灸药联合对乳腺癌荷瘤小鼠肿瘤组织免疫检查点的影响  

Effect of moxibustion combined with chemotherapy on immune checkpoints in tumor tissue of breast cancer-bearing mice

作　　者：张飞程 高田宇 张晨曦 李继娟 梁新月 张晓琪 贾春生 潘丽佳 ZHANG Fei-cheng;GAO Tian-yu;ZHANG Chen-xi;LI Ji-juan;LIANG Xin-yue;ZHANG Xiao-qi;JIA Chun-sheng;PAN Li-jia(College of Acupuncture-Moxibustion and Tuina,Hebei University of Chinese Medicine,Shijiazhuang 050200,China)

机构地区：[1]河北中医药大学针灸推拿学院,石家庄050200

出　　处：《针刺研究》2025年第3期319-326,共8页Acupuncture Research

基　　金：河北中医药大学2022年博士科研基金项目(No.BSZ2022006);河北省自然科学基金项目(No.H2023423029)。

摘　　要：目的:观察灸药联合对乳腺癌小鼠肿瘤组织中免疫检查点程序性死亡蛋白1(PD-1)、T细胞免疫球蛋白黏蛋白-3(TIM-3)、细胞毒性T淋巴细胞相关蛋白4(CTLA-4)的影响,探究艾灸联合化疗药对乳腺癌的干预作用及其机制。方法:将BALB/c雌性小鼠随机分为模型组、化疗组、艾灸组、灸药组,每组10只。在小鼠左侧第4乳头下脂肪垫接种4T1肿瘤细胞,建立乳腺癌荷瘤小鼠模型。化疗组腹腔注射阿霉素溶液2.5 mg/kg,每3 d注射1次;艾灸组给予双侧“足三里”艾炷灸,每穴每次2壮,每2 d治疗1次;灸药组同时给予DOX注射和艾炷灸干预;各组均干预21 d。每天记录小鼠体质量和肿瘤体积,治疗结束后处死取材,HE染色观查肿瘤组织病理形态,免疫组织化学法和Western blot法检测小鼠肿瘤组织中PD-1、TIM-3、CTLA-4的阳性表达和蛋白表达情况。结果:干预后与模型组比较,各治疗组乳腺癌荷瘤小鼠体质量均升高(P<0.01,P<0.05),肿瘤体积均减小(P<0.01),肿瘤组织病理形态上均出现不同程度的肿瘤细胞退变,肿瘤组织PD-1、TIM-3、CTLA-4的阳性表达和蛋白表达均降低(P<0.01,P<0.05)。与化疗组比较,艾灸组小鼠体质量升高(P<0.01);灸药组小鼠肿瘤体积减小(P<0.01);艾灸组和灸药组小鼠肿瘤组织PD-1、TIM-3、CTLA-4的阳性表达和蛋白表达均降低(P<0.01,P<0.05)。与艾灸组比较,灸药组小鼠体质量下降(P<0.01),肿瘤体积减小(P<0.01),肿瘤组织中肿瘤细胞退变程度更高,肿瘤组织PD-1、TIM-3、CTLA-4的阳性表达和蛋白表达均降低(P<0.01)。结论:艾灸联合化疗药在抑制乳腺癌小鼠肿瘤生长方面比单独艾灸、单独化疗治疗效果更加显著,能降低肿瘤组织中PD-1、TIM-3、CTLA-4的表达,提示降低免疫检查点的表达水平可能是艾灸联合化疗药治疗肿瘤的作用机制之一。Objective To observe the effects of moxibustion combined with chemotherapy on immune checkpoints including programmed cell death protein 1(PD-1),T cell immunoglobulin domain and mucin domain(TIM-3)and cytotoxic T-lymphocyte-associated protein 4(CTLA-4)in the tumor tissue of breast cancer-bearing mice,so as to explore the effect and mechanisms of moxibustion combined with chemotherapy on breast cancer.Methods Forty BALB/c female mice were randomly divided into model,chemotherapy,moxibustion and combination groups,with 10 mice in each group.4T1 tumor cells were inoculated into the fat pad under the left fourth nipple of the mice to establish the breast cancer-bearing mice model.The chemotherapy group was injected intraperitoneally with doxorubicin(DOX)solution at 2.5 mg/kg once every 3 days;the moxibustion group received bilateral“Zusanli”(ST36)moxibustion with 2 moxa cones per point,treated every 2 days;the combination group received both DOX injection and moxibustion intervention.The above interventions lasted for 21 days.The body weight and tumor volume of the mice were recorded daily.HE staining was used to observe tumor tissue pathological morphology.Immunohistochemistry and Western blot were used to detect the positive expression and protein expression levels of PD-1,TIM-3,and CTLA-4 in the tumor tissue.Results Compared with the model group after intervention,the body weight was increased(P<0.01,P<0.05),the tumor volume was decreased(P<0.01),the pathological morphology of tumor tissue showed varying degrees of tumor cells degeneration,and the positive expression and protein expression levels of PD-1,TIM-3,and CTLA-4 in tumor tissue were decreased(P<0.01,P<0.05)of mice in the chemotherapy,moxibustion and combination groups.Compared with the chemotherapy group,the body weight of mice in the moxibustion group was higher(P<0.01);the tumor volume of mice in the combination group was smaller(P<0.01);the positive expression and protein expression levels of PD-1,TIM-3,and CTLA-4 were further reduced(P<0.01,P<0.05

关 键 词：艾灸 乳腺癌 免疫检查点 程序性死亡蛋白1 T细胞免疫球蛋白黏蛋白-3 细胞毒性T淋巴细胞相关蛋白4 

分 类 号：R245[医药卫生—针灸推拿学]