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、400 μmol/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200 μmol/L 黄芪多糖处理WSU-DLCL2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.
Abstract
Objective To observe the therapeutic effect and mechanism of astragalus polysaccharide on non-Hodgkin'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 group(P＜0.05),and the rate of WSU-DLCL2 cell apoptosis,the proportion of CD8+T-cells,and the levels of IFN-γ,TNF-α in the supernatant of co-culture system were significantly further elevated(P＜0.05).Conclusion Astragalus polysaccharide can inhibit NHL cell proliferation,migration and invasion,promote its apoptosis and reduce immune escape,and its mechanism may be related to the inhibition of PD-1/PD-L1 signaling pathway.
国家科技期刊平台
NSTL
万方数据