基于光谱效应关系分析和NF-κB途径的当归dahurica根的抗炎活性成分的筛选

当归Dahurica Root（ADR）是一种通常在中国和其他亚洲国家使用的草药，对各种炎症性疾病具有抗炎作用。但是，尚不清楚导致ADR抗炎作用的生物活性成分和基本机制。这项工作试图发现抗炎的生物活性化合物，并基于频谱效应关系分析和NF-κB信号传导途径探索其在ADR中的基本机制。通过使用二极管阵列检测（HPLC-DAD）的高性能液相色谱（HPLC-DAD）建立ADR样品的色谱指纹，并选择了11个共同峰。然后，采用高性能液相色谱与四极杆的飞行时间质谱法（HPLC-Q/TOF-MS）用于鉴定ADR中的11个公共峰，同时，通过抑制NO，interleukin-1β（Ill-1β），Interlesy-in-el-1β（Inther-irly-1β）评估ADR样品的抗炎活性（TNF-α）在LPS诱导的RAW264.7细胞中产生。研究了ADR样品中HPLC指纹中11个共同峰与抗炎作用之间的频谱效应关系，以通过灰色关系分析（GRA）（GRA）和部分最小二乘回归（PLSR）（PLSR）来鉴定潜在的抗炎生物活性化合物。光谱效应关系分析结果表明，包括Bergapten，Xanthotoxin，phellopterin，Isoimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimimporatin，Xanthotoxolin和Imperatorin在内的六种香豆素化合物可能是ADR中潜在的抗炎生物活性化合物。进一步的验证实验还表明，这六个香豆素在LPS诱导的RAW264.7细胞中表现出对NO，IL-1β，IL-6和TNF-α产生的显着抑制作用。此外，通过评估NF-κB信号传导途径中的蛋白质水平，进行了蛋白质印迹分析以探索两种潜在抗炎生物活性化合物（Phellopterin和Isoimpratorin）的机制。 The western blot results illustrated that phellopterin and isoimperatorin could significantly down-regulate the phosphorylated NF-κB p65 (p-p65), phosphorylated IκBα (p-IκBα) and iNOS, and depress the pro-portion of p-p65/p65 and p-IκBα/IκBα, which indicated that these two coumarins in ADR could potentially exert通过抑制NF-κB途径来抗炎作用。

Angelica dahurica root (ADR), a commonly utilized herbal medicine in China and other Asian nations, which has anti-inflammatory effects on diverse inflammatory ailments. However, the bioactive components and underlying mechanism responsible for the anti-inflammatory effect of ADR are still unclear. This work attempted to discover the anti-inflammatory bioactive compounds and explore their underlying mechanism in ADR based on spectrum-effect relationship analysis and NF-κB signaling pathway. Chromatographic fingerprints of ADR samples were established by high performance liquid chromatography with diode array detection (HPLC-DAD), and a total of eleven common peaks were selected. Then, high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-Q/TOF-MS) was employed for identification of eleven common peaks in ADR Meanwhile, the anti-inflammatory activities of ADR samples were assessed by inhibition of NO, interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) production in LPS-induced RAW264.7 cells. The spectrum-effect relationships between the eleven common peaks in HPLC fingerprints and anti-inflammatory effects of ADR samples were investigated to identify the potential anti-inflammatory bioactive compounds by grey relational analysis (GRA) and partial least squares regression (PLSR). The spectrum-effect relationship analysis results indicated that six coumarin compounds, including bergapten, xanthotoxin, phellopterin, isoimperatorin, xanthotoxol and imperatorin could be potential anti-inflammatory bioactive compounds in ADR. The further validation experiments also showed that these six coumarins demonstrated significant inhibition of NO, IL-1β, IL-6, and TNF-α production in LPS-induced RAW264.7 cells. In addition, western blot analysis was conducted to explore the mechanisms of two potential anti-inflammatory bioactive compounds (phellopterin and isoimperatorin) by assessing the protein levels in the NF-κB signaling pathway. The western blot results illustrated that phellopterin and isoimperatorin could significantly down-regulate the phosphorylated NF-κB p65 (p-p65), phosphorylated IκBα (p-IκBα) and iNOS, and depress the pro-portion of p-p65/p65 and p-IκBα/IκBα, which indicated that these two coumarins in ADR could potentially exert anti-inflammatory effects by suppressing of NF-κB pathway.