天门冬汤敷贴通过IL-1β/STAT3信号通路调控髓源抑制性细胞抑制小鼠肺癌的机制研究  

Mechanism of Tianmendong Decoction(天门冬汤)Application Inhibiting Lung Cancer in Mice by Regulating Myeloid Inhibitory Cells

作　　者：梁馨月 苏琳 李伟 朱杨壮壮 阎学伟 祝晓雯[1] 胥孜杭 邹纯朴[1] LIANG Xinyue;SU Lin;LI Wei;ZHU-YANG Zhuangzhuang;YAN Xuewei;ZHU Xiaowen;XU Zihang;ZOU Chunpu(School of Traditional Chinese Medicine,Shanghai University of Chinese Medicine,Shanghai 201203,China;Shanghai Skin Disease Hospital Aesthetic Medicine,Shanghai 200050,China)

机构地区：[1]上海中医药大学中医学院,上海201203 [2]上海市皮肤病医院,上海200050

出　　处：《辽宁中医杂志》2025年第3期194-198,F0003,共6页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82274229);上海市青年科技启明星项目(23QA1409100);上海中医药大学杏林青年学者人才计划项目(B1-GY21-409-04-37)。

摘　　要：目的研究天门冬汤敷贴(以下均称TD敷贴)对肺癌原位模型小鼠的治疗作用及作用机制。方法依据课题组前期构建的小鼠肺癌原位模型方法,在小鼠左侧肺叶内注射荧光素酶标记的小鼠Lewis肺癌细胞(Lewis lung carcinoma,LLC-luc)建立小鼠肺癌原位模型。将造模小鼠随机分为对照组、TD敷贴组;分别在小鼠背部以肺俞穴为中心,0.5 cm×0.5 cm为范围贴敷敷贴,对照组贴敷空白敷贴,TD敷贴组贴敷含药敷贴,每日更换敷贴,连续治疗25 d。观察小鼠体征,记录小鼠体质量及生存期。流式细胞术检测小鼠肿瘤内主要免疫细胞的变化。通过网络药理学预测天门冬汤药物作用于髓源抑制性细胞(myeloid-derived suppressor cells,MDSCs)的潜在靶点。实时荧光定量聚合酶链式反应(Reverse Transcription-Polymerase Chain Reaction,RT-qPCR)检测小鼠白细胞介素-1β(interleukin-1β,IL-1β)、信号转导和转录活化因子3(signal transduction and transcription activator 3,STAT3)、精氨酸酶(arginase 1,Arg1)、一氧化氮合酶(inducible nitric oxide synthase,iNOS)、S100钙结合蛋白A9(S100 calcium binding protein A9,S100A9)基因表达。蛋白免疫印迹(Western blot)法检测小鼠Arg1、iNOS、S100A9、STAT3、p-STAT3蛋白表达。结果TD敷贴治疗可延长肺癌小鼠生存期(P<0.05);TD敷贴对小鼠体质量无直接影响(P>0.05);TD敷贴治疗后肺癌小鼠肿瘤中MDSCs显著降低(P<0.001),T细胞数量增加(P<0.05),其余免疫细胞如自然杀伤细胞、巨噬细胞、树突状细胞、调节性T细胞无明显变化(P>0.05)。网络药理学预测天门冬汤药物靶点通过IL-1β靶向MDSCs发挥作用。TD敷贴治疗后与MDSCs活化相关的Arg1、iNOS、S100A9 mRNA及蛋白表达水平显著降低(P<0.01);IL-1β、STAT3 mRNA表达降低(P<0.01);STAT3、p-STAT3及MDSCs活化相关的Arg1、iNOS、S100A9蛋白表达下调(P<0.05)。结论天门冬汤敷贴通过IL-1β/STAT3信号通路抑制MDSCs的增殖与活化从而延长肺癌Objective To study the therapeutic effect and mechanism of Tianmendong Decoction(天门冬汤)application(hereinafter referred to as TD application)on in-situ lung cancer model mice.Methods According to the mouse lung cancer in situ model established by the research group,the mouse lung cancer in situ model was established by injecting luciferase labeled mouse Lewis lung cancer cells(LLC-luc)into the left lung lobe of mice.The mice were randomly divided into control group and TD application group.On the back of mice,with Feishu(BL13)as the center,0.5 cm×0.5 cm was applied.The blank application was applied in the control group,and drug-containing application was applied in the TD application group,and the application was changed every day for 25 consecutive days.The appearance of the mice was observed,and the weight and survival time of the mice were recorded.The changes of major immune cells in mouse tumors were detected by flow cytometry.Network pharmacology was used to predict the potential target of the action of Tianmendong Decoction on myeloid inhibitory cells(MDSCs).The expression of interleukin-1β(IL-1β),signal transduction and transcriptional activating factor 3(STAT3),arginase(Arg1),nitric oxide synthase(iNOS)and S100 calcium binding protein A9(S100A9)were detected by Reverse Transcription-Polymerase Chain Reaction(RT-qPCR).The expressions of Arg1,iNOS,S100A9,STAT3 and p-STAT3 were detected by Western blot.Results TD application could prolong the survival time of lung cancer mice(P<0.05).TD application had no direct effect on body weight of mice(P>0.05).After TD application,the MDSCs in lung cancer mice were significantly decreased(P<0.001),and the number of T cells was increased(P<0.05).Macrophages,DCs and Treg cells had no significant changes(P>0.05).Network pharmacology predicted that the drug target of Tianmendong Decoction worked by targeting MDSCs with IL-1β.The mRNA and protein expression levels of Arg1,iNOS and S100A9 associated with MDSCs activation were significantly decreased after TD appl

关 键 词：天门冬汤 敷贴 髓源抑制性细胞 肺癌原位模型 肿瘤外治 

分 类 号：R285[医药卫生—中药学]