肺岩宁方对Lewis荷瘤小鼠肿瘤相关性巨噬细胞极化的调控作用研究  

Study on the Regulating Effect of Feiyanning Prescription (肺岩宁方) on the Polarization of Tumor-associated Macrophages in Lewis Tumor-bearing Mice

作　　者：张琦君[1,2] 邓海滨 车勇[2] 殷书敏 徐振晔 ZHANG Qi-jun;DENG Hai-bin;CHE Yong;YIN Shu-min;XU Zhen-ye(Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Jiangwan Hospital of Hongkou District of Shanghai,Shanghai 200434f China;Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学,上海201203 [2]上海市虹口区江湾医院,上海200434 [3]上海中医药大学附属龙华医院,上海200032

出　　处：《中医药导报》2021年第4期1-4,19,共5页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：上海市教育委员会科研创新计划项目(2017-01-07-00-10-E00064);上海市基层名老中医专家传承研究工作室(HGY-MZY-2018-04)。

摘　　要：目的:探究肺岩宁方对Lewis荷瘤小鼠肿瘤相关性巨噬细胞极化的调控作用及分子机制。方法:36只雄性C57BL/6小鼠随机分为正常对照组、模型对照组、正常肺岩宁组、模型肺岩宁组,采用Lewis肺癌细胞株皮下注射建立移植瘤模型。实验干预2周后测小鼠瘤质量计算抑瘤率,免疫组化染色后采用Image J图像分析软件计算小鼠皮下瘤组织中CD68、CD206阳性表达结果,ELISA检测小鼠血清中IL-4、IL-10含量,Western blotting检测小鼠皮下瘤组织中PI3K、C/EBPβ、PD-1、PD-L1表达水平。结果:模型肺岩宁组小鼠瘤质量明显低于模型对照组(P<0.05),模型肺岩宁组抑瘤率为60.59%。模型肺岩宁组小鼠瘤组织中CD68的平均光密度值明显高于模型对照组(P<0.05),CD206的平均光密度值明显低于模型对照组(P<0.05)。模型对照组小鼠血清IL-4、IL-10含量明显高于正常对照组、正常肺岩宁组(P<0.05),模型肺岩宁组小鼠血清IL-4、IL-10含量明显低于模型对照组(P<0.05)。模型肺岩宁组小鼠瘤组织中PI3K、C/EBPβ、PD-1、PD-L1表达水平明显低于模型对照组(P<0.05)。结论:肺岩宁方可能通过下调IL-4、IL-10分泌,抑制PI3K/C/EBPβ和PD-1/PD-L1信号通路,调控肿瘤微环境中TAMs极化,促使其表型向M1型转变,从而发挥抑瘤作用。Objective: To explore the regulatory effect and molecular mechanism of Feiyanning Prescription on the polarization of tumor-associated macrophages in Lewis tumor-bearing mice. Methods: A total of 36 male C57 BL/6 mice were randomly divided into normal control group, normal Feiyanning group, model control group and model Feiyanning group. Transplanted tumor model was established by subcutaneous injection of Lewis lung cancer cell line. After 2 weeks of experimental intervention, the tumor inhibition rate was calculated. IHC and ImageJ soft were used to observe the expression of CD68, CD206. ELISA was used to observe the concentration of IL-4 and IL-10 in serum. Western blotting was used to observe the expression levels of PI3 K, C/EBPβ, PD-1 and PD-L1 in subcutaneous tumor tissue. Results: The tumor quality of the model Feiyanning group was significantly lower than that of the model control group(P<0.05), and the tumor inhibition rate was 60.59%. The average optical density value of CD68 in tumor tissue of model Feiyanning group was significantly higher than that of model control group(P<0.05), and the average optical density value of CD206 was significantly lower than that of model control group(P<0.05). The serum levels of IL-4 and IL-10 in the model control group were significantly higher than those in the normal control group and the normal Feiyanning group(P<0.05). The serum levels of IL-4 and IL-10 in the model Feiyanning group were significantly lower than those in the model control group(P<0.05).The expression levels of PI3K, C/EBPβ, PD-1 and PD-L1 in the tumor tissues of the model Feiyanning group were significantly lower than that of the model control group(P<0.05). Conclusion: Feiyanning Prescription may regulate the polarization of TAMs in tumor microenvironment by inhibiting the secretion of IL-4 and IL-10 and PI3 K/C/EBPβ and PD-1/PD-L1 signal pathway, so as to promote the phenotype transformation to M1 type,thereby exerting a tumor suppressor effect.

关 键 词：肺癌 肿瘤免疫微环境 肺岩宁方 肿瘤相关性巨噬细胞 小鼠 

分 类 号：R285.5[医药卫生—中药学]