丹参总酚酸联合anti-PD-L1调控髓源性巨噬细胞浸润抑制乳腺癌发生发展  

Total salvianolic acid combined with anti-PD-L1 inhibits development of breast cancer by regulating infiltration of myeloid macrophages

作　　者：宋梦瑶 钱程 陆茵[1,2] SONG Meng-yao;QIAN Cheng;LU Yin(Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica,School of Pharmacy;Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine(TCM)Prevention and Treatment of Tumor,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学药学院,江苏省中药药效与安全性评价重点实验室 [2]江苏省中医药防治肿瘤协同创新中心,江苏南京210023

出　　处：《中国药理学通报》2023年第10期1884-1890,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81973734);江苏省研究生创新项目(No KYCX21-1794)。

摘　　要：目的探究丹参总酚酸(total salvianolic acid,TSA)联合anti-PD-L1通过调控髓源性巨噬细胞瘤内浸润抑制乳腺癌的发生发展。方法构建E0771乳腺癌皮下肿瘤模型。25只小鼠随机分为空白组、模型组、TSA组(TSA 10 g·kg^(-1)),anti-PD-L1组(anti-PD-L110 mg·kg^(-1)),TSA联合anti-PD-L1组(TSA 10 g·kg^(-1)+anti-PD-L110 mg·kg^(-1))。TSA组每日灌胃给药,anti-PD-L1每3天1次腹腔注射,连续给药14 d。记录小鼠肿瘤体积变化及肿瘤、肝脏及脾脏重量。ELISA检测小鼠血浆中IL-6,MCP-1的含量,qPCR检测cxcl1,cxcl2,cxcl3,ccl2,gm-csf mRNA的表达,流式、免疫组化检测小鼠肿瘤、淋巴中髓源性巨噬细胞浸润情况,流式检测淋巴、脾脏CD4+T细胞、CD8+T细胞数量变化。结果与模型组相比,TSA,anti-PD-L1,TSA联合anti-PD-L1均可抑制E0771乳腺癌生长,减少IL-6、MCP-1分泌,降低cxcl1,cxcl2,cxcl3,ccl2,gm-csf mRNA表达,抑制髓源性巨噬细胞向肿瘤的募集,增加淋巴及脾脏内CD4+T细胞、CD8+T细胞数量,TSA联合anti-PD-L1治疗效果更佳显著。结论TSA联合anti-PD-L1显著抑制E0771乳腺癌皮下瘤发生发展进程,可能通过抑制髓源性巨噬细胞向肿瘤浸润,增强机体免疫效应发挥作用。Aim To explore the effect of total salvianolic acid(TSA)combined with anti-PD-L1 on the development of breast cancer by inhibiting the infiltration of myelogenous macrophages into tumors.Methods E0771 breast cancer subcutaneous tumor model was constructed.Twenty-five mice were randomly divided into control group,model group,total salvianolic acid group(TSA 10 g·kg^(-1))and total salvianolic acid combined with anti-PD-L1 group(TSA 10 g·kg^(-1)+anti PD-L110 mg·kg^(-1)).The total salvianolic acid group was administered daily by gavage,and anti-PD-L1 was injected intraperitoneally,once every three days,lasting for 14 days.Tumor volume,tumor weight,liver and spleen index were recorded.The content of IL-6 and MCP-1 in plasma was detected by ELISA.The mRNA level of cxcl1,cxcl2,cxcl3,ccl2,and gm-csf was detected by qPCR.The infiltration of myelogenous macrophages in tumor and lymph was detected by flow cytometry and immunohistochemistry,and the changes of CD4+T cells,CD8+T cells in lymph and spleen were detected by flow cytometry.Results Compared with the model group,total salvianolic acid,anti-PD-L1,total salvianolic acid combined with anti-PD-L1 inhibited the growth of E0771 breast cancer,reduced the secretion of IL-6 and MCP-1 and the expression of cxcl1,cxcl2,cxcl3,ccl2,gm-csf mRNA level,suppressed the recruitment of myelogenous macrophages into tumors and increased the number of CD4+T cells,CD8+T cells in lymph and spleen.The effect of total salvianolic acid combined with anti-PD-L1 was superior.Conclusions Total salvianolic acid combined with anti-PD-L1 significantly inhibits the progression of subcutaneous tumor of E0771 breast cancer by enhancing the immune effect and inhibiting the infiltration of myeloid macrophages into the tumor.

关 键 词：丹参总酚酸 乳腺癌 anti-PD-L1 髓源性巨噬细胞 免疫疗法 中药现代化 

分 类 号：R-332[医药卫生] R282.71R329.24R392.12R737.9