来自Astilbe Grandis的C-14羧基的丁烷三萜类化合物通过抑制NF-κB信号传导途径，抑制了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
影响因子:4.80000
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
摘要
过度的炎症会给人类的身心健康带来重大风险。传统的MIAO医学Astilbe Grandis以其抗炎特性而闻名。但是，该植物中许多化合物的特定抗炎作用和机制尚不清楚。这项研究旨在研究两种特征性的三萜特征性的抗炎作用和机制，即3α-乙酰氧基-12-EN-27-二酸（1）和3β-乙酰氧基-12-en-27- oic酸（2），从努利伯氏症和脂肪popolymacachamageymacachamaChamage（Lipopolymage and France） - lpshamm cancemape（2）分离出来。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的机理。使用Griess方法评估一氧化氮（NO）水平。通过酶联免疫吸收测定法（ELISA）测量了肿瘤坏死因子-Alpha（TNF-α），白介素-6（IL-6）和白介素-1BETA（IL-1β）的浓度。使用蛋白质印迹和定量实时PCR（QRT-PCR）确定环氧合酶-2（COX-2）和诱导型一氧化氮合酶（INOS）的表达。另外，通过蛋白质印迹评估了核因子-KAPPA B（NF-κB）中p65的磷酸化水平。通过免疫荧光染色评估了NF-κBp65的核易位。最后，通过Molecular docking.com验证了化合物与NF-κBp65靶标的结合亲和力显着抑制NO，TNF-α，IL-6，IL-6，IL-1β，COX，COX-2和INOS在LPS诱导的巨噬细胞中的表达。从机械上讲，他们通过下调p65的磷酸化水平和核易位来减弱NF-κB信号传导途径的激活。这项研究阐明了抗炎活性和特征性灰丁烷三萜的特征性机制，并通过C-14羧基的1和2构图通过c-14羧基，通过lps诱导的插条构图，通过抑制了nf-κB。 时间。这些发现表明，这两种化合物将来有望成为抗炎干预措施的潜在候选者。
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.