微粒型灵芝孢子β-葡聚糖重塑免疫抑制微环境增强吉西他滨抗肺癌作用  

Micro-particulate Ganoderma lucidum sporeβ-glucanin enhances the antitumor activity of gemcitabin via remodeling immunosuppressive microenvironment in Lewis lung cancer

作　　者：陈菲菲 李畅 罗毅[1] 魏梦佳丽 马倩 宋捷[1,2] 封亮 贾晓斌 谭晓斌[1,2] CHEN Fei-fei;LI Chang;LUO Yi;WEI Meng-jia-li;MA Qian;SONG Jie;FENG Liang;JIA Xiao-bin;TAN Xiao-bin(Affiliated Hospital of Integrated Traditional Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210028,China;Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica,Jiangsu Provincial Academy of Chinese Medicine,Nanjing 210028,China;School of Traditional Chinese Pharmacy,China Pharmaceutical University,Nanjing 211198,China)

机构地区：[1]南京中医药大学附属中西医结合医院,江苏南京210028 [2]江苏省中医药研究院,国家中医药管理局中药释药系统重点研究室,江苏南京210028 [3]中国药科大学中药学院,江苏南京211198

出　　处：《药学学报》2021年第7期1988-1998,共11页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81773947);江苏省医学创新团队项目(CXTDB2017003);南京中医药大学自然科学基金资助项目(XZR2020027).

摘　　要：吉西他滨(gemcitabine,GEM)为临床治疗非小细胞肺癌的常用药物。然而,用其反复化疗后依然肿瘤迁延,治疗效果不佳,究其原因是化疗使得机体免疫力下降,肿瘤微环境(tumor microenvironment,TME)中介导免疫逃逸的细胞聚集,T细胞耗竭,使得整体偏向于免疫抑制状态,导致了肿瘤的发展。本研究中的动物实验获得江苏省中医药研究院伦理委员会的批准。在体外实验中观察了微粒型灵芝孢子粉β-葡聚糖(micro-particulate Ganoderma lucidum spore β-glucan,PGSG)对巨噬细胞的免疫调节作用,同时建立了小鼠皮下Lewis lung cancer (LLC)模型,口服PGSG联合GEM化疗给药以观察抗LLC肺癌的作用;流式细胞术分析小鼠肿瘤与脾脏中辅助性T细胞-1 (helper T cell-1,Th1)与细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)的比例,以及髓源性抑制细胞(myeloid-derived suppressor cells,MDSC)、肿瘤相关巨噬细胞(tumor-associated macrophages,TAM)与调节性 T细胞(regulatory cells,Tregs)的比例。结果显示,PGSG上调了巨噬细胞CD40、CD86、CD80和主要组织相容性抗原(major histocompatibility antigens complex,MHC-Ⅱ)的表达,增强其对中性红的吞噬能力,增加炎症因子的释放。在体内实验中发现,与单独化疗相比,联合给药能够明显延缓小鼠肿瘤发展,显著降低肿瘤与脾脏中MDSC (CD11b+Gr-1-PE+)、M-MDSC(CD11b+Ly6G-Ly6Chigh)、Tregs (CD4+Foxp3+)和 M2 (F4/80+CD206+)细胞比例,同时 Th1 (CD4+IFN-γ+T细胞)与 CTL(CD8+IFN-γ+T细胞)明显增加。这提示PGSG能够重塑免疫抑制微环境,增强吉西他滨的抗肺癌效果。Gemcitabine(GEM)is a commonly used drug in the clinical treatment of non-small cell lung cancer.Due to the accumulation of cells mediating immune escape and T cell depletion after chemotherapy,tumor microenvironment(TME)tends to be immunosuppressive status,which ultimately leads to tumor metastasis.The experimental protocol was approved by the Medical Laboratory Animal Ethics Committee of Jiangsu Provincial Academy of Chinese Medicine.Therefore,we observed the immunomodulatory effects of micro-particulate Ganoderma lucidum sporeβ-glucan(PGSG)on macrophages in vitro experiments.Next,mice subcutaneous Lewis lung cancer models were established to observe the anti-tumor effects of PGSG through oral administration of PGSG combined with GEM.Flow cytometry analysis was used to analyze the ratio of anti-tumor T cells in tumors and spleen,as well as the proportion of myeloid-derived suppressor cells(MDSC),tumor-associated macrophages(TAM)and regulatory cells(Tregs).The results showed that PGSG can up-regulate the expression of major histocompatibility antigens(MHC-II),CD40,CD86 and CD80 on the surface of macrophages,enhance the ability to phagocytosis of neutral red and further mediate the release of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interleukin-4(IL-4)and interleukin-10(IL-10).In vivo experiments,combined administration can significantly decrease the volume and weight of tumors,reduce the ratio of MDSC(CD11b+Gr-1+),M-MDSC(CD11b+Ly6G-Ly6Chigh)and Treg(CD4+Foxp3+).At the same time,PGSG promoted the conversion of M2(F4/80+CD206+)to M1(F4/80+MHC-II+)and enhanced the response of helper T cell-1(Th1)(CD4+IFN-γ+)and cytotoxic T lymphocyte(CTL)(CD8+IFN-γ+),which is of great significance for killing tumors.These results suggest that PGSG can regulate innate and adaptive antitumor immune responses,reshape the immunosuppressive microenvironment and enhance the anti-lung cancer effect of GEM.

关 键 词：灵芝孢子 Β-葡聚糖 吉西他滨 肿瘤微环境 肿瘤免疫 

分 类 号：R285[医药卫生—中药学]