细胞膜固相色谱法筛选夏枯草抗人乳腺癌细胞效应成分

Screening of Cell Components from Prunella Vulgaris Against Human Breast Cancer Cells by Cell Membrane Solid Phase Chromatography

目的应用细胞膜固相色谱法(CMC)筛选夏枯草中抗人乳腺癌细胞MDA-MB-231的效应成分。方法采用人乳腺癌细胞MDA-MB-231与夏枯草提取液共培养,让人乳腺癌细胞MDA-MB-231的细胞膜上靶点与夏枯草提取液成分结合,洗去未结合的其他成分,再用磷酸盐缓冲液(pH=4.0)使细胞膜靶点失活,让结合于细胞膜上的夏枯草提取液成分解离,解离液经萃取后用液质联用仪进行测定与解析,并用CCK8法检测结合于细胞膜上的成分抗人乳腺癌细胞活性。结果通过细胞膜固相色谱法检测出结合于MDA-MB-231细胞膜上的成分有芦丁、迷迭香酸,CCK8法检测结果显示,芦丁、迷迭香酸均在实验浓度对MDA-MB-231细胞有明显抑制作用(P<0.05,P<0.01)。结论夏枯草中抗人乳腺癌细胞活性的效应成分为芦丁、迷迭香酸,芦丁、迷迭香酸可以作为夏枯草的质量控制成分。

OBJECTIVE To screen the active components in Prunella vulgaris for antibreast cancer cells MDA-MB-231 by rat striatum cell membrane chromatography(CMC) method. METHODS Antibreast cancer cells MDA-MB-231 were cultured with the Prunellae Spica extract, so that cell membrane targets of antibreast cancer cells MDA-MB-231 were combined with the Prunellae Spica extract ingredients.The other ingredients that were not bound were washed away, and then the cell membrane targets were inactivated with the phosphate buffer(pH=4.0), which was bound to the cell membrane so that Prunellae Spica combined components were dissociated from cell membrane.Dissociation fluid after extraction was determinated and analyzed by liquid chromatography-Tandem mass spectrometry and CCK8 assay was used to detect the anti-human breast cancer cell activity on the cell membrane. RESULTS The components bound to the MDA-MB-231 cells were rutin and rosmarinic acid by cell membrane solid phase chromatography.The results of CCK8 method showed that rutin and rosmarinic acid had significant inhibitory effects on the MDA-MB-231 cells at the experimental concentration(P<0.05, P<0.01). CONCLUSION The effective components of Prunella vulgaris against human breast cancer cells are rutin and rosmarinic acid, which can be used as the quality control components of Prunella vulgaris.