黄芪多糖调节程序性死亡分子1/程序性死亡配体1信号通路对非霍奇金淋巴瘤发展和免疫逃逸的影响  

Effects of Astragalus Polysaccharide on the Development and Immune Escape of Non-Hodgkin's Lymphoma by Regulating Programmed Death Molecule 1/Programmed Death Ligand 1 Signaling Pathway

作　　者：陈姣敏 许卫星[1] 张薇 CHEN Jiao-Min;XU Wei-Xing;ZHANG Wei(Dept.of Hematology,Cangzhou Central Hospital,Cangzhou 061001 Hebei,China)

机构地区：[1]沧州市中心医院血液内一科,河北沧州061001

出　　处：《广州中医药大学学报》2024年第7期1865-1872,共8页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：河北省中医药管理局科研计划项目(编号:2023454)。

摘　　要：【目的】观察黄芪多糖对非霍奇金淋巴瘤(NHL)的治疗作用及机制。【方法】培养WSU-DLCL2、BJAB、SP53等3种NHL细胞,给予不同浓度(0、5、10、50、100、200、400μmol/L)黄芪多糖处理,应用细胞计数试剂盒8(CCK-8)法检测细胞活力,筛选黄芪多糖最佳浓度、对黄芪多糖敏感的细胞。将WSU-DLCL2细胞分为对照组、黄芪多糖组、BMS-1[程序性死亡分子1(PD-1)/程序性死亡配体1(PD-L1)通路抑制剂]组、黄芪多糖+BMS-1组,检测各组细胞菌落形成、凋亡、迁移和侵袭情况,Western Blot法检测PD-1/PD-L1信号通路蛋白表达。WSU-DLCL2细胞和CD8+T细胞共培养后,检测CD8+T细胞百分比、凋亡率和上清液中干扰素(IFN)-γ、肿瘤坏死因子(TNF)-α水平。【结果】后续实验选择200μmol/L黄芪多糖处理WSUDLCL2细胞。与对照组比较,黄芪多糖组、BMS-1组和黄芪多糖+BMS-1组WSU-DLCL2细胞菌落形成数、迁移细胞数、侵袭细胞数,CD8+T细胞凋亡率,及WSU-DLCL2细胞PD-1、PD-L1蛋白表达水平显著下降(P<0.05),WSU-DLCL2细胞凋亡率、CD8+T细胞比例及共培养体系上清液中IFN-γ、TNF-α水平显著升高(P<0.05);与黄芪多糖组、BMS-1组比较,黄芪多糖+BMS-1组WSU-DLCL2细胞菌落形成数、迁移细胞数、侵袭细胞数,CD8+T细胞凋亡率,及WSU-DLCL2细胞PD-1、PD-L1蛋白表达进一步下降(P<0.05),WSU-DLCL2细胞凋亡率、CD8+T细胞比例及共培养体系上清液中IFN-γ、TNF-α水平进一步升高(P<0.05)。【结论】黄芪多糖可抑制NHL细胞增殖、迁移和侵袭,并促进其凋亡,减少免疫逃逸,其机制可能与抑制PD-1/PD-L1信号通路有关。Objective To observe the therapeutic effect and mechanism of astragalus polysaccharide on nonHodgkin's lymphoma(NHL).Methods Three kinds of NHL cells,WSU-DLCL2,BJAB and SP53,were cultured and treated with different concentrations(0,5,10,50,100,200,400 μmol/L) of astragalus polysaccharide.The cell viability was detected by cell counting kit-8(CCK-8) method to screen the optimal concentration of astragalus polysaccharide and the cells sensitive to astragalus polysaccharide.WSU-DLCL2 cells were divided into control group,astragalus polysaccharide group,BMS-1 [programmed death molecule 1(PD-1)/programmed death ligand 1(PD-L1) pathway inhibitor] group,astragalus polysaccharide+BMS-1 group.The colony formation,apoptosis,migration and invasion of each group were detected,and the protein expression of PD-1/PD-L1 signaling pathway was detected by Western Blot.After co-culture of WSU-DLCL2 cells and CD8+ T cells,the percentage and apoptosis of CD8+ T cells,and the levels of interferon(IFN)-γ and tumor necrosis factor(TNF)-α in the supernatant were detected.Results The follow-up experiments selected 200 μmol/L astragaloside to treat WSU-DLCL2 cells.Compared with the control group,the number of WSU-DLCL2 cell colony formation,the number of migrating WSU-DLCL2 cells,the number of invading WSU-DLCL2 cells,the apoptosis rate of CD8+ T cells,and the protein expression levels of PD-1 and PD-L1 in WSU-DLCL2 cells were significantly decreased in the Astragali polysaccharide group,the BMS-1 group and the Astragali polysaccharide+BMS-1 group(P<0.05),and the apoptosis rate of WSU-DLCL2 cells,the proportion of CD8+ T cells,and the levels of IFN-γ and TNF-α in the supernatant of co-culture system were significantly increased(P<0.05);compared with astragali polysaccharide group and BMS-1 group,the number of WSU-DLCL2 cell colony formation,migrating WSU-DLCL2 cells,invading WSU-DLCL2 cells,CD8+ T-cell apoptosis rate,and protein expressions of PD-1,PD-L1 in WSU-DLCL2 cells were further decreased in astragali polysaccharide+BMS-1 grou

关 键 词：黄芪多糖 非霍奇金淋巴瘤 免疫逃逸 程序性死亡分子1(PD-1) 程序性死亡配体1(PD-L1) BJAB细胞 SP53细胞 WSU-DLCL2细胞 

分 类 号：R285.5[医药卫生—中药学]