桦褐孔菌提取物抗乳腺癌活性部位筛选及成分分析

Screening of Anti-Breast Cancer Active Fractions in Inonotus obliquus and Its Component Analysis

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76.17μg/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 76.17μg/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.