鸦胆子苷B与肠道菌群相互作用及对人肺癌A549细胞的抑制作用  

Interaction Between Bruceoside B and Intestinal Flora and Its Inhibitory Effect on Human Lung Cancer A549 Cells

作　　者：石凌钰 王文敏 冯育林[1,2] 杨世林 万阳[1] 陈道峰 温泉[1] SHI Lingyu;WANG Wenmin;FENG Yulin;YANG Shilin;WAN Yang;CHEN Daofeng;WEN Quan(Jiangxi University of Chinese Medicine,Nanchang 330006,China;State Key Laboratory of Innovation Drug and Efficient Energy-saving Pharmaceutical Equipment,Jiangxi University of Chinese Medicine,Nanchang 330006,China;School of Pharmacy,Fudan University,Shanghai 201203,China)

机构地区：[1]江西中医药大学,南昌330006 [2]江西中医药大学创新药物与高效节能降耗制药设备国家重点实验室,南昌330006 [3]复旦大学药学院,上海201203

出　　处：《中国实验方剂学杂志》2024年第13期160-166,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81860684);江西省教育厅项目(GJJ190689);江西省自然科学基金项目(20212ACB206007)。

摘　　要：目的:分析鸦胆子苷B和肠道菌群相互作用,以及其代谢产物对人肺癌A549细胞的抑制活性,探讨鸦胆子苷B对非小细胞肺癌(NSCLC)治疗的价值。方法:采用肠道菌群体外温孵培养法,将鸦胆子苷B与人肠道菌群体外共孵育,通过超高效液相色谱-四极杆-飞行时间串联质谱法(UPLC-Q-TOF-MS)分析代谢转化产物。采用细胞增殖与活性检测法(CCK-8法)检测原型物鸦胆子苷B及其代谢产物对人肺癌A549细胞增殖的影响并计算半数抑制浓度(IC_(50))。取健康雄性大鼠5只,经灌胃给药鸦胆子苷B药液(2 mg·kg^(-1)),连续7 d,分别采集大鼠给药前和给药后的粪便,基于16S rRNA高通量测序分析鸦胆子苷B给药前后肠道菌群变化。结果:鸦胆子苷B可被人肠道菌群经脱糖基水解代谢为鸦胆子苦醇,原型物和转化产物对A549细胞的IC50分别为1755.50、19.57μmol·L^(-1),代谢产物对A549细胞增殖的抑制活性显著优于鸦胆子苷B。肠道菌群分析结果显示,与给药前比较,给药后大鼠肠道菌群α多样性和β多样性未发生明显变化。物种丰度方面,门水平上,鸦胆子苷B降低了放线菌门、变形菌门的物种相对丰度,增加了厚壁菌门、髌骨菌门、蓝细菌门的物种相对丰度;属水平上,鸦胆子苷B降低了葡萄球菌属、气球菌属、嗜冷菌属的物种相对丰度,增加了罗姆布茨菌属、乳杆菌属、狭义梭菌属1、Norank-f-norank-o-Clostridia-UCG-014、苏黎世杆菌属、异杆菌属、糖杆菌念珠菌属的物种相对丰度。功能预测结果显示,与给药前比较,给药后大鼠肠道功能均未发生显著变化。结论:鸦胆子苷B可在肠道菌群作用下脱糖基,转化为苷元鸦胆子苦醇,抗肿瘤活性显著提高。服用鸦胆子苷B不会使肠道菌群结构和功能发生显著变化,造成肠道微生态平衡失调,且给药鸦胆子苷B后出现对NSCLC患者有益的肠道菌群变化。Objective:To explore the interaction between bruceoside B and gut microbiota and the inhibitory activity of its metabolites on human lung cancer A549 cells,and to explore the value of bruceoside B in the treatment of non-small cell lung cancer(NSCLC).Method:Bruceoside B was co-incubated with the human gut microbiota under anoxic conditions in vitro,and ultra high performance liquid chromatographyquadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS)was used to analyze the metabolic transformation products.Cell counting kit-8(CCK-8)assay was performed to determine the effects of bruceoside B and its metabolites on the proliferation of human lung cancer A549 cells and the half inhibitory concentration(IC_(50))was calculated.Five healthy male rats were gavaged with bruceoside B(2 mg·kg^(-1))for 7 days after adaptive feeding.The feces of rats were collected before and after administration.16S rRNA sequencing was used to assess gut microbiota.Result:Bruceoside B was mainly metabolized to brusatol by human gut microbiota,the IC50 of bruceoside B and the conversion product to A549 cells were 1755.50,19.57μmol·L^(-1),respectively,and the conversion product had a better activity at inhibiting A549 cells proliferation than bruceoside B.Additionally,The results of intestinal flora analysis showed no significant differences inαdiversity andβdiversity of gut microbiota after administration.In terms of species abundance,at the phylum level,bruceoside B decreased the relative abundance of Actinobacteriota and Proteobacteria,increased the relative abundance of Firmicutes,Patescibacteria and Cyanobacteria.At the genus level,bruceoside B decreased the relative abundance of Staphylococcus,Aerococcus and Psychrobacter,increased the relative abundance of Romboutsia,Lactobacillus,Clostridium sensu stricto 1,Norank-f-norank-o-Clostridia-UCG-014,Turicibacter,Allobaculum and Candidatus Saccharimonas.The results of functional prediction showed that the gut microbiota functional compositions were relatively stable.Conclusion:Bruce

关 键 词：鸦胆子 鸦胆子苷B 鸦胆子苦醇 肠道菌群 代谢转化 人肺癌A549细胞 非小细胞肺癌(NSCLC) 16S rRNA高通量测序 

分 类 号：R22[医药卫生—中医基础理论] R256.1[医药卫生—中医学] R28R969.1