过度炎症对人类身心健康构成重大风险。落新妇是一种传统的苗药，以其抗炎特性而闻名。然而，这种植物中许多化合物的具体抗炎作用和机制仍不清楚。本研究旨在研究两种特征齐墩果烷三萜类化合物 3α-acetoxyolean-12-en-27-oic Acid (1) 和 3β-acetoxyolean-12-en-27-oic Acid (2) 的抗炎作用和机制，使用脂多糖（LPS）诱导的巨噬细胞从落新妇中分离得到。通过在RAW 264.7细胞和THP-1细胞中建立LPS诱导的炎症模型来研究化合物1和2的抗炎作用和机制。使用格里斯法评估一氧化氮（NO）水平。通过酶联免疫吸附测定（ELISA）测量肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和白细胞介素-1β（IL-1β）的浓度。使用蛋白质印迹和定量实时 PCR (qRT-PCR) 测定环氧合酶-2 (COX-2) 和诱导型一氧化氮合酶 (iNOS) 的表达。此外，还通过蛋白质印迹评估了核因子-κB (NF-κB) 中 p65 的磷酸化水平。通过免疫荧光染色评估 NF-κB p65 的核转位。最后通过分子对接验证了化合物与NF-κB p65靶点的结合亲和力。化合物1和2显着抑制了NO、TNF-α、IL-6、IL-1β、COX-2和iNOS的表达。 LPS诱导的巨噬细胞。从机制上讲，它们通过下调 p65 的磷酸化水平和核转位来减弱 NF-κB 信号通路的激活。本研究阐明了具有 C-14 羧基的特征齐墩果烷三萜化合物 1 和 1 的抗炎活性和潜在机制。 2、首次在LPS诱导的巨噬细胞中通过抑制NF-κB信号通路。这些研究结果表明，这两种化合物有望成为未来抗炎干预的潜在候选者。版权所有 © 2024 Yue、Luo、Zhao、Zhao、Ye、He 和 Zou。

Excessive inflammation poses significant risks to human physical and mental health. Astilbe grandis, a traditional Miao medicine, is renowned for its anti-inflammatory properties. However, the specific anti-inflammatory effects and mechanisms of many compounds within this plant remain unclear. This study aims to investigate the anti-inflammatory effects and mechanisms of two characteristic oleanane triterpenoids, 3α-acetoxyolean-12-en-27-oic acid (1) and 3β-acetoxyolean-12-en-27-oic acid (2), isolated from Astilbe grandis, using lipopolysaccharide (LPS)-induced Macrophages.The anti-inflammatory effects and mechanisms of compounds 1 and 2 were investigated by establishing an LPS-induced inflammation model in RAW 264.7 cells and THP-1 cells. Nitric oxide (NO) levels were assessed using the Griess method. The concentrations of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1beta (IL-1β) were measured via enzyme-linked immunosorbent assay (ELISA). The expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was determined using western blotting and quantitative real-time PCR (qRT-PCR). Additionally, the phosphorylation level of p65 in nuclear factor-kappa B (NF-κB) was assessed through western blotting. The nuclear translocation of NF-κB p65 was assessed through immunofluorescence staining. Finally, the binding affinity of the compounds to NF-κB p65 target was validated through molecular docking.Compounds 1 and 2 significantly inhibited the expression of NO, TNF-α, IL-6, IL-1β, COX-2, and iNOS in LPS-induced Macrophages. Mechanistically, they attenuated the activation of the NF-κB signaling pathway by downregulating the phosphorylation level and nuclear translocation of p65.This study elucidates the anti-inflammatory activities and potential mechanism of the characteristic oleanane triterpenoids with C-14 carboxyl group, compounds 1 and 2, in LPS-induced Macrophages by inhibiting the NF-κB signaling pathway for the first time. These findings suggest that these two compounds hold promise as potential candidates for anti-inflammatory interventions in the future.Copyright © 2024 Yue, Luo, Zhao, Zhao, Ye, He and Zou.