具有C-14羧基的Oleanane三萜类化合物通过抑制NF-κB信号通路抑制Astilbe grandis来源的Lipopolysaccharide (LPS)诱导的巨噬细胞激活
Oleanane triterpenoids with C-14 carboxyl group from Astilbe grandis inhibited LPS-induced macrophages activation by suppressing the NF-κB signaling pathway
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影响因子:4.8
分区:医学3区 / 药学3区
发表日期:2024
作者:
Lan Yue, Jinfang Luo, Chenliang Zhao, Jinfeng Zhao, Jianghai Ye, Kang He, Juan Zou
DOI:
10.3389/fphar.2024.1413876
摘要
过度的炎症对人体的身心健康构成重大风险。Astilbe grandis,作为一种传统的苗族药材,以其抗炎特性而闻名。然而,该植物中许多化合物的具体抗炎作用及其机制尚不清楚。本研究旨在利用脂多糖(LPS)诱导的巨噬细胞模型,探究从Astilbe grandis中分离的两种特征性Oleanane三萜类化合物,3α-乙酰氧基-olean-12-en-27-羧酸(1)和3β-乙酰氧基-olean-12-en-27-羧酸(2)的抗炎作用及机制。通过建立LPS诱导的RAW 264.7细胞和THP-1细胞炎症模型,评估化合物的抗炎效果。采用Griess法检测一氧化氮(NO)水平,酶联免疫吸附试验(ELISA)测定肿瘤坏死因子α(TNF-α)、白细胞介素-6(IL-6)和白细胞介素-1β(IL-1β)的浓度。采用Western blot和实时定量PCR(qRT-PCR)检测环氧酶-2(COX-2)和诱导型一氧化氮合酶(iNOS)的表达。此外,Western blot还用于检测核因子κB(NF-κB)p65的磷酸化水平,免疫荧光染色用于观察NF-κB p65的核转位。最后,通过分子对接验证化合物与NF-κB p65靶点的结合亲和力。化合物1和2显著抑制LPS诱导的巨噬细胞中NO、TNF-α、IL-6、IL-1β、COX-2及iNOS的表达。从机制上看,它们通过降低p65的磷酸化水平和核转位,减弱NF-κB信号通路的激活。本研究首次阐明了具有C-14羧基的特征性Oleanane三萜类化合物(化合物1和2)在LPS诱导的巨噬细胞中的抗炎活性及潜在机制,主要通过抑制NF-κB信号通路。这些发现提示这两种化合物在未来具有作为抗炎干预候选药物的潜力。
Abstract
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.