丁酸盐在晚期非小细胞肺癌PD-1免疫治疗中发挥协同增效作用  

Butyrate Exerts Synergistic Effects in PD-1 Immunotherapy for Advanced non-small Cell Lung Cancer

作　　者：毛雨珂 任胜楠 冯龄鑫[1] 于壮[1] 李锐[2] MAO Yu-ke;REN Sheng-nan;FENG Ling-xin;YU Zhuang;LI Rui(Department of Oncology,The Affiliated Hospital of Qingdao University,Qingdao University,Qingdao,Shandong,266000,China;Health Management Center,The Affiliated Hospital of Qingdao University,Qingdao University,Qingdao,Shandong,266000,China)

机构地区：[1]青岛大学附属医院肿瘤科,山东青岛266000 [2]青岛大学附属医院健康管理中心,山东青岛266000

出　　处：《现代生物医学进展》2025年第6期961-974,共14页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(82373170);山东省自然科学基金项目(ZR2021MH193);吴阶平医学基金项目(320.6750.2021-02-92)。

摘　　要：目的:探讨晚期非小细胞肺癌(Non-Small Cell Lung Cancer,NSCLC)患者粪便代谢产物丁酸钠在PD-1免疫治疗中发挥的作用及其对肿瘤微环境(Tumor Microenvironment,TME)的相关作用。方法:选择2021年11月至2023年8月于青岛大学附属医院就诊的晚期非小细胞肺癌患者30例,通过气相色谱-质谱联用技术检测不同疗效患者粪便内代谢产物的表达水平。提取小鼠CD8^(+)T细胞,并与小鼠肺癌细胞(LLC)共培养后,使用丁酸钠、PD-1抑制剂单药、PD-1抑制剂联合丁酸盐共处理后。通过细胞活死染色、CCK8实验、Transwell侵袭实验、伤口愈合实验分别检测细胞增殖、凋亡、侵袭和迁移能力的差异。通过ROS流式细胞术、巨噬细胞转型实验、小管形成实验检测细胞产H_(2)O_(2)能力、对TME中巨噬细胞及血管形成的影响。通过体内实验和流式细胞分析术,检测丁酸盐在小鼠转移瘤中对肿瘤免疫微环境的影响。结果:丁酸盐在PD-1抑制剂治疗有效组患者的粪便中的含量显著升高(P<0.05)。使用丁酸钠联合PD-1抑制剂共处理后,CD8^(+)T细胞细胞毒性显著升高,LLC细胞增殖、侵袭和迁移能力显著下降,凋亡能力显著上升(P<0.05)。LLC产H_(2)O_(2)能力显著上升(P<0.05),M1型巨噬细胞比例显著上升(P<0.05),小管形成数量显著下降(P<0.05)。体内小鼠转移瘤CD8^(+)T细胞比例显著上升(P<0.05)。结论:丁酸盐在晚期非小细胞肺癌PD-1抑制剂治疗有效组患者体内高表达,通过体内、体外实验进一步证实了丁酸盐对PD-1抑制剂起到协同增效作用。为克服晚期NSCLC免疫抑制剂治疗耐药提供了新思路。Objective:To investigate the role of sodium butyrate,a fecal metabolite in PD-1immunotherapy,and its related effects on tumor microenvironment(TME)in patients with advanced non-small cell lung cancer(NSCLC).Methods:A total of 30 patients with advanced NSCLC who were admitted to the Affiliated Hospital of Qingdao University from November 2021 to August 2023 were selected,and the expression levels of metabolites in feces of patients with different efficacy were detected by gas chromatography-mass spectrometry.Mouse CD8^(+)T cells were extracted and co-cultured with mouse lung cancer cells(LLC),and co-treated with sodium butyrate,PD-1 inhibitor monotherapy,and PD-1inhibitor combined with butyrate.The differences in cell proliferation,apoptosis,invasion and migration were detected by cell live death staining,CCK8 assay,Transwell assays and Wound Healing assay,respectively.ROS flow cytometry,Macrophage transformation assay and tubule formation assay were used to detect the H_(2)O_(2)production ability of cells and the effect on macrophage and vascular formation in TME.In vivo experiments and flow cytometry were used to detect the effect of butyrate on the tumor immune microenvironment in mouse metastases.Results:The content of butyrate in the feces of patients in the PD-1 inhibitor treatment group was significantly increased(P<0.05).After co-treatment with sodium butyrate combined with PD-1 inhibitor,the cytotoxicity of CD8^(+)T cells was significantly increased,the proliferation,invasion and migration ability of LLC cells were significantly decreased,and the apoptosis ability was significantly increased(P<0.05).The H_(2)O_(2)production capacity of LLC was significantly increased(P<0.05),the proportion of M1 macrophages was significantly increased(P<0.05),and the number of tubules formed was significantly decreased(P<0.05).The proportion of CD8^(+)T cells with metastases in mice increased significantly(P<0.05).Conclusion:Butyrate was highly expressed in patients in the PD-1 inhibitor response group for advanced non-sm

关 键 词：晚期非小细胞肺癌 短链脂肪酸 丁酸钠 PD-1抑制剂 肿瘤微环境 

分 类 号：R3[医药卫生—基础医学] R734.2