清肺解毒汤对小鼠Lewis肺癌皮下移植瘤生长及肿瘤微环境免疫抑制的影响  

Effects of Qingfei Jiedu Decoction on the growth of subcutaneous grafts of Lewis lung cancer in mice and immunosuppression of tumor microenvironment

作　　者：潘俊杰 陈芳[2] 杨宏宽 汤军[2] 管彩虹[4] 石亚杰[4] 楼雅芳[4] PAN Jun-jie;CHEN Fang;YANG Hong-kuan;TANG Jun;GUAN Cai-hong;SHI Ya-jie;LOU Ya-fang(Dingqiao Hospital of Hangzhou/Dingqiao District of Hangzhou Hospital of Traditional Chinese Medicine,Hangzhou 310021,China;Zhejiang Provincial Hospital of Traditional Chinese Medicine,Hangzhou 310006,China;The People’s Hospital of Gaozhou,Maoming 525200,China;Hangzhou Hospital of Traditional Chinese Medicine,Hangzhou 310007,China)

机构地区：[1]杭州市丁桥医院/杭州市中医院丁桥院区,杭州310021 [2]浙江省中医院,杭州310006 [3]高州市人民医院,茂名525200 [4]杭州市中医院,杭州310007

出　　处：《中华中医药杂志》2020年第11期5752-5755,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：浙江省自然科学基金青年基金项目(No.LQ20H290004);浙江省中医药科技计划项目优秀青年人才基金(No.2020ZQ039)。

摘　　要：目的:研究清肺解毒汤对Lewis肺癌生长及肿瘤微环境免疫抑制的影响。方法:C57BL/6小鼠右腋皮下接种Lewis细胞建立肺癌模型,分Normal组,Model组,清肺解毒汤高、中、低剂量(Hd、Md、Ld)组和程序性细胞死亡蛋白1(PD-1)-程序性细胞死亡配体1(PD-L1)抑制剂(BMS-202)组,每组10只。Western blot检测瘤组织PD-L1;免疫荧光观察瘤组织Foxp3+Tregs密度;FCM检测脾脏CD4^+T、CD8^+T、CD4^+CD(25)^+Foxp3^+T百分比;ELISA测外周血白细胞介素(IL)-6、IL-2、干扰素-γ(INF-γ)。结果:Hd组、Md组、Ld组和BMS-202组抑瘤率为68.77%、60.91%、40.96%、55.24%。各组瘤重均小于Model组(P<0.05,P<0.01)。Foxp3^+Tregs密度:Hd组<BMS-202组<Md组<Ld组<Model组。与Model组比较,各组PD-L1均降低(P<0.05,P<0.01),且Hd组、Md组、Ld组显著低于BMS-202组(P<0.01),CD4^+T%均降低(P<0.01),Hd组、Md组和BMS-202组CD8^+T%升高且各组CD4^+CD(25)^+Foxp3^+T降低(P<0.05,P<0.01),各组IL-6降低,INF-γ升高,除Ld组外IL-2均升高。结论:清肺解毒汤通过减少肿瘤微环境Foxp3^+Tregs浸润和PD-L1表达,打破免疫抑制,提高CD8^+T比例和活性,发挥抗Lewis肺癌作用。Objective:To study the effects of Qingfei Jiedu(QFJD)Decoction on the growth of subcutaneous grafts of Lewis lung cancer in mice and immunosuppression of tumor microenvironment.Methods:Lewis cells were subcutaneously inoculated into the right axil of C57 BL/6 mice to establish lung cancer models,which were divided into normal group,model group,high-dose,medium-dose and low-dose of QFJD group(Hd,Md,Ld)and PD-1-PD-L1 inhibitor group(BMS-202),with 10 mice in each group.The PD-L1 in tumor tissues was detected by Western blot.The number of Foxp3^+Tregs in tumor tissues was observed by immunofluorescence laser confocal microscope.The proportion of CD4^+T,CD8^+T and CD4^+CD25^+Foxp3^+T in spleen was detected by flow cytometry.The concentrations of IL-6,IL-2 and INF-γin peripheral blood were detected by ELISA.Results:The tumor inhibition rate of Hd,Md,Ld and BMS-202 group were 68.77%,60.91%,40.96%and 55.24%,respectively.Tumor weight in each group was lower than model group(P<0.05,P<0.01).The number of Foxp3^+Tregs under laser scanning confocal microscope:Hd<BMS-202<Md<Ld<Model.Compared with model group,PD-L1 in each group was decreased(P<0.05,P<0.01)and Hd,Md and Ld group were significantly lower than BMS-202 group(P<0.01),CD4^+T%decreased(P<0.01),Hd,Md and BMS-202 group CD8^+T%increased and CD4^+CD25^+Foxp3^+T%decreased in each group(P<0.05,P<0.01),IL-6 was decreased in each group,INF-γwas significantly increased and IL-2 was increased in all groups except Ld group.Conclusion:QFJD Decoction plays an antiLewis lung cancer role by reducing the infiltration of Foxp3^+Tregs and the expression of PD-L1 in the tumor microenvironment,breaking the immunosuppression,increasing the proportion and activity of CD8^+T.

关 键 词：LEWIS肺癌 清肺解毒汤 肿瘤微环境 免疫抑制 T淋巴细胞 细胞因子 程序性细胞死亡配体1 

分 类 号：R285.5[医药卫生—中药学]