逍遥散对皮质酮诱导的PC12损伤模型保护作用及神经保护成分分析验证研究

Study on the protective effect of Xiaoyaosan on corticosterone-induced PC12 injury model and its neuroprotective components

摘要:

目的 通过探究逍遥散的神经保护作用及其化学组成，发现逍遥散中可能发挥神经保护作用的有效活性成分。方法 采用水提法提取逍遥散，通过高分辨质谱-液相色谱质谱联用仪分析提取液的化学成分；制备逍遥散含药血清；构建皮质酮（Corticosterone，CORT，400 μmol/L）诱导PC12细胞损伤模型，检测含药血清对PC12细胞增殖率的影响；用SIMCA软件进行细胞增殖率-成分含量相关性分析，找到逍遥散中可能具有神经保护作用的化合物。结果 与模型组相比，逍遥散含药血清可减轻CORT诱导的细胞损伤，并促进细胞增殖（P=0.000）。液质谱图结合软件分析比对发现，主要药效化合物有：芍药苷、甘草酸、甘草苷、迷迭香酸、没食子酸、2-苯基乙醛、（15Z）-9，12，13-Trihydroxy-15-octadecenoic acid、丹酚酸B、甘草皂苷G2等，单体验证结果显示苯乙醛具有一定的神经保护作用（P=0.000）。结论 证明逍遥散具有神经保护作用，分析并鉴定出55种化合物有可能是其发挥作用的主要成分，发现逍遥散中未曾报道的7种可能具有神经保护作用的化合物，并验证出苯乙醛具有神经保护作用且此前从未报道。本研究成果将为阐明逍遥散神经保护作用的物质基础提供参考，也为拓展逍遥散的临床应用提供数据支撑。

Abstract:

Objective To study the neuroprotective effect of Xiaoyaosan and its chemical composition，and to identify the active ingredients in Xiaoyaosan responsible for neuroprotection.Methods Xiaoyaosan was extracted using water，and the chemical composition of extract was analyzed using high-resolution mass spectrometry and liquid chromatography. Moreover，we prepared Xiaoyaosan-containing serum and established a corticosterone（CORT，400 μmol/L） -induced PC12 cell injury model to evaluate the impact of the drug-containing serum on the proliferation rate of PC12 cells. Then we used SIMCA software to analyze the correlations between cell proliferation rate and drug components to identify the compounds in Xiaoyaosan that may have neuroprotective effects.Results Compared with the model group，the drug-containing serum could reduce the cell injury induced by CORT and promote cell proliferation（P=0.000）. Analysis of liquid chromatography-mass spectrometry data in software showed that the main medicinal compounds were：paeoniflorin，glycyrrhizic acid，liquiritin，rosmarinic acid，gallic acid，2-phenylacetaldehyde，（15Z）-9，12，13-trihydroxy-15-octadecenoic acid，salvianolic acid B，and licorice saponin G2. The experimental results of individual compounds showed that phenylacetaldehyde was effective in neuroprotection（P=0.000）.Conclusion This research proves that Xiaoyaosan has neuroprotective effect. Fifty-five compounds were identified as potential ingredients involved in neuroprotection. Seven compounds，previously unreported and possessing potential neuroprotective properties，were identified in Xiaoyaosan. In addition，we demonstrate that phenylacetaldehyde has a neuroprotective effect，which has not been reported. The results of this study provide a reference for elucidating the material basis of the neuroprotective effect of Xiaoyaosan，and also provide data support for expanding the clinical application of Xiaoyaosan.