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DPPH-HPLC-MS assisted rapid identification of endothelial protective substances from Xiao-Ke-An

作者 Yang, Zhenzhong; Li, Zhenhao; Li, Zheng

期刊 JOURNAL OF ETHNOPHARMACOLOGY , JAN 30 2018,:211

DOI 10.1016/j.jep.2017.07.016 PubMed 28716572

摘要 Ethnopharmacological relevance: Xiao-Ke-An formula (XKA) is a Chinese medicine widely used for treating diabetes and associated complications. Endothelial protection is thought to be one of its therapeutic mechanisms. However, the protective effect of XKA on endothelial cells remains unclear, especially in oxidative injury induced endothelial dysfunction.Materials and Methods: A novel strategy to rapidly screen and identify potential endothelial protective substances from XKA was established by combining the 2,2''-diphenyl-l-picrylhydrazyl high performance liquid chromatography coupled with mass spectrometry (DPPH-HPLC-MS) approach with cell-based verification. Firstly, the DPPH-HPLC-MS approach was employed to identify the antioxidants in XKA. Then, the potent endothelial protective effect of XKA, and the potential active substances and mechanism of action were revealed in EA.hy926 cells injured by tert-butyl hydroperoxide (t-BHP).Results: XKA exhibited potent antioxidant activity and endothelial protective effect. Phenolic acids derived from Salvia miltiorrhiza Bunge, root and rhizome, xanthones from Anemarrhena asphodeloides Bunge, rhizome, and acteoside from Rehmannia glutinosa (Gaertn.) DC., root, were identified as the major endothelial protective components in XKA.Conclusions: An efficient method for rapid identifying endothelial protective substances from complex mixtures was developed and used to identify the major endothelial protective components in XKA. This method would help reveal the material base of herbal medicine with endothelial protective effect, and could also be applied to discover novel natural-origin endothelial protective substances.

涉及物种 丹参 Salvia miltiorrhiza 知母 Anemarrhena asphodeloides 地黄 Rehmannia glutinosa