桦褐孔菌提取物抗乳腺癌活性部位筛选及成分分析

以人乳腺癌MDA-MB-231细胞筛选桦褐孔菌抗乳腺癌活性部位，并对其活性部位的成分进行初步分析。采用MTT法检测发酵液不同萃取部位抗乳腺癌活性；通过细胞形态观察进一步了解药物对细胞的影响；划痕试验检测桦褐孔菌活性部位对细胞迁移的影响；采用LC-MS/MS对活性部位的成分进行分析。结果表明，桦褐孔菌石油醚萃取部位对MDA-MB-231细胞增殖的抑制作用最强，且呈量效和时效关系，作用24 h的IC50值为76.17 μg/mL；细胞形态观察显示石油醚萃取部位对乳腺癌MDA-MB-231细胞的凋亡作用随着药物质量浓度的升高而加强；划痕实验显示，药物对细胞迁移有较强的抑制作用；LC-MS/MS数据初步表征了其中的14个化学成分，初步鉴定了其中的10种成分，这10种成分首次从桦褐孔菌中得到鉴定，虽然其他4个物质未能鉴定，但亦对其相对分子质量和分子式进行了初步表征。证实了石油醚部位是桦褐孔菌抗乳腺癌MDA-MB-231细胞的主要活性部位，能抑制乳腺癌细胞增殖并诱导细胞凋亡，为进一步开发利用桦褐孔菌食药用资源提供科学依据。
To achieve this study，MTT assay is used to measure the anti-breast cancer activity in different extracts from Inonotus obliquus；Change of cellular morphology is observed as the apoptosis detection；Cell scratch test is used to determine the effect of drug on cell migration ；LC-MS/MS is used to analyze the active constituents in drug. MTT shows that the petroleum ether extract fraction has the strongest cell growth inhibitory effect on MDA-MB-231 cells in a dose and time dependent manner（P＜0.05）；the IC50 value for 24 hours is 76.17 μg/mL.Cell morphology observation shows that apoptosis effect of petroleum ether extract fraction is increased in MDA-MB-231 cells in a dose manner. Scratch test shows that the petroleum ether extract fraction has a significant inhibitory effect on cell migration. LC-MS/MS data preliminarily characterizes 14 chemical components，of which 10 components are firstly identified in Inonotus obliquus. Although the other 4 substances have not been identified，the molecular weight and molecular formula have been characterized. The petroleum ether extract fraction is considered as the main anti-breast cancer active fraction of Inonotus obliquus. It can inhibit the growth of MDA-MB-231 cells and induce apoptosis，which provided the scientific basis for further development and utilization of the food and medicinal resources of Inonotus obliquus