具有神经细胞保护作用的中药组分快速筛选方法研究

Development of a rapid screening method for discovering neuroprotective components from traditional Chinese medicine

采用二乙酸荧光素(fluorescein diacetate,FDA)标记经L-谷氨酸损伤的SH-SY5Y细胞,并使用细胞荧光显微成像平台检测细胞活力,建立了一种神经细胞保护药物快速筛选方法。将其应用于筛选葛根芩连汤抗神经细胞损伤活性组分,发现C15,D06,D07,E05这4个组分对L-谷氨酸损伤神经细胞具有较好的保护作用。LC-Q/TOF-MS分析鉴定结果表明,葛根芩连汤活性组分的主要成分是大豆苷、3'-甲氧基大豆苷、芹糖甘草苷、白杨素-6-C-L-阿拉伯糖-8-C-D-葡萄糖苷、芹糖异甘草苷、黄芩苷、千层纸素A-7-O-β-D-葡萄糖醛酸苷及汉黄芩苷,提示这些化合物可能具有神经细胞保护作用。

This study developed a method for rapid screening neuroprotective compounds with FDA （fluorescein diacetate） labeled SH-SYSY cells, which was injuried by Glu （L-glutamie acid）. The cell viability was determined by fluorescence automatic microscopy screening system. Then, neuroprotective components from Gegen Qinlian decoction were screened by the proposed method. The results demonstrated that 4 chemical components, C15, DO6, DO7 and E05 from Gegen Qinlian decoction, showed significantly neuroprotective effects. Furthermore, 8 compounds （i. e. daidzin, 3＇-methoxydaidzin, liquiritin apioside, 6-C-L-α-arabinopyranosyl-8- C-/3-D-glucopyranosyl chrysin, isoliquiritin apioside, baicalin, oroxylin-A-7-O-/3-D-glucuronide and wogonoside） were identified from these active components by LC-Q/TOF-MS technology （ liquid chromatography quadrupole time of flight tandem mass spectrometry）. These 8 compounds may be the potential neuroprotective substances in Gegen Qinlian decoction.