黄芪多糖调节黑色素瘤小鼠PD-1/PD-Ls分子表达的研究

目的:研究中药黄芪提取物黄芪多糖在黑色素瘤B16-F10细胞荷瘤小鼠体内的抑瘤效应及对共刺激分子PD-1/PD-Ls通路的调节作用,阐明相关的抗肿瘤免疫调节机制。方法:对荷瘤小鼠进行黄芪多糖干预,绘制肿瘤生长曲线并计算抑瘤率;CCK-8法检测荷瘤小鼠外周T淋巴细胞增殖活性;ELISA法检测荷瘤小鼠外周血IL-2、IFN-γ的分泌水平;Real-time PCR法检测荷瘤小鼠脾脏PD-1及肿瘤组织PD-L1、PD-L2在mRNA水平的表达;Western Blot法检测荷瘤小鼠脾脏PD-1及肿瘤组织PD-L1、PD-L2在蛋白水平的表达。结果:黄芪多糖可显著抑制B16-F10细胞在C57BL/6小鼠腋部皮下移植形成肿瘤的生长(P0.01);促进荷瘤小鼠外周T淋巴细胞的增殖(P0.01);升高荷瘤小鼠外周血IL-2、IFN-γ的分泌水平(P0.01);抑制荷瘤小鼠脾组织PD-1mRNA和蛋白的表达(P0.05,P0.01);抑制小鼠肿瘤组织中PD-L2的mRNA表达和PD-L1、PD-L2的蛋白表达水平(P0.01)。结论:黄芪多糖能抑制荷瘤小鼠皮下黑色素瘤的生长,其机制可能与黄芪多糖调节PD-1、PD-Ls分子的表达及相互作用,进而增强小鼠T淋巴细胞抗肿瘤免疫活性有关。

Regulatory Effect of Astragalus Polysacharin on Expression of PD-1/PD-Ls Molecules in Melanoma Mice

Objective： To investigate the anti-tumor effect and the inhibitory effect of Astragalus Polysacharin（ APS） on PD-1 /PD-Ls pathway in rats bearing melanoma B16-F10 cells,and to explore the anti-tumor immunity-regulating mechanism of APS. Methods： The tumor growth curves and the tumor-growth inhibition rates were drawn and calculated through the intervention and observation on rat models. The proliferation activity of splenic T lymphocytes was measured by CCK-8 assay.The secreting levels of IL-2 and IFN-γ in peripheral sera of melanoma-bearing mice were measured by ELISA assay. PD-1,PD-L1 and PD-L2 protein and mRNA expressions in spleen and tumor tissues of melanoma-bearing mice were detected by Western blotting assay and Real-time PCR assay respectively. Results： APS inhibited the tumor growth in melanoma mice（ P 〈0. 01） and increased the proliferation of splenic T lymphocytes（ P 〈0. 01）,and increased secreting levels of IL-2 and IFN-γ in peripheral blood（ P 〈0. 01）. APS also depressed the expression of PD-1 mRNA and protein in spleen（ P 〈0. 05,P 〈0. 01）,as well as inhibited the expressions of PD-L2 mRNA and PD-L1,PD-L2 proteins in tumor（ P 〈0. 01）.Conclusion： APS can significantly inhibit the proliferation and growth activities of B16-F10 cells in C57 BL /6 mouse subcutaneous transplantation models,and its mechanism may be related to regulating the PD-1 and PD-Ls＇ expression,and promoting the immunocompetence of T lymphocytes in mice.