Shi Yu, Lei Yueyue, Guo Shiwen, Li Lujun, Li Xu, Liu Xinyi, Ding Shuiping
National & Local Joint Engineering Research Center of High-throughput Drug Screening Technology, State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Key Laboratory of Biotechnology of Chinese Traditional Medicine, College of Health Science and Engineering, Hubei University, Wuhan, China.
Gansu Herbal Medicine Planting Co., Ltd, Lanzhou, China.
Phytochem Anal. 2025 Jun;36(4):943-956. doi: 10.1002/pca.3479. Epub 2024 Nov 12.
Flavonoids are the main nonvolatile component responsible for the anti-rheumatoid arthritis (RA) synovitis activities of Artemisia argyi Folium. However, the effective ingredient remains unidentified. Spectrum-effect relationships analysis was a reliable and efficient strategy for herbal effective ingredient discovery.
This study aimed to screen the effective ingredients within the total flavonoid from Artemisia argyi Folium (TFAA) that exhibit anti-RA synovitis activities based on spectrum-effect relationship.
TFAA was obtained through ethanol extraction and subsequent purification with D101 resin from 15 distinct batches of Artemisia argyi Folium. The fingerprint of TFAA was established using HPLC, and its efficacy against RA synovitis was evaluated by determining the inhibition rate of nitric oxide (NO) on MH7A synovioblast induced by TNF-α. Common peaks were identified using HPLC-MS/MS and authentic standards. The spectrum-effect relationships between the fingerprints and efficacy were analyzed by gray relational analysis (GRA), canonical correlation analysis (CCA), and partial least squares regression analysis (PLSR) to pinpoint the peaks responsible for the anti-RA synovitis activity, and the results were further verified by in vitro anti-RA synovitis experiments and molecular docking studies.
The fingerprint revealed 14 common peaks, and 12 compounds were identified, including four caffeoylquinic acids and eight flavonoids. Among them, five flavonoids-X10 (hispidulin), X11 (jaceosidin), X12 (centaureidin), X13 (eupatilin), and X14 (casticin)-were highly relevant to anti-RA synovitis activity. Verification experiments confirmed their inhibitory effect on NO production and cytokine secretion in MH7A cells, showing anti-RA synovitis potential, which was consistent with the established spectrum effect relationship. The underlying mechanism might be related to the inhibition of iNOS.
Hispidulin, jaceosidin, centaureidin, eupatilin, and casticin were the key effective ingredient of TFAA responsible for its anti-RA synovitis effects. These compounds can serve as quality control markers for Artemisia argyi Folium and as lead compounds for anti-RA synovitis treatment.
黄酮类化合物是艾叶抗类风湿性关节炎（RA）滑膜炎活性的主要非挥发性成分。然而，有效成分仍未明确。谱效关系分析是发现草药有效成分的可靠且高效的策略。
本研究旨在基于谱效关系筛选艾叶总黄酮（TFAA）中具有抗RA滑膜炎活性的有效成分。
通过乙醇提取及随后用D101树脂从15个不同批次的艾叶中纯化得到TFAA。采用高效液相色谱法建立TFAA指纹图谱，并通过测定一氧化氮（NO）对TNF-α诱导的MH7A滑膜成纤维细胞的抑制率来评估其对RA滑膜炎的疗效。使用高效液相色谱-质谱/质谱联用仪和对照品鉴定共有峰。通过灰色关联分析（GRA）、典型相关分析（CCA）和偏最小二乘回归分析（PLSR）分析指纹图谱与疗效之间的谱效关系，以确定具有抗RA滑膜炎活性的峰，并通过体外抗RA滑膜炎实验和分子对接研究进一步验证结果。
指纹图谱显示有14个共有峰，鉴定出12种化合物，包括4种咖啡酰奎宁酸和8种黄酮类化合物。其中，5种黄酮类化合物——X10（滨蓟黄素）、X11（硫磺菊素）、X12（矢车菊素）、X13（泽兰黄素）和X14（紫铆素）——与抗RA滑膜炎活性高度相关。验证实验证实了它们对MH7A细胞中NO产生和细胞因子分泌的抑制作用，显示出抗RA滑膜炎的潜力，这与建立的谱效关系一致。潜在机制可能与抑制诱导型一氧化氮合酶（iNOS）有关。
滨蓟黄素、硫磺菊素、矢车菊素、泽兰黄素和紫铆素是TFAA中具有抗RA滑膜炎作用的关键有效成分。这些化合物可作为艾叶的质量控制标志物以及抗RA滑膜炎治疗的先导化合物。
