通脉养心丸中心肌保护活性物质的筛选

目的:基于细胞模型以及高效液相色谱—质谱联用技术对通脉养心丸中心肌保护活性物质进行筛选研究。方法:利用高效液相制备技术对通脉养心丸进行制备分离,然后采用过氧化氢氧化损伤H9c2大鼠心肌细胞模型对通脉养心丸活性组分进行筛选。利用高效液相色谱—质谱联用技术对活性组分进行分析,通过查阅文献和根据化合物裂解规律,鉴定推断出可能的活性化合物。结果:发现通脉养心丸中10个组分具有明显的心肌保护活性,考察了其中5个组分的半数有效浓度并发现其呈现良好的量效关系。从这5个组分中推断出11个可能的活性化合物,包含甘草酸、甘草香豆素、甘草利酮、Glyasperin A等。结论:筛选出通脉养心丸中心肌保护活性物质,有望为中药复方制剂活性物质筛选研究提供借鉴。
Abstract: Objective: Based on cell model and HPLC-MS technology, to screen myocardial protection active compounds from traditional patent medicine Tongmai Yangxin pill (TMYXP). Methods: Fractions of TMYXP were prepared by high performance liquid preparation technology. The cardioprotective effects of prepared fractions were tested on H2O2 oxidation-damaged H9c2 myocardiocytes. The active components were analyzed by high performance liquid chromatography (HPLC) coupled with high resolution mass spectrometry. The possible active compounds were putatively identified by comparison of their MS ions and molecular weight with literatures. Results: Ten TMYXP components presented significant myocardial protective activities, 5 of which were investigated and presented good dose-effect relationships. Their median effective concentrations (EC50) were respectively 11.66, 17.44, 13.10, 7.332, 15.15 μg/mL. Totally, 11 potential active compounds were analyzed and identified, including Glycyrrhizic acid, Glycycoumarin, Licoisoflavone, Ophiopogonin D', Licoricon, Gancaonin L, Neoglycyrol, Emodin, Angeloylgomisin H, Angeloylgomisin Q and Glyasperin A.Conclusion: The myocardial protection active compounds of TMYXP were screened successfully. Key words: Myocytes, cardiac Anoxia Chromatography, high pressure liquid Glycyrrhiza uralensis/drug effects Flavones/pharmacology Glycyrrhizic acid/pharmacology Schisandra chinensis Ophiopogon japonicus Disease models, animal