College of Pharmacy, Institute of Pharmaceutical Science and Technology, Hanyang University, Ansan 15588, Korea.
Department of Pharmacognosy, Faculty of Pharmacy, University of Sindh, Jamshoro 76080, Pakistan.
Molecules. 2021 Aug 30;26(17):5269. doi: 10.3390/molecules26175269.
In this study, a centrifugal partition chromatography (CPC) separation was applied to identify antioxidant-responsive element (ARE) induction molecules from the crude extract of roots. CPC was operated with a two-phase solvent system composed of -hexane-methanol-water (10:8.5:1.5, //) in dual mode (descending to ascending), which provided a high recovery rate (>95.5%) with high resolution. Then, ARE induction activity of obtained CPC fractions was examined in ARE-transfected HepG2 cells according to the weight ratios of the obtained fractions. The fraction exhibiting ARE-inducing activity was further purified by preparative HPLC that led to isolation of two eudesmane type sesquiterpenes as active compounds. The chemical structures were elucidated as linderolide U () and a new sesquiterpene named as linderolide V () by spectroscopic data. Further bioactivity test demonstrated that compounds and enhanced ARE activity by 22.4-fold and 7.6-fold, respectively, at 100 μM concentration while 5 μM of sulforaphane induced ARE activity 24.8-fold compared to the control.
在这项研究中，采用了一种离心分配色谱（CPC）分离方法，从根的粗提取物中鉴定出抗氧化反应元件（ARE）诱导分子。CPC 采用由正己烷-甲醇-水（10:8.5:1.5，//）组成的两相溶剂系统在双模式（下降到上升）下操作，提供了高回收率（>95.5%）和高分辨率。然后，根据获得的馏分的重量比，在 ARE 转染的 HepG2 细胞中检查获得的 CPC 馏分的 ARE 诱导活性。表现出 ARE 诱导活性的馏分通过制备型 HPLC 进一步纯化，导致两种倍半萜类化合物作为活性化合物分离。通过光谱数据阐明了化合物的化学结构为 linderolide U（）和一种新的倍半萜命名为 linderolide V（）。进一步的生物活性测试表明，化合物和在 100 μM 浓度下分别增强了 ARE 活性 22.4 倍和 7.6 倍，而 5 μM 的萝卜硫素与对照相比，诱导 ARE 活性 24.8 倍。
