乌梅丸激活STING促进CD8^(+)T细胞浸润活化抗结直肠癌的机制  

Mechanism of Wumei Pill activating STING to promote CD8^(+)T cell infiltration and activation against colorectal cancer

作　　者：支景琳 冯媛媛[1] 刘宁宁[1] 陈念芝 ZHI Jinglin;FENG Yuanyuan;LIU Ningning;CHEN Nianzhi(Department of Oncology,Shuguang Hospital Affiliated to 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肿瘤科,上海201203

出　　处：《上海中医药杂志》2025年第5期1-7,共7页Shanghai Journal of Traditional Chinese Medicine

基　　金：中国博士后科学基金项目(2023MD734135);重庆市博士后科学基金项目(CSTB2023NSCQ-BHX0019)。

摘　　要：目的探讨乌梅丸对结直肠癌的抗癌能力和对T细胞表达的影响及其作用机制。方法采用皮下移植瘤构建法建立小鼠结直肠癌模型,然后随机将小鼠分为对照组(0.9%氯化钠溶液),乌梅丸低剂量组(20g/kg)和乌梅丸高剂量组(40g/kg);记录小鼠体质量、生存期、肿瘤体积以及肿瘤质量。随后采用苏木精-伊红(HE)染色法观察乌梅丸对小鼠结直肠肿瘤的影响,采用免疫组织化学染色法检测乌梅丸对小鼠肿瘤组织中细胞核增殖抗原-67(Ki67)表达的影响,结合流式细胞术分析乌梅丸对结直肠癌小鼠肿瘤组织中CD3^(+)T细胞、CD4^(+)T细胞、CD8^(+)T细胞数量的影响,通过免疫荧光染色法观察乌梅丸对小鼠肿瘤组织中磷酸化干扰素刺激因子(p-STING)表达的影响,采用酶联免疫吸附分析(ELISA)法检测乌梅丸对小鼠血清中β干扰素(IFN-β)含量的影响,采用实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法检测乌梅丸对小鼠肿瘤组织中IFN-βmRNA表达的影响。结果与对照组比较,乌梅丸干预可降低肿瘤质量和肿瘤体积,同时延长结直肠癌小鼠的生存期(P<0.01)。HE染色和免疫组织化学染色法检测结果显示,与对照组相比,乌梅丸干预后肿瘤坏死部分明显增多,且肿瘤组织中Ki67表达降低(P<0.01)。通过流式细胞术结果分析得出,与对照组相比,乌梅丸对肿瘤组织中CD3^(+)T细胞和CD4^(+)T细胞的数量无明显影响(P>0.05),但却显著提高了CD8^(+)T细胞的数量(P<0.01)。同时,乌梅丸上调了结直肠癌小鼠肿瘤组织中p‐STING水平(P<0.01),以及上调了血清及肿瘤组织中IFN-β的表达水平(P<0.01)。结论乌梅丸可通过激活干扰素刺激因子(STING)信号通路促进肿瘤免疫微环境中CD8^(+)T细胞的浸润,从而发挥抗结直肠癌的作用。Objective To investigate the effects of Wumei Pill on the anti-cancer ability and T cell expression of colorectal cancer and its potential mechanism.Methods Colorectal cancer mice model were established by subcutaneous tumor transplantation method,and then randomly divided into control group(0.9%sodium chloride solution),Wumei Pill low-dose group(20 g/kg)and Wumei Pill high-dose group(40 g/kg).The mass,survival time,tumor volume and tumor mass of the mice were recorded.Subsequently,the effect of Wumei Pill on colorectal tumors in mice was observed by hematoxylin-eosin(HE)staining.The effect of Wumei Pill on nuclear proliferating antigen-67(Ki67)expression in mouse tumor tissue was detected by immunohistochemistry staining.Flow cytometry was used to analyze the effects of Wumei Pill on the number of CD3^(+)T cells,CD4^(+)T cells and CD8^(+)T cells in the tumor tissues of model mice.The effect of Wumei Pill on the expression of phosphorylated interferon stimulating factor(p-STING)in tumor tissue of mice was observed by immunofluorescence staining.The effect of Wumei Pill on serum interferon-β(IFN-β)content of mice was detected by enzyme-linked immunosorbent assay(ELISA).The effect of Wumei Pill on IFN-βmRNA expression in mouse tumor tissue was detected by real-time quantitative polymerase chain reaction(RT-qPCR).Results Wumei Pill decreased the mass and volume of tumor,and prolonged the survival time of colorectal cancer mice(P<0.01).HE staining and immunohistochemistry staining results showed that the necrotic part of tumor increased significantly after Wumei Pill intervention compared with the control group,and Ki67 levels increased in tumor tissues(P<0.01).Flow cytometry result showed that Wumei Pill had no significant effect on the number of CD3^(+)T cells and CD4^(+)T cells in tumor tissues(P>0.05),but increased the number of CD8^(+)T cells(P<0.01).Wumei Pill up-regulated the level of p-STING in tumor tissues of colorectal cancer mice(P<0.01),and up-regulated the expression level of IFN-βin serum and tumor

关 键 词：结直肠癌 乌梅丸 T淋巴细胞 干扰素刺激因子 作用机制 中药研究 

分 类 号：R285.5[医药卫生—中药学]