黄芪皂苷Ⅱ通过阻断NOX/ROS/AKT/mTOR信号通路抑制低氧诱导的大鼠肺动脉平滑肌细胞增生  

AstragalosideⅡinhibits the proliferation of rat pulmonary artery smooth muscle cells induced by hypoxia via blocking NOX/ROS/AKT/mTOR signaling pathway

作　　者：金海峰[1,2] 姚立杰 杜凤霞[3] 姜杨[1] 李雪梅[4] 张彧婷 王冠[1] 刘吉成 周忠光 JIN Haifeng;YAO Lijie;DU Fengxia;JIANG Yang;LI Xuemei;ZHANG Yuting;WANG Guan;LIU Jicheng;ZHOU Zhongguang(Department of Anatomy,Qiqihar Medical University,Qiqihar 161006;Qigihar Institute of Medical and Pharmaceutical Sciences,Qiqihar Medical University,Qiqihar 161006;Department of Etiology,Qiqihar Medical University,Qiqihar 161006;Experiment and Practice Training Center,Qiqihar Medical University,Qiqihar 161006;Basic Discipline of Integrated Chinese and Western Medicine,Heilongjiang University of Chinese Medicine,Harbin 150000,China)

机构地区：[1]齐齐哈尔医学院解剖教研室,黑龙江齐齐哈尔161006 [2]齐齐哈尔医学院医药科学研究院,黑龙江齐齐哈尔161006 [3]齐齐哈尔医学院病原教研室,黑龙江齐齐哈尔161006 [4]齐齐哈尔医学院实验实训中心,黑龙江齐齐哈尔161006 [5]黑龙江中医药大学中西医结合基础,黑龙江哈尔滨150000

出　　处：《细胞与分子免疫学杂志》2021年第3期219-224,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：黑龙江省省属高等学校基本科研业务费科研项目(2018-KYYWF-0115);黑龙江省博士后资助项目(LBH-Z17213);齐齐哈尔医学科学院基金(QMSI2020M-04);齐齐哈尔医学院院内科研基金(QY2016M-02)。

摘　　要：目的探讨黄芪皂苷Ⅱ(AS-Ⅱ)对低氧诱导的肺动脉平滑肌细胞(PASMC)过度增生的抑制作用及相关机制。方法将大鼠原代PASMC分为常氧组、低氧组、低氧联合(20、40、80)μmol/L AS-Ⅱ处理组、低氧联合还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶(NOX)抑制剂VAS2870处理组,在常氧(210 mL/L O_(2))或低氧(20 mL/L O_(2))条件下培养24 h。利用CCK-8法检测细胞增殖活性,二氯二氢荧光素二乙酸酯(DCFH-DA)荧光探针检测细胞内活性氧(ROS),Western blot法检测细胞蛋白激酶B(AKT)、磷酸化的AKT(p-AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)、磷酸化的mTOR(p-mTOR)、增殖细胞核抗原(PCNA)以及NOX1、NOX4的蛋白表达。结果与常氧组相比,低氧组PASMC明显增生,细胞内ROS含量显著升高,PCNA、p-AKT、p-mTOR、NOX1、NOX4蛋白均表达增加。AS-Ⅱ处理抑制低氧诱导的PASMC增生,降低细胞ROS水平,并抑制PCNA、p-AKT、p-mTOR、NOX1、NOX4的蛋白表达。VAS2870处理后也有类似改变。结论AS-Ⅱ抑制低氧诱导的PASMC增生,可能与阻断NOX/ROS/AKT/mTOR信号通路相关。Objective To investigate the inhibitory effect of astragalosideⅡ(AS-Ⅱ)on the proliferation of pulmonary artery smooth muscle cells(PASMCs)induced by hypoxia and its relevant mechanism.Methods Rat primary PASMCs were divided into normoxia group,hypoxia group,hypoxia combined with 20,40,80μmol/L AS-Ⅱtreated groups,hypoxia combined with nicotinamide adenine dinucleotide phosphate(NADPH)oxidase(NOX)inhibitor VAS2870 treated group,and then cultured either in normoxic(210 mL/L O_(2))or hypoxic(20 mL/L O_(2))condition for 24 hours.The proliferation of PASMCs was detected by CCK-8 assay.The level of intracellular reactive oxygen species(ROS)was detected by DCFH-DA staining.Protein kinase B(AKT),phospho-AKT(p-AKT),mammalian target of rapamycin(mTOR),phospho-mTOR(p-mTOR),proliferating cell nuclear antigen(PCNA),NOX1 and NOX4 protein expression were assessed by Western blotting.Results In the hypoxia group,the proliferation of PASMCs,level of intracellular ROS,protein expression of PCNA,p-AKT,p-mTOR,NOX1 and NOX4 increased significantly compared with those in the normoxia group.However,AS-Ⅱtreatment inhibited hypoxia-induced PASMCs proliferation,decreased the level of intracellular ROS,and suppressed protein expression of PCNA,p-AKT,p-mTOR,NOX1 and NOX4.Moreover,VAS2870 treatment lead to similar changes.Conclusion AS-Ⅱcan inhibit the proliferation of PASMCs induced by hypoxia,which may be associated with the blocking of NOX/ROS/AKT/mTOR signaling pathway.

关 键 词：黄芪皂苷Ⅱ(AS-Ⅱ) 缺氧 肺动脉平滑肌细胞(PASMC) 活性氧(ROS) NADPH氧化酶(NOX) 

分 类 号：R285.5[医药卫生—中药学] R543.2[医药卫生—中医学]