黄芩汤对结肠炎相关性结肠癌肠道炎症和增殖的干预机制  

Intervention Mechanism of Huangqintang on Intestinal Inflammation and Proliferation in Colitis-associated Colon Cancer

作　　者：朱琳[1,2] 王敦方 冯雪 张彩娟 刘海帆 刘雅清 刘滨 刘丽[2] 杨伟鹏 ZHU Lin;WANG Dunfang;FENG Xue;ZHANG Caijuan;LIU Haifan;LIU Yaqing;LIU Bin;LIU Li;YANG Weipeng(Heilongjiang University of Chinese Medicine,Harbin 150040,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]黑龙江中医药大学,哈尔滨150040 [2]中国中医科学院中药研究所,北京100700

出　　处：《中国实验方剂学杂志》2023年第22期1-10,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82074328,81473592);中国中医科学院科技创新工程项目(CI2021A04604);中国中医科学院中药研究所自选课题-人才引进项目(ZXKT20033)。

摘　　要：目的:研究黄芩汤对结肠炎相关性结肠癌(CAC)小鼠模型的药效,并探究黄芩汤在CAC中调节免疫功能和炎症反应,抑制细胞异常增殖,延缓或抑制CAC形成的机制。方法:C57BL/6J小鼠按体质量随机分为正常组、模型组、美沙拉嗪组、黄芩汤高、低剂量组,每组12只,除正常组外其余小鼠均给予2次腹腔注射10 mg·kg^(-1)氧化偶氮甲烷(AOM),1.5%葡聚糖硫酸钠(DSS)自由饮用7 d,正常饮水2周,“DSS-饮水”重复2个循环。在给予DSS同时,正常组和模型组小鼠给予纯水等剂量灌胃;美沙拉嗪组小鼠给予150 mg·kg^(-1)·d(-1)美沙拉嗪混悬液灌胃;黄芩汤高、低剂量组小鼠分别给予黄芩汤18、9 g·kg^(-1)·d(-1)灌胃;各组均每日给药1次,直至3个循环结束。干预结束后,测量各组小鼠体质量、结肠长度、记录小鼠结肠肿瘤数目,并进行疾病活动指数(DAI)评分。酶联免疫吸附测定法(ELISA)检测血清中白细胞介素(IL)-6、肿瘤坏死因子-α(TNF-α)、IL-1β、IL-4、IL-10及胃肠道肿瘤标志物糖类抗原-199(CA199)含量;苏木素-伊红(HE)染色观察结肠病变;免疫组化法观察结肠增殖细胞相关抗原(Ki67)表达变化;流式法检测小鼠血浆中T淋巴细胞亚群(CD3^(+)、CD4^(+)、CD8^(+)、CD49b^(+))表达;荧光标记法检测小鼠血清中异硫氰酸荧光素-D(FITC-D)含量;蛋白免疫印迹法(Western blot)检测结肠组织细胞周期相关蛋白细胞周期蛋白D1(Cyclin D1)、细胞周期蛋白依赖性激酶-2(CDK2)、细胞周期蛋白依赖性激酶4(CDK4)、紧密连接相关蛋白咬合蛋白(Occludin)、闭合蛋白-1(Claudin-1)蛋白表达。结果:与正常组比较,模型组小鼠体质量下降,DAI评分明显升高,结肠长度变短,IL-6、TNF-α、IL-1β等促炎因子均升高,其中IL-6、TNF-α升高明显(P<0.05)。抑炎因子IL-4明显降低(P<0.05),IL-10降低;肿瘤标志物CA199显著升高(P<0.01);HE染色结果显示模型组小鼠结肠出现瘤性病变,肠黏膜上皮缺损�Objective:To investigate the efficacy of Huangqintang on mouse models of colitisassociated colon cancer(CAC)and explore the mechanism of Huangqintang in regulating immune function and inflammatory response,inhibiting abnormal cell proliferation,and delaying or inhibiting CAC formation in CAC.Method:C57BL/6J mice were randomly divided into a normal group,model group,mesalazine group,and high-and low-dose Huangqintang groups according to body weight,with 12 mice in each group.Except for the normal group,the rest of the mice were given two intraperitoneal injections of 10 mg·kg^(-1) azomethane(AOM)and allowed to drink 1.5%dextran sodium sulfate(DSS)freely for seven days and water normally for two weeks.Then,two cycles of"DSS-drinking water"were repeated.During the administration of DSS,mice in the normal group and model group were given gavage in equal doses of pure water.Mice in the mesalazine group were given 150 mg·kg^(-1)·d^(-1) mesalamine suspension for gavage,and mice in the high-and low-dose Huangqintang groups were given 18 and 9 g·kg^(-1)·d^(-1) Huangqintang for gavage,respectively.Each group was given one dose daily until the end of three cycles.After the intervention,the body weight,colon length,and number of colon tumors in each group were measured,and disease activity index(DAI)scores were performed.The serum contents of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-4(IL-4),interleukin-10(IL-10),and gastrointestinal tumor marker carbohydrate antigen-199(CA199)were detected by enzyme linked immunosorbent assay(ELISA).The colonic lesions were observed by hematoxylin-eosin(HE)staining.The expression of proliferative cell-associated antigen(Ki67)was observed by immunohistochemistry.The expression of T lymphocyte subsets(CD3^(+),CD4^(+),CD8^(+),and CD49b^(+))in mouse plasma was detected by flow cytometry.Fluorescein isothiocyanate-D(FITC-D)content in mouse serum was detected by fluorescent labeling method.The Western blot method was used to detect the expressio

关 键 词：结直肠癌 炎症 通透性 增殖 黄芩汤 

分 类 号：R2-0[医药卫生—中医学] R22R242R285.5R735.35