摘要
目的:研究维药布亚中金雀花碱对人肝癌细胞HepG-2的抑制增殖及诱导凋亡作用,对金雀花碱3位N进行结构修饰,并对其抗肿瘤活性进行初步测定。方法:MTT法考察金雀花碱对HepG-2细胞的生长抑制作用,倒置显微镜和透射电子显微镜观察细胞形态及细胞超微结构变化,流式细胞仪检测细胞凋亡率。采用亲核取代的方法对活泼氢进行取代,得到金雀花碱衍生物,MTT法比较金雀花碱及其衍生物对HepG-2细胞的增殖抑制作用。结果:金雀花碱对HepG-2细胞的IC_(50)为5.36 mmol/L,可观察到形态变化和凋亡细胞,并出现明显的凋亡峰。~1H-NMR证实化学修饰产物为N-乙酰基金雀花碱,其IC50为7.51 mmol/L。结论:金雀花碱体外具有抑制HepG-2细胞生长的作用,能够诱导HepG-2细胞凋亡。N-乙酰基金雀花碱能够抑制HepG-2细胞的生长,但其作用较金雀花碱弱,表明3位N上修饰链状结构Ac_2O并不能增强金雀花碱的抗肿瘤活性。
Objective： To investigate the inhibition effect and apoptosis inducing effect of cytisine in Sophora alopecuroides L. on human hepatoma cell line HepG-2; to modify structure of cytisine on 3-N; and to detect its anti-tumor activity preliminarily. Methods： MTT method was used to examine the growth inhibition of cytisine on HepG-2 cells. The qualitative researches on apoptosis morphology of HepG-2 cells induced by cytisine are studied by the inverted microscope and transmission electron microscopy. FCM was applied to investigate the apoptosis rate of HepG-2 cells. Nucleophilic substitution method was employed to substitute reactive hydrogen with a cytisine-derivative being produced.MTT test was employed to compare the activity of cytisine and cytisine-derivative. Results： The IC50 level of HepG-2 cells was 5. 36 mmol/L. Inverted microscope and transmission electron microscopy exhibited typical morphologic changes of apoptosis. A typical subdiploid peak was observed by flow cytometry. In the chemical modification experiment,-1H NMR showed that this derivative was N-acetylcytisine,whose IC50 level of HepG-2 cells is 7. 51 mmol/L. Conclusion： The cytisine may inhibit growth and proliferation of HepG2 cell through inducing apoptosis. Though N-acetylcytisine can also significantly inhibit the growth of HepG-2 cells,its inhibition impact was weaker than that of cytisine,which shows that Ac2 O structure in cytisine cannot enhance anti-tumor activity of cytisine.
作者
于蕾
尚东雨
杨经辉
陈志峰
王鑫
姜波
孙永雪
张连芳
季宇彬
YU Lei;SHANG Dongyu;YANG Jinghui;CHEN Zhifeng;WANG Xin;JIANG Bo;SUN Yongxue;ZHANG Lianfang;JI Yubin(Center of Research and Development on Life Sciences and Environmental Sciences,Harbin University of Commerce,Harbin 150076,China;Engineering Reseach Center of Natural Anticaucer Drugs,Ministry of Education,Harbin 150076,China)
出处
《中医药学报》
CAS
2018年第3期43-49,共7页
Acta Chinese Medicine and Pharmacology
基金
黑龙江省普通高等学校肿瘤预防与抗肿瘤药物研究重点实验室开放课题(CPAT-2012003)
哈尔滨市科技局创新人才项目(2014RFQXJ154)
哈尔滨商业大学博士科研启动项目(12DL008)
哈尔滨商业大学科研团队计划(2016TD002)
哈尔滨商业大学青年创新人才支持项目(2016QN057)
