accelerated the recoveries of liver and lung tissue damages in acute inflammation mice and inhibited carrageenan-induced paw edema in rats; in addition, ROF significantly suppressed the secretion of TNF-α, IL-1β, and IL-6 in the serum of rats and mouse model. In LPS-induced RAW264.7 cells, 100 μmol/L ROF enhanced cell viability and suppressed the production of TNF-α, IL-6, and IL-1β significantly. ROF also decreased the mRNA expression of iNOS and CCL2 and inhibited IκBα and IKKβ phosphorylation. In summary, ROF had a potential therapeutic value for inflammation. Our research provided experimental basis for the further development of ROF as an anti-inflammatory drug and for clarifying the anti-inflammatory substance basis of Citrus grandis ‘Tomentosa’.

Graphical Abstract

Rhoifolin (ROF) 是柑橘中的主要有效成分'Tomentosa'。ROF具有潜在的抗炎活性，但其具体作用和机制尚未研究。本研究调查了 ROF 的抗炎活性并寻找其可能的分子机制。脂多糖诱导小鼠急性炎症模型，观察ROF对肺和肝脏病理损伤的影响。用角叉菜胶诱导的足部水肿大鼠模型评价 ROF 对肿胀足部体积的影响。在 LPS 诱导的 RAW264.7 巨噬细胞中，使用 ELISA 测量促炎细胞因子 IL-1β、IL-6 和 TNF-α 的表达水平。实时荧光定量 PCR 用于测量 iNOS 和 CCL2 的 mRNA 水平。Western印迹用于检测NF-κB信号通路中IκBα和IKKβ的激活。结果表明，ROF加速了急性炎症小鼠肝脏和肺组织损伤的恢复，并抑制了角叉菜胶引起的大鼠爪水肿；此外，ROF显着抑制大鼠和小鼠模型血清中TNF-α、IL-1β和IL-6的分泌。在 LPS 诱导的 RAW264.7 细胞中，100 μmol/L ROF 可增强细胞活力并显着抑制 TNF-α、IL-6 和 IL-1β 的产生。ROF 还降低 iNOS 和 CCL2 的 mRNA 表达并抑制 IκBα 和 IKKβ 磷酸化。总之，ROF 对炎症具有潜在的治疗价值。我们的研究为ROF作为抗炎药的进一步开发和阐明ROF的抗炎物质基础提供了实验依据。大柑橘'Tomentosa'。

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