摘要
目的探讨东莨菪内酯在大鼠肝微粒体孵育体系中的代谢产物及其含药血清对大鼠肝星状细胞的影响。方法应用高分辨液质联用技术对东莨菪内酯在大鼠肝微粒体孵育体系中的代谢产物进行分析,并观察其含药血清对大鼠肝星状细胞的影响。采用ACQUITY UPLC HSS T3色谱柱(100 mm×2.1 mm内径1.8μm)和0.1%甲酸水溶液-0.1%甲酸乙腈流动相进行梯度洗脱,通过高分辨的二级质谱分析东莨菪内酯的代谢产物。制备各组大鼠含药血清分别运用Annexin V-APC单染法和MTT法检测含药血清诱导大鼠HSC凋亡及抑制其增殖的情况。结果通过大鼠肝微粒体孵育体系统初步鉴定了3个东莨菪内酯的代谢产物,提示其主要代谢途径为脱甲基化和葡萄糖醛酸化反应。含药血清有较弱的诱导大鼠HSC凋亡、抑制HSC增殖作用。结论结合先前的研究,东莨菪内酯的抗肝纤维化作用不直接作用于HSC,应结合其代谢产物情况,进一步进行其作用机制的深入研究,以明确其作用靶点。
Objective To investigate the metabolites of scopoletin using a rat liver microsomal incubation system and the effects of drug-containing serum on rat hepatic stellate cells(HSC).Methods The metabolites of scopoletin from the rat liver microsomal incubation system were analyzed by high-performance liquid chromatography-mass spectrometry(HPLC-MS)and the effects of drug-containing serum on rat HSC were observed.The metabolites of scopoletin were analyzed by high-resolution second-order mass spectrometry with an ACQUITY UPLC HSS T3 column(100 mm×2.1 mm i.d.d.,1.8μm)and 0.1%formic acid aqueous solution-0.1%formic acid acetonitrile mobile phase.Annexin V-APC single staining and MTT method were used to observe the induction of apoptosis and inhibition of proliferation of HSC induced by drug-containing serum.Results Three metabolites of scopolactone were identified with the rat liver microsome incubation system,which indicated that the main metabolic pathway was demethylation and glucuronidation.The drug-containing serum had a weak effect on inducing HSC apoptosis and inhibiting HSC proliferation.Conclusions Combined with previous studies,the anti-fibrotic effects of scopolactone do not directly affect HSC,and further study is needed to clarify the possible mechanism.
作者
杨增艳
张颖
李嘉
陈少锋
黄鑫
靳洪涛
YANG Zengyan;ZHANG Ying;LI Jia;CHEN Shaofeng;HUANG Xin;JIN Hongtao(Guangxi International Zhuang Medicine Hospital,Nanning 530201,China;Guangxi Institute of Chinese Medicine&Pharmaceutical Science,Nanning 530022;Institute of Materia Medica,Chinese Academy of Medical Sciences,Beijing 100050)
出处
《中国比较医学杂志》
CAS
北大核心
2020年第2期9-14,共6页
Chinese Journal of Comparative Medicine
基金
国家自然科学基金课题(81760764,81260683)
中国医学科学院医学与健康科技创新工程项目(2017-I2M-1-011).
作者简介
杨增艳(1975—),男,副主任药师,硕士生导师,研究方向:主要从事中药、民族药研究与开发工作。E-mail:149885961@qq.com;通信作者:靳洪涛(1976—),男,研究员,硕士生导师,研究方向:毒性机制研究与有效干预。E-mail:jinhongtao@imm.ac.cn
