School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing, 210009, China.
State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, 210009, China.
J Nat Med. 2018 Jun;72(3):675-685. doi: 10.1007/s11418-018-1202-1. Epub 2018 Apr 21.
Rheumatoid arthritis (RA) is one of the most prevalent chronic inflammatory and angiogenic diseases. The aim of this study was to evaluate the anti-inflammatory and anti-angiogenic activities in vitro of eight diterpenoids isolated from Daphne genkwa. LC-MS was used to identify diterpenes isolated from D. genkwa. The anti-inflammatory and anti-angiogenic activities of eight diterpenoids were evaluated on LPS-induced macrophage RAW264.7 cells and TNF-α-stimulated human umbilical vein endothelial cells (HUVECs) using hierarchical cluster analysis (HCA) and principal component analysis (PCA). The eight diterpenes isolated from D. genkwa were identified as yuanhuaphnin, isoyuanhuacine, 12-O-(2'E,4'E-decadienoyl)-4-hydroxyphorbol-13-acetyl, yuanhuagine, isoyuanhuadine, yuanhuadine, yuanhuaoate C and yuanhuacine. All the eight diterpenes significantly down-regulated the excessive secretion of TNF-α, IL-6, IL-1β and NO in LPS-induced RAW264.7 macrophages. However, only 12-O-(2'E,4'E-decadienoyl)-4-hydroxyphorbol-13-acetyl markedly reduced production of VEGF, MMP-3, ICAM and VCAM in TNF-α-stimulated HUVECs. HCA obtained 4 clusters, containing 12-O-(2'E,4'E-decadienoyl)-4-hydroxyphorbol-13-acetyl, isoyuanhuacine, isoyuanhuadine and five other compounds. PCA showed that the ranking of diterpenes sorted by efficacy from highest to lowest was 12-O-(2'E,4'E-decadienoyl)-4-hydroxyphorbol-13-acetyl, yuanhuaphnin, isoyuanhuacine, yuanhuacine, yuanhuaoate C, yuanhuagine, isoyuanhuadine, yuanhuadine. In conclusion, eight diterpenes isolated from D. genkwa showed different levels of activity in LPS-induced RAW264.7 cells and TNF-α-stimulated HUVECs. The comprehensive evaluation of activity by HCA and PCA indicated that of the eight diterpenes, 12-O-(2'E,4'E-decadienoyl)-4-hydroxyphorbol-13-acetyl was the best, and can be developed as a new drug for RA therapy.
类风湿关节炎（RA）是最常见的慢性炎症和血管生成疾病之一。本研究旨在评估从芫花中分离得到的八种二萜的体外抗炎和抗血管生成活性。采用 LC-MS 鉴定芫花中二萜的组成。采用层次聚类分析（HCA）和主成分分析（PCA）评价 8 种二萜对 LPS 诱导的 RAW264.7 巨噬细胞和 TNF-α 刺激的人脐静脉内皮细胞（HUVEC）的抗炎和抗血管生成活性。从芫花中分离得到的 8 种二萜分别鉴定为芫花脂素、异芫花脂碱、12-O-（2'E，4'E-癸二烯酰基）-4-羟基-13-乙酰基佛波醇、芫花定、异芫花定碱、芫花定碱、芫花酸 C 和芫花脂碱。所有 8 种二萜均显著下调 LPS 诱导的 RAW264.7 巨噬细胞中 TNF-α、IL-6、IL-1β和 NO 的过度分泌。然而，只有 12-O-（2'E，4'E-癸二烯酰基）-4-羟基-13-乙酰基显著降低 TNF-α 刺激的 HUVEC 中 VEGF、MMP-3、ICAM 和 VCAM 的产生。HCA 得到 4 个聚类，包含 12-O-（2'E，4'E-癸二烯酰基）-4-羟基-13-乙酰基、异芫花脂碱、异芫花定碱和其他 5 种化合物。PCA 表明，按疗效从高到低对二萜进行排序，排名依次为 12-O-（2'E，4'E-癸二烯酰基）-4-羟基-13-乙酰基、芫花脂素、异芫花脂碱、芫花脂碱、芫花酸 C、芫花定、异芫花定碱、芫花定碱。总之，从芫花中分离得到的 8 种二萜在 LPS 诱导的 RAW264.7 细胞和 TNF-α 刺激的 HUVEC 中表现出不同程度的活性。HCA 和 PCA 的综合活性评价表明，8 种二萜中，12-O-（2'E，4'E-癸二烯酰基）-4-羟基-13-乙酰基活性最好，可开发为治疗 RA 的新药。
