黄芪甲苷调控p62-Nrf2通路对抗小鼠淋巴细胞白血病耐药株的机制研究  

Mechanism research of astragaloside Ⅳ on drug-resistant strain of mouse lymphocytic leukemia based on p62-Nrf2 pathway

作　　者：王晓玲 杨小娟 郑倩倩 侯梦雪 王旭 汪涛 WANG Xiaoling;YANG Xiaojuan;ZHENG Qianqian;HOU Mengxue;WANG Xu;WANG Tao(College of Integrated Traditional and Western Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)

机构地区：[1]天津中医药大学中西医结合学院,天津301617

出　　处：《天津中医药》2021年第12期1609-1613,共5页Tianjin Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81503401,81673732);天津自然科学基金项目(18JCYBJC27100);研究生科研创新项目(YJSKC20201038,ZXYCXLX201910202010)。

摘　　要：[目的]基于p62-核因子红细胞样2相关因子2(Nrf2)通路探讨黄芪甲苷(ASTⅣ)对抗小鼠淋巴细胞白血病耐药株的机制。[方法]复制白血病细胞耐药模型,将细胞分为6组,即空白组(blank)、模型组(model)、黄芪甲苷组(ASTⅣ)、盐酸维拉帕米(VER)、Nrf2通路抑制剂(ML385)组及抑制剂与黄芪甲苷组(ML385+ASTⅣ),药物干预48 h。细胞计数试剂盒-8(CCK-8)检测耐药细胞对顺铂的敏感性,流式细胞仪检测细胞内活性氧(ROS)变化,明确ASTⅣ对耐药细胞的影响;流式细胞仪检测细胞周期及凋亡率变化,荧光实时定量聚合酶链式反应(Real time PCR)、蛋白免疫印迹(Western Blot)检测p62-Nrf2通路p62、Nrf2、血红素氧化酶-1(HO-1)、B淋巴细胞瘤-2基因(bcl-2)、bcl-2相关蛋白X(bax)及半胱氨酸蛋白酶-3(Caspase-3)等表达变化明析ASTⅣ对耐药细胞的作用机制。[结果]ASTⅣ通过下调p62-Nrf2通路节点基因p62、HO-1、bcl-2/bax,上调Caspase-3的表达量,降低耐药细胞ROS水平,增加其对顺铂敏感性,使其增殖受抑,细胞周期呈现DNA合成期(S)、合成后期(G2)/有丝分裂期(M)阻滞,凋亡率升高。[结论]ASTⅣ具有逆转白血病耐药的功效,可能与p62-Nrf2通路调控相关,但细胞最终命运受多条通路共同调控仍需探讨。[Objective]To investigate the effect of astragalosideⅣ/ASTⅣon drug-resistant strain of mouse lymphocytic leukemia based on p62-Nrf2 pathway.[Methods]The drug-resistant leukemia model was replicated in vitro,and drug-resistant cells were divided into 6 groups,namely blank group,model group,ASTⅣgroup,VER group,ML385 group,and ML385+ASTⅣgroup.They were subsequently treated with drugs for 48 hours.CCK-8 assay and ROS quantification were performed to observe the effects of ASTⅣon drug-resistance.To analyze the reversal mechanism of ASTⅣon drug resistance,Flow cytometry was performed to detect the changes of cell cycle and apoptosis rate,and real time RT-PCR and Western blot were performed to detect the expression of p62,Nrf2,HO-1,bcl-2,bax and Caspase-3 in p62-Nrf2 pathway.[Results]ASTⅣcould down regulate the expression of p62,HO-1,bcl-2/bax,but up regulate the expression of Caspase-3,resulting in that drug-resistant cells were more sensitive to cisplatin because of the decreased ROS level.The proliferation of drug-resistant cells was inhibited.The cell cycle showed S-phase,G2,M phase arrest and the apoptosis rate increased.[Conclusion]ASTⅣcan have the effect of reversing drug-resistance of leukemia cells,which may be related to the regulation of p62-Nrf2 pathway.However,the co-regulation of multiple pathways in cell fate still needs to be discussed.

关 键 词：p62-Nrf2通路 黄芪甲苷 化疗耐药 

分 类 号：R273[医药卫生—中西医结合]