化学成分分析及鞘柄木（Periploca forrestii Schltr.）促进伤口愈合的潜在活性成分
Chemical profiling and the potential active constituents responsible for wound healing in Periploca forrestii Schltr.
机构信息
Department of Pharmaceutical Analysis, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing 210009, China.
Department of Pharmacology, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing 210009, China.
出版信息
J Ethnopharmacol. 2018 Oct 5;224:230-241. doi: 10.1016/j.jep.2018.04.023. Epub 2018 Apr 20.
ETHNOPHARMACOLOGICAL RELEVANCE
Periploca forrestii Schltr. is a popular folk medicine in china, commonly prescribed for the treatment of rheumatoid arthritis and wounds. The present research aimed to evaluate the effects of HLG on wound healing and reveal the potential active constituents.
MATERIALS AND METHODS
The wound healing activity was assessed by proliferation of fibroblast, migration and collagen production using L929 cells. A reliable HPLC-Q-TOF-MS/MS method was constructed for the systematic identification and characterization of main components in HLG. For further clarifying the potential active ingredients responsible for wound healing, total extract was separated by D101 macroporous resin. The fraction with strongest potency on wound healing was screened out by comparing with total extract. Finally, a new quantitative method was developed for determination of four typical cardiac glycosides in HLG by LC-MS.
RESULTS
The results showed that the total extract significantly promoted proliferation of fibroblast L929 up to 168% at 50 μg/ml. It also notably enhanced L929 migration on day 2 up to 56% and stimulated collagen release (96.1 μg/ml) at 50 μg/ml. A total of 38 compounds were identified or tentatively characterized by HPLC-Q-TOF-MS/MS based on reference substances or literatures. The separation by D101 macroporous adsorption resin led to the identification of 65 ethanol eluate as the most effective fraction. The data suggested that it could markedly promote L929 growth (174% of control), accelerate wound contraction (63% on day 2) and stimulate collagen generation (103.7 μg/ml) at 50 μg/ml, all of which were comparable to those of total extract. Interestingly, the HPLC-Q-TOF-MS/MS analysis revealed that the 65 ethanol fraction was mainly composed of cardiac glycosides. Finally, the new quantitative method was successfully utilized for detection of four typical cardiac glycosides in HLG, showing good performance in terms of analytical methodology.
CONCLUSION
The present study identified the cardiac glycosides as potential active constituents associated with wound healing and might afford a chemical foundation for preparation development of crude drug and quality evaluation of relevant products.
民族药理学相关性
白首乌 Periploca forrestii Schltr. 是中国一种流行的民间药物，常用于治疗类风湿性关节炎和伤口。本研究旨在评估 HLG 对伤口愈合的影响，并揭示其潜在的活性成分。
材料和方法
通过 L929 细胞的增殖、迁移和胶原产生来评估伤口愈合活性。建立了一种可靠的 HPLC-Q-TOF-MS/MS 方法，用于系统鉴定和表征 HLG 中的主要成分。为了进一步阐明负责伤口愈合的潜在活性成分，总提取物通过 D101 大孔树脂进行分离。通过与总提取物比较，筛选出对伤口愈合作用最强的部位。最后，建立了一种新的 LC-MS 定量测定 HLG 中四种典型强心苷的方法。
结果
结果表明，总提取物在 50μg/ml 时可显著促进 L929 成纤维细胞的增殖，达到 168%。它还显著增强了 L929 在第 2 天的迁移，达到 56%，并刺激胶原释放（50μg/ml 时为 96.1μg/ml）。基于对照品或文献，通过 HPLC-Q-TOF-MS/MS 共鉴定或推测出 38 种化合物。通过 D101 大孔吸附树脂分离得到 65 份乙醇洗脱液，为最有效部位。结果表明，它能明显促进 L929 生长（对照的 174%）、加速伤口收缩（第 2 天 63%）和刺激胶原生成（50μg/ml 时为 103.7μg/ml），与总提取物相当。有趣的是，HPLC-Q-TOF-MS/MS 分析表明，65 份乙醇部位主要由强心苷组成。最后，建立了一种新的定量方法，用于检测 HLG 中的四种典型强心苷，该方法在分析方法学方面表现出良好的性能。
结论
本研究鉴定出强心苷是与伤口愈合相关的潜在活性成分，可为原料药的开发和相关产品的质量评价提供化学依据。
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