黑参皂苷的提取纯化及其对MGC-803细胞凋亡的影响  

Extraction and Purification of Black Ginseng Saponins and the Effect on Apoptosis of MGC-803 Cells

作　　者：郭晓萌 魏婷[1] 张焱 GUO Xiao-meng;WEI Ting;ZHANG Yan(College of Food Science and Engineering,Tianjin University of Science&Technology,Tianjin 300457,China)

机构地区：[1]天津科技大学食品科学与工程学院,天津300457

出　　处：《食品研究与开发》2023年第4期49-57,共9页Food Research and Development

摘　　要：为了探究黑参皂苷的抗肿瘤作用,将生晒参在高温高湿条件下制备成黑参,采用乙醇超声辅助法提取皂苷,并用大孔树脂纯化,液相质谱联用技术(liquid chromatograph-mass spectrometer,LC-MS)测定人参和黑参的皂苷类成分。采用MTT法、流式细胞术、蛋白质印迹法(Western Blot)探讨黑参皂苷对胃癌MGC-803细胞的抑制作用及诱导其凋亡机制。经大孔树脂纯化后黑参皂苷含量为89.02%。LC-MS分析黑参具有14种单体皂苷,其中Rg3、Rg5为稀有皂苷。在对MGC-803细胞活性方面,人参皂苷24 h的IC50为296.64μg/mL,黑参皂苷为141.25μg/mL,且用浓度为200μg/mL的黑参皂苷处理细胞24 h后,其凋亡率为66.96%,线粒体膜电位下降至23.45%,活性氧(reactive oxygen species,ROS)水平升高至74.65%。Western Blot结果表明黑参皂苷处理组Bax、Cyt-C、Caspase-3、Caspase-9的表达量均增加,Bcl-2的表达量降低。研究表明,制备的黑参产生了稀有皂苷。黑参皂苷有更强的抗肿瘤活性,细胞凋亡机制是通过Bax、Bcl-2蛋白表达的改变导致线粒体膜电位降低,释放Cyt-C,Caspase-3、Caspase-9被激活而裂解使胃癌MGC-803细胞发生凋亡。This study aim to explore the anti-tumor effect of black ginseng saponins(BGS).Specifically,sundried ginseng was used to prepare black ginseng in the environment with high temperature and high humidity.Saponins were extracted by ultrasound-assisted extraction with ethanol and purified with macroporous resin.Liquid chromatography-mass spectrometry(LC-MS)was employed determine the constituents of ginsenosides and BGS.MTT assay,flow cytometry,and Western blotting(WB)were used to investigate the viability of MGC-803 cells and the pro-apoptosis mechanism of BGS.After the purification with macroporous resin,the content of BGS stood at 89.02%.LC-MS suggested that black ginseng had 14 monomer saponins,among which Rg3 and Rg5 were rare saponins.The half-maximal inhibitory concentration(IC50)of ginsenosides and BGS on viability of MGC-803 cells in 24 h was 296.64μg/mL and 141.25μg/mL,respectively.After the treatment of MGC-803 cells with BGS at 200μg/mL for 24 h,the total apoptosis rate of 66.96%,mitochondrial membrane potential of 23.45%,and reactive oxygen species(ROS)level of 74.65%were monitored.The expression of Bax,Cyt-C,Caspase-3,and Caspase-9 increased and the expression of Bcl-2 decreased in the saponin treatment group.The study showed that rare saponins formed in the processing of sun-dried ginseng into black ginseng.BGS had stronger anti-tumor activity,and the pro-apoptosis effect was achieved by regulating the expression of Bax and Bcl-2 proteins to decrease mitochondrial membrane potential,induce the release of Cyt-C,and activate Caspase-3 and Caspase-9.

关 键 词：黑参 人参 皂苷 抗肿瘤 胃癌 凋亡 

分 类 号：R285[医药卫生—中药学]