摘要
目的:探讨痛泻要方对慢性应激下小鼠大肠癌免疫微环境的影响及可能的作用机制。方法:取SPF级BABL/C雄性小鼠40只,随机分成正常组、应激组、痛泻要方组(13.65 g·kg^(-1))、痛泻要方应激组(13.65 g·kg^(-1)),每组10只。采用慢性束缚应激方法构建应激性小鼠模型,应激7 d后治疗组给予痛泻要方灌胃,在第14天通过强迫游泳实验和悬尾实验检测痛泻要方对应激后小鼠的行为学改变情况,同时对各组小鼠进行大肠癌皮下瘤种植。观察痛泻要方对小鼠体质量、肿瘤体积的影响;末次给药后,收集小鼠血清、瘤体样本。用免疫组化、流式细胞术检测瘤体内T淋巴细胞亚群(CD3^(+)、CD4^(+)、CD8^(+)及CD4^(+)/CD8^(+))含量变化;用酶联免疫吸附测定法(ELISA)检测小鼠外周血中皮质酮(CORT)、血清白细胞介素(IL)-2、γ干扰素(IFN-γ)、IL-6和IL-10含量;并用蛋白免疫印迹法(Western blot)检测小鼠核转录因子-κB(NF-κB)抑制蛋白(IκB)激酶α/β(IKKα/β)、NF-κB抑制蛋白α(IκBα)、NF-κB p65、磷酸化(p)-NF-κB p65蛋白表达。结果:与正常组比较,应激组小鼠瘤体体积增大(P<0.05);CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)数量下降(P<0.05,P<0.01);CD8^(+)数量有上升趋势;辅助性T细胞1(Th1)细胞因子IFN-γ含量降低(P<0.05);辅助性T细胞2(Th2)细胞因子IL-10含量升高(P<0.05);CORT含量升高(P<0.05);p-NF-κB p65、NF-κB p65蛋白表达升高,但差异无统计学意义;IKKα/β蛋白表达升高(P<0.05);IκBα蛋白表达明显下降(P<0.05)。与正常组比较,痛泻要方组瘤体生长速度下降,但差异无统计学意义;CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)数量显著上升(P<0.01);CD8^(+)数量下降(P<0.05);Th1细胞因子IL-2、IFN-γ含量升高(P<0.05);Th2细胞因子IL-6、IL-10含量降低(P<0.05);CORT含量降低,但差异无统计学意义;p-NF-κB p65蛋白表达下降,但差异无统计学意义;NF-κB p65、IKKα/β蛋白表达明显下降(P<0.05);IκBα蛋白表达显著
Objective:To explore the effect of Tongxie Yaofang on the immune microenvironment of colorectal cancer in mice under chronic stress and the underlying mechanism.Method:A total of 40 male SPF BABL/C mice were randomized into normal group,stress group,Tongxie Yaofang group(13.65 g·kg^(-1)),and Tongxie Yaofang-stress group(13.65 g·kg^(-1)),with 10 in each group.Chronic restraint stress was induced in mice and administration(ig) of Tongxie Yaofang began after 7 days of stress.On the 14th day,forced swim and tail suspension tests were used to examine the behavioral changes of mice after stress and the subcutaneous colorectal tumor was implanted in each group of mice.The effect of this prescription on the body mass and tumor volume of mice was observed.After the last administration,mouse serum and tumor samples were collected.The content of T lymphocytes(CD3^(+),CD4^(+),CD8^(+),and CD4^(+)/CD8^(+)) in tumor was detected by immunohistochemistry and flow cytometry and levels of corticosterone(CORT) in peripheral blood,and interleukin(IL)-2,interferon-γ(IFN-γ),IL-6,and IL-10 in the serum were determined by enzyme-linked immunosorbent assay(ELISA).The protein expression of inhibitor of nuclear factor-κB(IκB) kinase α/β(IKKα/β),nuclear factor-κB(NF-κB)α(IκBα),NF-κB p65,and phosphorylated(p)-NF-κB p65 was measured by Western blot.Result:Compared with the normal group,the stress group had large tumor volume(P<0.05),low content of CD3^(+),CD4^(+),and CD4^(+)/CD8^(+)(P<0.05,P<0.01),high content of CD8^(+),low content of T helper 1(Th1)-secreted IFN-γ(P<0.05),high content of T helper 2(Th2)-secreted IL-10(P<0.05) and CORT(P<0.05),high protein expression of p-NF-κB p65,NF-κB p65,and IKKα/β(P<0.05),and low protein expression of IκBα(P<0.05).Compared with the normal group,the Tongxie Yaofang group showed slow tumor growth,high content of CD3^(+),CD4^(+),and CD4^(+)/CD8^(+)(P<0.01),low content of CD8^(+)(P<0.05),high content of Th1-secreted IL-2 and IFN-γ(P<0.05),low content of Th2-secreted IL-6 and IL-10(P<
作者
杨懿
胡艳娥
蒋义芳
陈宁宁
严然
祝捷
由凤鸣
YANG Yi;HU Yane;JIANG Yifang;CHEN Ningning;YAN Ran;ZHU Jie;YOU Fengming(Traditional Chinese Medicine(TCM)Regulating Metabolic Diseases Key Laboratory of Sichuan Province,Hospital of Chengdu University of TCM,Chengdu 610075,China;Cancer Institute,Chengdu University of TCM,Chengdu 610072,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第12期46-54,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(82074315)
四川省科技厅项目(2021043)。
