复方黄黛片含药鼠血清对HL-60细胞的作用研究  

A Study of the Effect of Rat Serum Containing Compound Realgar Natural Indigo Tablet on HL-60 Cells

作　　者：侯美辰 张英杰[2] 李旭[3] 姜爽 樊磊[2] 史丹 王晓波[2] Hou Meichen;Zhang Yingjie;Li Xu;Jiang Shuang;Fan Lei;Shi Dan;Wang Xiaobo(Pharmaceutical College,Liaoning University of Traditional Chinese Medicine,Dalian 116600.China;The 967th Hospital of Joint Logistics Force,Dalian 116021,China;Dalian Medical University,Dalian 116044,China)

机构地区：[1]辽宁中医药大学药学院,辽宁大连116600 [2]中国人民解放军联勤保障部队第九六七医院,辽宁大连116021 [3]大连医科大学,辽宁大连116044

出　　处：《亚太传统医药》2020年第5期29-33,共5页Asia-Pacific Traditional Medicine

基　　金：辽宁省自然科学基金项目(20180551026)。

摘　　要：目的:临床上复方黄黛片对急性早幼粒细胞白血病具有良好疗效。中药组方未经体内代谢直接用于体外实验不具备说服性。依照血清药理学方法制备复方黄黛片含药血清观察对HL-60细胞的影响,证实复方黄黛片血清用药诱导急性早幼粒细胞凋亡的可信性,提供中药组方作用于体外实验的新途径。方法:制备复方黄黛片混悬液,大鼠灌胃后以腹主动脉取血方式提取含药血清,电感耦合等离子体发射光谱仪测定血清中砷的浓度。不同浓度含药血清,测定对HL-60细胞凋亡的影响。结果:含药血清砷浓度为10μg·L-1。光镜下含药血清增加细胞生长趋势越来越松散,边缘模糊甚至破碎。CCK-8法细胞抑制率、流式细胞仪测总凋亡率都随着含药浓度升高而增大。荧光显微镜观察到含药血清浓度越高细胞存活越少。血清浓度增加凋亡蛋白Bcl-2、Caspase-3和Caspase-8表达下调,Bax、Cleaved Caspase-3和Cleaved Caspase-8表达上调。结论:HL-60细胞对含药血清浓度有一定的依赖作用,复方黄黛片含药血清添加量增多,浓度升高,抑制HL-60细胞增殖作用增强,诱导细胞凋亡程度越显著。Objective:In clinic,compound realgar natural indigo tablet(CRNIT)has a good effect on acute promyelocytic leukemia.It is not persuasive to use Chinese herbal formula directly in vitro without metabolism in vivo.According to the method of serum pharmacology,serum containing CRNIT was prepared to observe the effect on HL-60 cells.To confirm the credibility of serum application of CRNIT in inducing acute promyelocyte apoptosis.In terms of scientific research,it provides a new way for traditional Chinese medicine to act on in vitro experiments.Methods:Preparation of CRNIT suspension.Extraction of serum containing drugs by abdominal aorta after intragastric administration in rats.The concentration of arsenic in serum was determined by ICP-OES.The effect of different concentration of serum containing drugs on the apoptosis of HL-60 cells.Results:The concentration of arsenic in serum containing drugs was 10μg·L-1.Under the light microscope,the cell growth trend of the increase of serum containing drugs is more and more loose,the edge is fuzzy or even broken.The inhibition rate of CCK-8 method and the total apoptosis rate measured by flow cytometry increased with the increase of drug concentration.It was observed by fluorescence microscope that the higher the concentration of serum containing drugs,the less the cells survived.With the increase of serum concentration,the expression of Bcl-2,Caspase-3 and Caspase-8 protein decreased,Bax,Cleaved Caspase-3 and Cleaved Caspase-8 protein increased.Conclusion:HL-60 cells have a certain dependence on the concentration of serum containing drugs.The higher the dose and concentration of serum containing CRNIT,the stronger the inhibition of HL-60 cell proliferation,and the more significant the degree of apoptosis.

关 键 词：复方黄黛片 血清药理学方法 HL-60细胞 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]