摘要
目的:探究参芪扶正注射液(Shenqifuzheng injection,SFI)联合PD-L1抗体对肿瘤免疫微环境的影响及其疗效研究.方法:构建B16F10-LUC黑色素瘤皮下移植瘤模型,采用免疫组化技术标记抗体 Ki67、CD31、CD8、CD16、CD163、FOXP3、LY6C、LY6G检测肿瘤组织中CD8+T细胞、Treg细胞、NK细胞、MDSCs细胞、中心粒细胞的表达情况,进一步应用流式细胞技术检测脾脏组织中CD11c+、IA/IE+、CD80+细胞的比值,以及肿瘤组织中CD8+T、CD4+T、Treg的比值.结果:免疫组化结果显示,相较于对照组,给药组可以显著抑制肿瘤血管生成、肿瘤细胞增殖,降低免疫抑制细胞因子CD4+T细胞、Treg细胞、MDSCs细胞、中性粒细胞的表达水平,促进CD8、NK的浸润(P＜0.05,P＜0.01).流式结果显示,给药组明显提升了肿瘤组织CD8+T细胞,脾脏中DC细胞的表达水平,抑制CD4+T、Treg细胞的浸润(P＜0.05,P＜0.01).肿瘤体积大小结果显示,给药组明显抑制肿瘤的生长,单用PD-L1抗体抑瘤率优于单用SFI组,联合用药优于单用PD-L1抗体组(P＜0.01).结论:参芪扶正注射液联合PD-L1抗体可以发挥协同抗肿瘤的作用,且可能与增强DC细胞浸润,促进T细胞活化有关,免疫组化结果提示对肿瘤免疫微环境也具有很好的改善.
Abstract
AIM:To investigate of the effect of Shenqifuzheng injection(SFI)combined with PD-L1 antibody on tumor immune microenvironment and its efficacy.METHODS:A subcutaneous transplanta-tion tumor model for B16F10-LUC melanoma was created.The expression of Ki67,CD31,CD8,CD16,CD163,FOXP3,LY6C,LY6G with labeling antibodies was used to detect CD8+T cells,Treg cells,NK cells,MDSCs cells,centrocytes,and granulocytes in the tumor tissues via immunohistochemistry.Flow cy-tometry was used to measure the ratios of CD11c+,IA/IE+,and CD80+cells in splenic tissue,as well as the ratios of CD8+T,CD4+T,and Treg cells in tumor tissue.Additionally,granulocyte count and NK cell expression were analyzed.RESULTS:The immuno-histochemistry results indicate that the drug admin-istration group effectively suppressed tumor angio-genesis and cell proliferation,while decreasing the expression level of immunosuppressive cytokines CD4+T cells,Treg cells,MDSCs and centroblasts.Ad-ditionally,CD8 and NK cell infiltration was promot-ed compared to the control group.The results of the flow analysis demonstrated a significant in-crease in the expression level of CD8+T cells within tumor tissues,as well as inhibition of CD4+T,Treg,and DC cell infiltration within the spleen in the drug administration group.Additionally,the tumor volume analysis indicated that the drug administra-tion group effectively inhibited tumor growth.The flow results illustrate that the group administering treatment exhibited significant increases in CD8+T cell expression levels in tumor tissue and DC cells in the spleen.Furthermore,the treatment effec-tively inhibited the infiltration of CD4+T and Treg cells.The results also indicate that the treatment significantly reduced tumor growth,with the tumor inhibition rate being better with PD-L1 antibody alone than with the SFI group.Additionally,combin-ing drugs resulted in superior results compared to the PD-L1 antibody group alone.CONCLUSION:SFI combined with a PD-L1 antibody can have synergis-tic anti-tumor effects,potentially enhancing DC cell infiltration and promoting T cell activation.Immu-nohistochemistry results indicate a positive impact on the tumor immune microenvironment.
基金项目
国家自然科学基金(81973658)
蚌埠医学院重大科技项目孵化计划(2020byfy001)
安徽省高等学校科研项目(2023AH051972)
蚌埠医科大学自然科学类项目(2023byfy004)
江苏省心脑血管与癌症防控工程研究中心项目(2022)()
江苏省高等学校优秀科技创新团队项目(2023)()
江苏省卫生健康委面上项目(M2022109)
江苏省中医药局项目(YB2020099)
国家科技期刊平台
NSTL
万方数据