Citrus hystrix DC. (CH) is found in many countries in Southeast Asia. This plant has been reported for anti-microbial, anti-cancer and anti-inflammatory bioactivities. However, the anti-inflammatory and anti-inflammasome properties of the leaves remain poorly understood. This study aimed to investigate the effect of CH leaves on NLRP3 and NF-κB signaling pathways. CH leaves were sequentially extracted using hexane, ethyl acetate and 95% ethanol to give three crude extracts. An active compound, lupeol was fractionated from the ethanolic extract using chromatographic techniques, and its structure was identified and confirmed by spectroscopic methods. Anti-inflammatory activities were observed on both lipopolysaccharide-stimulated and NLRP3 adenosine triphosphate-induced macrophages. The release of pro-inflammatory cytokines (IL-1β, IL-6 and TNF-α) was analyzed by Enzyme-Linked Immunosorbent Assay (ELISA). Real-time qRT-polymerase chain reaction (PCR) was used to measure inflammatory-associated gene expression. NF-κB protein expressions were investigated using the immunoblotting technique. The active fraction of ethanolic CH leaves and lupeol significantly reduced the release of pro-inflammatory cytokines and suppressed the expression of both inflammasome genes and NF-κB proteins. The ethanolic extract of CH leaves and lupeol showed potent anti-inflammatory activities by targeting NF-κB and NLRP3 signaling pathways.

中文翻译：

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柑桔DC的植物化学成分。叶通过巨噬细胞中的NF-κB信号和NLRP3炎症小体活性减弱炎症

柑橘DC。（CH）在东南亚的许多国家中都可以找到。据报道该植物具有抗微生物，抗癌和抗炎的生物活性。然而，人们对树叶的抗炎和消炎特性仍然知之甚少。这项研究旨在调查CH叶对NLRP3和NF-κB信号通路的影响。依次使用己烷，乙酸乙酯和95％乙醇萃取CH叶片，得到三种粗萃取物。使用色谱技术从乙醇提取物中分离出一种活性化合物羽扇豆酚，并通过光谱法鉴定和确认了其结构。在脂多糖刺激和NLRP3腺苷三磷酸诱导的巨噬细胞上均观察到抗炎活性。促炎细胞因子（IL-1β，通过酶联免疫吸附测定（ELISA）分析IL-6和TNF-α）。实时qRT-聚合酶链反应（PCR）用于测量炎症相关基因的表达。使用免疫印迹技术研究了NF-κB蛋白的表达。乙醇性CH叶片和羽扇豆酚的活性级分显着降低了促炎细胞因子的释放，并抑制了炎性体基因和NF-κB蛋白的表达。CH叶和羽扇豆酚的乙醇提取物通过靶向NF-κB和NLRP3信号通路显示出有效的抗炎活性。乙醇性CH叶片和羽扇豆酚的活性级分显着降低了促炎细胞因子的释放，并抑制了炎性体基因和NF-κB蛋白的表达。CH叶和羽扇豆酚的乙醇提取物通过靶向NF-κB和NLRP3信号通路显示出有效的抗炎活性。乙醇性CH叶片和羽扇豆酚的活性级分显着降低了促炎细胞因子的释放，并抑制了炎性体基因和NF-κB蛋白的表达。CH叶和羽扇豆酚的乙醇提取物通过靶向NF-κB和NLRP3信号通路显示出有效的抗炎活性。