黄芪多糖对H22荷瘤小鼠PD-1抑制剂抗癌能力的影响

Effects of Astragalus Polysaccharide on Inhibitory Ability of PD-1 Inhibitor in H22 Tumor-Bearing Mice

目的:以H22荷瘤小鼠为模型,观察黄芪多糖(APS)对PD-1抑制剂抗癌能力的增强效果并探讨其机制。方法:建立H22荷瘤小鼠模型,分别给予PD-1抑制剂和/或不同浓度APS处理,记录肿瘤生长情况,ELISA法检测肿瘤坏死因子(TNF-α)、白介素1β(IL-1β)、白介素2(IL-2)、干扰素γ(IFN-γ)、白介素4(IL-4)、白介素10(IL-10)、转化生长因子(TGF-β)等血浆细胞因子浓度,Western Blot法检测肿瘤及脾脏组织PD-1表达,流式细胞术检测肿瘤浸润细胞类型。结果:(1)APS及PD-1抑制剂均能抑制H22荷瘤小鼠的肿瘤生长,联合用药效果更为显著,且抑制强度随APS浓度上升而增加。(2)APS联合PD-1抑制剂作用后,血浆TNF-α、IL-1β、IFN-γ、IL-10、TGF-β浓度上升,IL-2、IL-4血浆浓度下降,肿瘤及脾脏组织PD-1蛋白表达下降,肿瘤组织CD_(4)^(+)CD-8细胞和CD_(4)^(+)CD_(8)^(+)细胞浸润比例增加。结论:APS能增强PD-1抑制剂抗H22肝癌能力,可能与调节细胞因子分泌、抑制PD-1表达、增加肿瘤组织淋巴细胞浸润相关。

Objective:To observe the effects of APS on anti-cancer ability of PD-1 inhibitors on H22 tumor-bearing mice model and explore the possible mechanism.Methods:Hepatocellular carcinoma H22 cells were inoculated subcutaneously into BALB/c mice.H22 tumor-bearing mice were randomly divided into different group which treated with PD-1 inhibitor and/or different concentrations of APS.Tumor size was recorded,serum cytokines levels such as TNF-α,IL-1β,IL-2,IFN-γ,IL-4,IL-10,TGF-βwere determined by ELISA,PD-1 expression in tumor and spleen tissue was determined by Western Blot,tumor-infiltrating cell types were determined by flow cytometry.Results:1)Both APS and PD-1 inhibitors could inhibit tumor growth in the H22 tumor-bearing mice,and the effect of combination therapy was more significance and the inhibitory intensity increased with the increasing of APS concentration.2)The serum concentrations of TNF-α,IL-1β,IFNγ,IL-10 and TGF-βwere increased after drug intervention,the serum concentrations of IL-2 and IL-4 were decreased,the expression of PD-1 protein in tumor and spleen tissue was decreased,and the infiltration ratio of CD_(4)^(+)CD-8cells and CD_(4)^(+)CD_(8)^(+) cells in tumor were increased.Conclusion:APS can enhance the inhibitory ability of PD-1 inhibitor on H22 hepatocellular carcinoma,which may be related to regulating cytokines secretion,inhibiting PD-1 expression,and increasing lymphocyte infiltration in tumor tissue.