基于TLR4信号通路黄芪多糖促树突状细胞抗肿瘤作用的研究  

Anti-tumor effect study of Astragalus polysaccharide promotes dendritic cell through TLR4 signal pathway

作　　者：贾贝田 王丽[2] 崔换天 金昱彤 曹敏 刘海朝 边育红[1] JIA Beitian;WANG Li;CUI Huantian;JIN Yutong;CAO Min;LIU Haizhao;BIAN Yuhong(Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Tianjin Second People’s Hospital,Tianjin 300192,China;Shandong University,Qingdao 266237,China)

机构地区：[1]天津中医药大学,天津301617 [2]天津市第二人民医院,天津300192 [3]山东大学,青岛266237

出　　处：《天津中医药大学学报》2022年第2期218-224,共7页Journal of Tianjin University of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81573707);天津中医药大学中西医结合学院研究生创新基金项目(ZXYCXLX201814)。

摘　　要：[目的]研究基于Toll样受体4(TLR4)信号通路黄芪多糖(APS)活化树突状细胞(DCs)发挥抗肿瘤作用的机制。[方法]将树突状细胞DC2.4随机分为空白(Control)组、APS组、肿瘤坏死因子-α(TNF-α)组、APS+TNF-α组、TAK-242+TNF-α组、TAK-242+APS+TNF-α组、ST2825+TNF-α组及ST2825+APS+TNF-α组,干预72 h后,流式细胞仪检测DCs表型CD80、CD86的表达情况,ELISA法检测DCs对细胞因子白细胞介素(IL)-12分泌水平的影响。将上述干预处理后的DCs和T细胞共培养,ELISA法检测DCs对T细胞活化影响;将活化的细胞毒性T淋巴细胞(CTL)与肿瘤细胞CT 26共培养,全视野细胞成像分析仪检测CTL对肿瘤的杀伤作用。[结果]与Control组相比,APS可显著促进DCs表面分子CD80和CD86的表达以及IL-12的分泌(P<0.05),且APS和TNF-α协同干预时,效果最优。此外,两者协同干预的DCs可显著促进T细胞增殖并分泌IL-2,增强CTL对肿瘤的杀伤(P<0.05)。然而,加入TLR4通路阻断剂TAK-242或ST2825后,APS+TNF-α组DCs表面分子CD80和CD86的表达、IL-12的分泌水平与TNF-α组相比均无显著差异,且两者协同干预的DCs对T细胞增殖、分泌IL-2及肿瘤的杀伤水平亦无显著差异。[结论]APS可能是通过TLR4介导的髓样分化因子(MyD88)通路促进DCs成熟、活化,激活T细胞,进而发挥抗肿瘤的作用。[Objective]The aim of this study is to investigate the anti-tumor effect of Astragalus polysaccharides(APS)activated dendritic cell(DCs)via the TLR4 signaling pathway.[Methods]The DC2.4 were randomly divided into Control group,APS group,TNF-αgroup,APS+TNF-α group,TAK-242+TNF-αgroup,TAK-242+APS+TNF-α group,ST2825+TNF-αgroup and ST2825+APS+TNF-α group.After 72h of intervention,the ratio of CD80+CD86+DCs were detected by flow cytometry,and the effect of DCs on the secretion of cytokine IL-12 were detected by ELISA.Then the DCs in previous step were co-cultured with T cells,and effects of DCs on T cell activation were detected by ELISA;the activated CTL cells were co-cultured with tumor cells CT 26,tumor-cell-killing efficiency was calculated using the full-field cell imaging analyzer.[Results]Compared with the control group,APS can significantly promote the expression of CD80+CD86+DCs and the secretion of IL-12(P<0.05),and the best effects were obtained when APS and TNF-α were synergistically intervened.In addition,the DCs intervened by APS+TNF-α could significantly promote proliferation of T cells,the secretion of IL-2 by T cells and enhance the killing of tumors by CTL(P<0.05).However,after adding the TLR4 pathway blocker TAK-242 or ST2825,the expression of CD80+CD86+DCs and the secretion level of IL-12 in APS+TNF-α group were not significantly different from those in the TNF-α group.Compared with the TNF-α group,there was no significant difference in proliferation of T cells,the level of IL-2 secretion and tumor killing by T cells in APS+TNF-α group with blocker TAK-242 or ST2825.[Conclusion]APS could induce DCs maturation and increase the activating ability of DC on T cells.These effects and mechanism may through TLR4/MyD88 pathway,and APS/TNF-a combination treatment to DCs is the best.

关 键 词：黄芪多糖 树突状细胞 TLR4通路 抗肿瘤 

分 类 号：R285.5[医药卫生—中药学]