Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining, Qinghai, China; Qinghai Provincial Key Laboratory of Tibetan Medicine Research, Xining, Qinghai, China; University of Chinese Academy of Science, Beijing, China.
Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining, Qinghai, China; Qinghai Provincial Key Laboratory of Tibetan Medicine Research, Xining, Qinghai, China.
J Ethnopharmacol. 2022 Feb 10;284:114785. doi: 10.1016/j.jep.2021.114785. Epub 2021 Oct 28.
Saussurea obvallata (DC.) Edgew. is a traditional Tibetan medicine used for the treatment of inflammation-related diseases, but the scientific validation was very limited.
This study aimed to rapid screen and targeted isolate cyclooxygenase-2 (COX-2) inhibitors from S. obvallata extract.
An efficient ligand-fishing method based on affinity solid phase extraction (A-SPE) combining with HPLC was developed. The identified COX-2 inhibitors were separated using preparative liquid chromatography. In vitro COX-2 inhibition assays were employed to confirm the inhibitory activities of the isolated compounds. In addition, the effect of the isolated compounds on the production of prostaglandin E2 (PGE2) and the expression of COX-2 in LPS-induced RAW 264.7 were evaluated.
A total of four phenylpropanoids, isolariciresinol, syringaresinol, pinoresinol and balanophonin were targeted isolated as COX-2 inhibitors with IC values of 36.4 ± 2.6 μM, 23.1 ± 1.8 μM, 3.6 ± 0.3 μM and 12.1 ± 0.9 μM, respectively. The isolated compounds significantly inhibited LPS-induced NO production in a dose-dependent manner. And, the results of the inhibitory effect on the release of PGE and the expression of COX-2 in LPS-induced macrophages were consistent with A-SPE analysis.
The present work demonstrated that the developed A-SPE-HPLC method could successfully targeted isolated COX-2 inhibitors from S. obvallata extract. And, the isolation results indicated that the therapeutic effect of S. obvallata on inflammation-related diseases was partly based on the COX-2 active ingredients.
软紫草（DC.）Edgew. 是一种传统的藏药，用于治疗与炎症相关的疾病，但科学验证非常有限。
本研究旨在从软紫草提取物中快速筛选和靶向分离环氧化酶-2（COX-2）抑制剂。
建立了一种基于亲和固相萃取（A-SPE）与 HPLC 相结合的高效配体捕捞方法。采用制备液相色谱法分离鉴定的 COX-2 抑制剂。采用体外 COX-2 抑制试验证实分离化合物的抑制活性。此外，还评估了分离化合物对 LPS 诱导的 RAW 264.7 细胞中前列腺素 E2（PGE2）产生和 COX-2 表达的影响。
共分离得到 4 种苯丙素类化合物，分别为异土木香内酯、丁香树脂酚、松脂醇和巴兰诺菲醇，其 IC50 值分别为 36.4±2.6 μM、23.1±1.8 μM、3.6±0.3 μM 和 12.1±0.9 μM。分离得到的化合物对 LPS 诱导的 NO 产生具有显著的剂量依赖性抑制作用。而且，抑制 LPS 诱导的巨噬细胞中 PGE 释放和 COX-2 表达的作用与 A-SPE 分析结果一致。
本研究表明，所建立的 A-SPE-HPLC 方法可成功地从软紫草提取物中靶向分离 COX-2 抑制剂。分离结果表明，软紫草治疗与炎症相关疾病的疗效部分基于 COX-2 活性成分。
