OBJECTIVE The currently available anti-inflammatory drugs often cause diverse side effects with long-term use. Exploring anti-inflammatory drugs with better efficacy and lower toxicity presents an ongoing challenge. Aloperine is an alkaloid extracted from the leaves and seeds of Sophora alopecuroides L. However, the anti-inflammatory effects of Aloperine have not been fully elucidated. This study aimed to investigate whether Aloperine suppresses lipopolysaccharide (LPS)-induced inflammatory responses in RAW264.7 macrophages. METHODS RAW264.7 macrophages were stimulated with LPS (1 μg/mL) in the presence or absence of Aloperine (50 and 100 μM). mRNA expression was measured by real-time PCR, and protein expression was assessed by western blot analysis. The secretion of pro-inflammatory cytokines was measured by ELISA. The levels of nitric oxide (NO) and reactive oxygen species (ROS) were measured by staining. The transcriptional activity of NF-κB was assayed by a luciferase activity assay. RESULTS The results proved that Aloperine inhibited the expression of LPS-induced pro-inflammatory cytokines [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-17A (IL-17A)] in macrophages. Treatment with Aloperine inhibited NO production through suppressing inducible nitric oxide synthase (iNOS) expression and the secretion of prostaglandin E2 (PGE2) by inhibiting cyclooxygenase 2 (COX-2) expression. Aloperine prevented LPS-induced oxidative stress by reducing the generation of ROS. Furthermore, aloperine significantly reduced Toll-like receptor 4 (TLR4) and myeloid differentiation factor (Myd-88) levels and prevented the nuclear translocation of nuclear factor-κB (NF-κB) in LPS-treated macrophages. CONCLUSION Taken together, our findings show that Aloperine could suppress LPS-induced macrophage activation by inhibiting the TLR4/Myd-88/NF-κB pathway.

中文翻译：

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Aloperine通过抑制TLR4 /NF-κB途径抑制LPS诱导的巨噬细胞活化。

目的目前可使用的消炎药长期使用经常会引起多种副作用。探索具有更好疗效和更低毒性的抗炎药物提出了一个持续的挑战。Aloperine是一种从苦参叶和种子中提取的生物碱。但是，Aloperine的抗炎作用尚未得到充分阐明。这项研究旨在调查Aloperine是否抑制RAW264.7巨噬细胞中脂多糖（LPS）诱导的炎症反应。方法在存在或不存在Aloperine（50和100μM）的情况下，用LPS（1μg/ mL）刺激RAW264.7巨噬细胞。通过实时PCR测量mRNA表达，并通过蛋白质印迹分析评估蛋白表达。通过ELISA测量促炎细胞因子的分泌。通过染色测量一氧化氮（NO）和活性氧（ROS）的水平。通过荧光素酶活性测定法测定NF-κB的转录活性。结果结果证明，Aloperine抑制巨噬细胞中LPS诱导的促炎细胞因子[肿瘤坏死因子-α（TNF-α），白介素-6（IL-6）和白介素-17A（IL-17A）]的表达。 。用阿罗匹林治疗可通过抑制诱导型一氧化氮合酶（iNOS）的表达和通过抑制环氧合酶2（COX-2）的表达而分泌前列腺素E2（PGE2），从而抑制NO的产生。Aloperine通过减少ROS的生成来防止LPS诱导的氧化应激。此外，苦参碱可显着降低Toll样受体4（TLR4）和髓样分化因子（Myd-88）的水平，并防止LPS处理的巨噬细胞中核因子-κB（NF-κB）的核易位。结论综上所述，我们的研究结果表明，Aloperine可以通过抑制TLR4 / Myd-88 /NF-κB途径来抑制LPS诱导的巨噬细胞活化。