假马齿苋皂苷诱导K562细胞向巨核细胞分化的作用及相关基因调控机制  

Bacopasides Induces Megakaryocytic Differentiation in K562 Cells and Its Gene Regulation Mechanism

作　　者：蒋心瑞 吴安国 王龙 杨靖 秦大莲 叶云 吴建明 Jiang Xinrui;Wu Anguo;Wang Long;Yang Jing;Qin Dalian;Ye Yun;Wu Jianming(School of Pharmacy,Southwest Medical University,Luzhou 646000;Department of Pharmacy,Affiliated Hospital of Southwest Medical University,Luzhou 646000)

机构地区：[1]西南医科大学药学院,泸州646000 [2]西南医科大学附属医院药学部,泸州646000

出　　处：《中药药理与临床》2019年第5期46-51,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金项目(81774013、81804221);四川省科技厅面上项目(2018JY0237、2019YJ0484、2019JDPT0010)

摘　　要：目的:探索假马齿苋皂苷诱导K562细胞向巨核细胞分化的作用。方法:采用3种假马齿苋皂苷干预K562细胞,应用CCK-8法检测细胞增殖;显微镜观察细胞形态;Giemsa染色观察细胞核倍性;流式细胞术检测巨核细胞表面抗原CD41和CD42b的阳性表达率;高内涵细胞成像分析系统验证细胞的倍性;qRT-PCR技术检测相关基因GATA-1、RUNX-1、NF-E2 mRNA的表达水平。结果:与对照组比较,假马齿苋皂苷Ⅱ与假马齿苋皂苷C诱导K562细胞体积增大;多倍体细胞比例增加;表面抗原CD41和CD42b阳性表达率显著上调,以假马齿苋皂苷Ⅱ的作用最为显著。基因检测结果显示,假马齿苋皂苷Ⅱ干预K562细胞后,胞内GATA-1、RUNX-1与NF-E2 mRNA表达水平显著上调。结论:假马齿苋皂苷Ⅱ具有促进K562细胞向巨核细胞分化作用,其作用机制与上调GATA-1、RUNX-1和NF-E2 mRNA的表达水平有关。Objective:The aim of this study is to investigate the effects of bacopasides on megakaryocyte differentiation of K562 cells.Methods:K562 cells were treated with 3 different concentrations of bacopasideⅡ,bacopasideⅦand bacopasaponin C,then the cell viability was detected by the CCK-8,the morphological change was observed,the ploidy of K562 cells was evaluated by Giemsa staining,the positive expression rates of CD41 and CD42 b were measured by flow cytometry,the K562 cells ploidy was verified by high-content cell imaging analysis system,qRT-PCR was employed to detect the mRNA expression levels of GATA-1,RUNX-1 and NF-E2 in K562 cells.Results:Compared with the control group,after K562 cells were treated with bacopasideⅡand bacopasaponin C,the cellular volumes,the polyploid cell rates and the positive expressions of CD41 and CD42 b on cell membrane were increased.Bacopaside II had the most significant effect.Further ploidy and gene expression assays showed that,bacopasideⅡsignificantly up-regulated the mRNA expression levels of GATA-1,RUNX-1 and NF-E2.Conclusion:BacopasideⅡcan promote K562 cells into megakaryocytic differentiation,and its mechanism may related to up-regulating the mRNA expressions of GATA-1,RUNX-1 and NF-E2.

关 键 词：假马齿苋皂苷 K562细胞 巨核细胞 分化 GATA-1 RUNX-1 NF-E2 

分 类 号：R28[医药卫生—中药学]