IL-33介导巨噬细胞极化影响非小细胞肺癌患者Tre g/Th17失衡  

IL-33 Mediates Tumor-associated Macrophages to Influence Treg/Th17 Imbalance in the Tumor Microenvironment of Non-small Cell Lung Cancer

作　　者：谭希 刘慧芳[2] 王晶[2] 胡婷婷 崔萌萌 姜敏[2] TAN Xi;LIU Huifang;WANG Jing;HU Tingting;CUI Mengmeng;JIANG Min(Fourth Clinical Medical College of Xinjiang Medical University,Urumqi 830000,China;The Fourth Affiliated Hospital of Xinjiang Medical University,Xinjiang Key Laboratory of Respiratory Disease Research,Urumqi 830000,China)

机构地区：[1]新疆医科大学第四临床医学院,乌鲁木齐830000 [2]新疆医科大学第四附属医院,新疆呼吸病研究重点实验室,乌鲁木齐830000

出　　处：《宁夏医科大学学报》2024年第9期878-885,共8页Journal of Ningxia Medical University

基　　金：国家自然科学基金项目(82360866);新疆维吾尔自治区重点实验室开放课题(2020D04030)。

摘　　要：目的 探讨非小细胞肺癌(NSCLC)中白细胞介素(IL)-33介导巨噬细胞极化对调节性T细胞(Treg)/辅助性T细胞17(Th17)失衡的影响。方法 选择2020年9月至2021年8月新疆医科大学第四附属医院收治的42例NSCLC住院患者(NSCLC组)及40例健康体检者(对照组)作为研究对象。采用流式细胞术检测两组研究对象外周血中Th17、Treg及巨噬细胞/单核细胞的数量;使用磁珠法分选NSCLC组外周血单个核细胞和对照组外周血CD4^(+)T细胞,并使用IL-33诱导巨噬细胞极化,将NSCLC组IL-33极化后的巨噬细胞与对照组CD4^(+)T细胞共培养;RT-qPCR法检测IL-10、肿瘤坏死因子-α(TNF-α)、IL-1β、转化生长因子-β(TGF-β)mRNA水平及共培养后维甲酸相关孤核受体γt(RORγt)、叉头蛋白P3(FOXP3)mRNA水平;CBA多因子试剂盒检测细胞培养上清液中TNF-α、IL-1β、IL-10、TGF-β的含量。结果 外周血中,NSCLC组CD14^(+)CD68^(+)CD86^(+)CD206-样M1、CD14^(+)CD68^(+)CD86-CD206^(+)样M2巨噬细胞比例及CD14^(+)CD68^(+)CD86-CD206^(+)样M2/CD14^(+)CD68^(+)CD86^(+)CD206-样M1均高于对照组(P均<0.05),NSCLC组Treg细胞水平及Treg/Th17比例均高于对照组(P均<0.05);IL-33诱导的巨噬细胞培养上清液中IL-10 mRNA、TGF-β mRNA相对表达水平均增加(P均<0.05);IL-33诱导的巨噬细胞与CD4^(+)T细胞共培养后,FOXP3 mRNA水平高于与M0、M1共培养组(P<0.05)。结论 IL-33能够促进NSCLC中巨噬细胞M2极化,M2巨噬细胞促进Treg/Th17比例增加,上调Treg特异性转录因子FOXP3,可能与其促进IL-10、TGF-β分泌增多有关。Objective To investigate the effect of interleukin(IL)-33 mediated macrophage polarization on the imbalance of regulatory T cells(Treg)/helper T cell-17(Th17)in non-small cell lung cancer(NSCLC).Methods The study enrolled 42 NSCLC inpatients(NSCLC group),40 healthy subjects(control group)admitted to the Fourth Clinical Medical College of Xinjiang Medical University from September 2020 to August 2021 as the study subjects.Th17,Treg and macrophages/monocytes in peripheral blood of the two groups were detected by flow cytometry.Magnetic beads were used to separate peripheral blood mononuclear cells from the NSCLC group and CD4^(+)T cells from the control group,and IL-33 was used to induce macrophage polarization.The NSCLC macrophages treated with or without IL-33 were co-cultured with CD4^(+)T cells.The mRNA levels of IL-10,TNF-α,IL-1β,TGF-β,RORγt and FOXP3 were detected by RT-qPCR.Contents of TNF-α,IL-1β,IL-10 and TGF-βin supernatant of cell culture were detected by CBA multi-factor kit.Results The levels of CD14^(+)CD68^(+)CD86^(+)CD206-M1 and CD14^(+)CD68^(+)CD86-CD206^(+)M2 macrophages and the ratio of CD14^(+)CD68^(+)CD86-CD206^(+)M2/CD14^(+)CD68^(+)CD86^(+)CD206-M1 in peripheral blood of NSCLC group were higher than those in the control group(P all<0.05).In addition,the level of Treg and the ratio of Treg/Th17 in NSCLC group were higher than those in the control group(P all<0.05).The concentration of IL-10 mRNA and TGF-βmRNA increased in the supernatant of macrophages induced by IL-33(P all<0.05).FOXP3 mRNA level of M2 type macrophages co-cultured with CD4^(+)T cells was significantly higher than that of M0 and M1 co-cultured groups(P<0.05).Conclusion IL-33 can promote M2 polarization of macrophages in NSCLC,and M2 macrophages promote an increase in Treg/Th17 ratio,upregulating the Treg specific transcription factor FOXP3.This mechanism may be related to its promotion of increased IL-10 and TGF-βsecretion.

关 键 词：非小细胞肺癌 巨噬细胞 白细胞介素-33 调节性T细胞 辅助性T细胞17 

分 类 号：R734.2[医药卫生—肿瘤]