补肾解毒方抑制肿瘤相关巨噬细胞激活介导的大肠癌转移的机制研究  

Study on the mechanism of Bushen Jiedu Formula on inhibiting the metastasis of colorectal cancer mediated by tumor-associated macrophages activation

作　　者：刘静雯 王瑜 贾茹 冯媛媛[1] 孙筱婷 王炎[1] 李琦[1,2] LIU Jing-wen;WANG Yu;JIA Ru;FENG Yuan-yuan;SUN Xiao-ting;WANG Yan;LI Qi(Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Academy of Integrative Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院,上海201203 [2]上海中医药大学中西医结合研究院,上海201203

出　　处：《中华中医药杂志》2023年第3期1231-1235,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82030118,No.82122075,No.82104619)。

摘　　要：目的:探讨补肾解毒方(BSJDR)调控肿瘤相关巨噬细胞(TAMs)极化对大肠癌细胞迁移侵袭的影响。方法:体外培养小鼠单核巨噬细胞RAW264.7,分别使用PBS、白细胞介素(IL)-4、IL-4+BSJDR低剂量(-L)、IL-4+BSJDR中剂量(-M)以及IL-4+BSJDR高剂量(-H)刺激RAW264.748 h。应用流式细胞术检测TAMs分群比及荧光定量PCR法(qPCR)检测M2型巨噬细胞精氨酸酶(Arg1)、甘露糖受体(Cd206)、Cd163 mRNA表达。收集条件培养基(CM)构建微环境培养体系,应用Transwell与划痕实验观察BSJDR对MC38细胞侵袭迁移的影响。结果:流式细胞术显示,IL-4组M2巨噬细胞分群比为77.7%,显著高于Control组的1.01%、IL-4+BSJDR-L组的40.1%、IL-4+BSJDR-M组的31.4%以及IL-4+BSJDR-H组的29.8%(P<0.01)。qPCR结果显示,IL-4组Arg1、Cd206、Cd163 mRNA表达较Control组显著升高(P<0.01),IL-4+BSJDR组Arg1、Cd206、Cd163 mRNA表达均较IL-4组显著降低(P<0.01,P<0.05)。Transwell及划痕实验显示,IL-4-CM组中MC38细胞侵袭迁移能力增强(P<0.05,P<0.01);(IL-4+BSJDR)-CM组MC38细胞侵袭迁移能力较IL-4-CM组显著降低(P<0.01)。结论:BSJDR可通过抑制TAMs向M2型巨噬细胞极化抑制大肠癌转移。Objective:To explore the Bushen Jiedu Formula(BSJDF)on regulating tumor-associated macrophages(TAMs)polarization and further reveal its inhibition mechanisms on invasion and metastasis of colorectal cancer(CRC).Methods:Mouse monocyte-macrophage RAW264.7 cultured in vitro was stimulated with PBS,IL-4,IL-4+BSJDF-L,IL-4+BSJDF-M and IL-4+BSJDF-H for 48 h.Flow cytometry was used to analyze the TAMs ratio and real-time quantitative PCR(qPCR)was used to detect the M2-TAMs mRNA expressions of Arg-1,Cd206 and Cd163.Transwell and scratch experiments to observe the effect of BSJDF on the invasion and migration of MC38 by constructing a microenvironmental culture system by collecting conditioned medium.Results:Flow cytometry showed that the subgroup ratio of M2-TAMs in the IL-4 group was 77.7%,significantly higher than 1.01%in the control group,40.1%in IL-4+BSJDF-L group,31.4%in IL-4+BSJDF-M group and 29.8%in IL-4+BSJDF-H group(P<0.01).qPCR showed that the expression of M2-TAMs mRNA of the IL-4 group was significantly higher than that the control group(P<0.01);the expression of M2-TAMs mRNA in the IL-4+BSJDF group was decreased than the IL-4 group(P<0.01,P<0.05).Transwell assay and scratch experiments showed that IL-4-CM group significantly enhanced the invasion of MC38 cell(P<0.05,P<0.01),the(IL-4+BSJDF)-CM group significantly lower than the IL-4-CM group(P<0.01).Conclusion:BSJDF inhibits CRC metastasis by inhibiting the activation of M2-TAMs in the tumor microenvironment.

关 键 词：补肾解毒方 肿瘤相关巨噬细胞 M2型巨噬细胞 大肠癌 侵袭转移 

分 类 号：R285.5[医药卫生—中药学]