青蒿琥酯抑制肾母细胞瘤细胞系SK-NEP-1增殖和促进凋亡与自噬  

Artesunate inhibits proliferation and promotes apoptosis and autophagy of nephroblastoma cell line SK-NEP-1

作　　者：魏建新[1] 房彦乐 鲁雨博[3] 高宇光 郎兴 李静涛[1] 马新生[1] WEI Jianxin;FANG Yanle;LU Yubo;GAO Yuguang;LANG Xing;LI Jingtao;MA Xinsheng(the Second Department of Pediatric Surgery,Handan Central Hospital,Handan 056000;the Third Department of General Surgery,the Second Hospital of Hebei Medical University,Shijiazhuang 050000;Department of Pharmacy,Handan Central Hospital,Handan 056000,China)

机构地区：[1]邯郸市中心医院小儿外二科,河北邯郸056000 [2]河北医科大学第二医院普外三科,河北石家庄050000 [3]邯郸市中心医院药学部,河北邯郸056000

出　　处：《基础医学与临床》2025年第4期493-498,共6页Basic and Clinical Medicine

基　　金：河北省2024年度中医药类科学研究课题计划项目(2024195)。

摘　　要：目的探讨青蒿琥酯(Art)对肾母细胞瘤细胞系(SK-NEP-1)增殖、凋亡和自噬的影响。方法以不同浓度的Art(0、10、20、40和80μmol/L)干预SK-NEP-1细胞,MTT法检测细胞增殖抑制率,筛选最佳干预浓度;将SK-NEP-1细胞分为对照组、Art组、3-MA组(Art+自噬抑制剂,3-甲基腺嘌呤)、Rapa组(Art+自噬激活剂雷帕霉素),EdU、流式细胞测量术分别检测细胞增殖、凋亡;丹磺酰尸胺(MDC)染色法检测细胞自噬;DCFH-DA荧光探针检测细胞活性氧(ROS)的水平;Western blot检测细胞中增殖细胞核抗原(PCNA)、细胞抗凋亡因子B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、微管连接蛋白1轻链3Ⅱ/3Ⅰ(LC3Ⅱ/LC3Ⅰ)、选择性自噬接头蛋白1(p62)和Beclin-1蛋白表达。结果与0μmol/L Art相比,10、20、40和80μmol/L Art处理后SK-NEP-1细胞增殖抑制率逐渐升高(P<0.05),IC 50值为46.881μmol/L,故选取40μmol/L Art进行后续实验。与对照组相比,Art组SK-NEP-1细胞凋亡率、相对自噬荧光强度、ROS水平、Bax、LC3Ⅱ/LC3Ⅰ、Beclin-1、PINK1、Parkin蛋白表达水平显著升高,EdU阳性细胞率、PCNA、Bcl-2、p62蛋白表达水平显著降低(P<0.05);自噬抑制剂3-MA可减弱Art对肾母细胞瘤细胞凋亡和自噬的促进作用,对增殖的抑制作用(P<0.05)。结论Art抑制肾母细胞瘤细胞系的增殖,促进细胞自噬和凋亡。Objective To investigate the effects of artesunate(Art)on the proliferation,apoptosis,and autophagy of nephroblastoma cell line(SK-NEP-1).Methods SK-NEP-1 cells were intervened with different concentrations of Art(0,10,20,40 and 80μmol/L),and MTT method was applied to calculate the cell proliferation inhibition rate to screen the optimal intervention concentration;SK-NEP-1 cells were separated into control group,Art group,3-MA group(Art+autophagy inhibitor,3-methyladenine),and Rapa group(Art+autophagy activator rapamycin).EdU and flow cytometry were applied to detect cell proliferation and apoptosis,respectively;MDC staining was applied to detect autophagy in cells;the level of reactive oxygen species(ROS)in cells was detected by DCFH-DA fluorescent probe;the expression of proliferating cell nuclear antigen(PCNA),anti apoptotic factor B cell lymphomatoma-2(Bcl-2),Bcl-2 associated X protein(Bax),microtubule junction protein 1 light chain 3Ⅱ/3Ⅰ(LC3Ⅱ/LC3Ⅰ),selective autophagy junction protein 1(p62),and benzyl chloride 1(Beclin-1)proteins in cells were detected by Western blot.Results Compared with 0μmol/L Art,the proliferation inhibition rate of SK-NEP-1 cells was gradually increased after 10,20,40 and 80μmol/L Art treatment(P<0.05),and the IC 50 value was 46.881μmol/L,so 40μmol/L Art was selected for follow-up experiments.Compared with the control group,the apoptosis rate,relative autophagy fluorescence intensity,ROS level,Bax,LC3Ⅱ/LC3Ⅰ,Beclin-1,PINK1,and Parkin protein expression levels of SK-NEP-1 cells in the Art group were obviously increased,the EdU positive cell rate,PCNA,Bcl-2,and P62 protein expression levels were obviously reduced(P<0.05);The autophagy inhibitor 3-MA inhibited the promoting effect of Art on apoptosis and autophagy of nephroblastoma cells and inhibit proliferation(P<0.05).Conclusions Art inhibits the proliferation of nephroblastoma cell line SK-NEP-1,and promotes autophagy and apoptosis.

关 键 词：青蒿琥酯 肾母细胞瘤 增殖 凋亡 自噬 

分 类 号：R737.11[医药卫生—肿瘤]