黄连通过调控肠黏膜上皮细胞AQPs蛋白表达改善溃疡性结肠炎腹泻的机制研究  

Mechanism of Rhizoma coptidis in improving diarrhea in ulcerative colitis through regulating the expression of AQPs protein in colonic mucosal epithelial cells

作　　者：王师英 赵文樱子 钟彩铃 马燕颜 陈立基 张北平 WANG Shi-ying;ZHAO Wen-ying-zi;ZHONG Cai-ling;MA Yan-yan;CHEN Li-ji;ZHANG Bei-ping(Department of Gastroenterology,the Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510120,China)

机构地区：[1]广州中医药大学第二附属医院脾胃病科,广州510120

出　　处：《北京中医药》2022年第9期954-958,共5页Beijing Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82174126,81904106);广东省中医院中医药科学技术研究专项课题(YN2020QN15)。

摘　　要：目的基于肠黏膜上皮细胞水通道蛋白(AQPs)表达的调控作用,探究黄连改善溃疡性结肠炎(UC)腹泻的药理机制。方法将SD大鼠随机分为对照、模型、黄连低剂量、黄连中剂量、黄连高剂量组5组,各6只。采用葡聚糖硫酸钠盐(DSS)建立UC大鼠模型,造模结束后第2天,各给药组开始灌胃相应药物,1次/d,连续7 d,计算并评价各组大鼠稀便率、平均稀便级和腹泻指数;采用苏木素伊红(HE)染色法观察各组结肠组织病理变化;应用FITC-Dextran检测结肠黏膜通透性;采用ELISA试剂盒检测各组血清中白细胞介素(IL)1β和肿瘤坏死因子(TNF)-α水平;采用荧光定量PCR法和Western blotting法检测各组结肠黏膜上皮细胞中AQP3和AQP4基因和蛋白表达情况。结果模型组大鼠稀便率、平均稀便级和腹泻指数增加(P<0.01),结肠黏膜通透性及炎症损伤加重;肠黏膜上皮细胞中AQP3和AQP4基因和蛋白表达水平均明显下降(P<0.01)。与模型组比较,黄连中、高剂量组大鼠的稀便率、平均稀便级和腹泻指数降低(P<0.01),且结肠黏膜通透性及炎症损伤改善;肠黏膜上皮细胞中AQP3和AQP4基因和蛋白表达水平升高(P<0.01);其中,黄连中剂量组改善UC大鼠的肠黏膜炎症损伤和腹泻疗效优于黄连高剂量组。结论黄连具有较好的止泻作用,其作用机制可能与减轻UC结肠黏膜炎性损伤,上调肠黏膜上皮细胞中AQP3和AQP4基因和蛋白的表达有关。Objective To explore the pharmacological mechanism of Rhizoma coptidis(Rc)in improving diarrhea in ulcerative colitis based on the regulatory effect of aquaporins(AQPs)in colonic mucosal epithelial cells(cmEC).Methods SD rats were randomly divided into 5 groups,including control group,model group,low,medium and high dose group of Rhizoma coptidis,with 6 rats in each group.Dextran sulfate sodium salt(DSS)was used to establish UC rat model.On the second day after modeling,each group was given by garage the corresponding drugs once a day,while the control and model groups were given the same dose of distilled water.The loose stool rate,the average ranking of loose stool,and the diarrhea index were compared according to the fecal traits and stool times after 7 days of treatment.Hematoxylin-eosin(HE)staining was used to observe the pathological morphological changes of colon tissue;FITC-Dextran was used to detect the colonic mucosal permeability;ELISA kit was used to detect the serum interleukin-1β(interleukin-1β)and tumor necrosis factor-ɑ(TNFɑ)of the rats;the fluorescence quantitative PCR and Western blotting were used to detect the expressions of AQP3 and AQP4 in cmEC of rats.Results The loose stool rate,the average ranking of loose stool,and diarrhea index were significantly increased in the model group(P<0.01),and the colonic mucosal permeability and colonic mucosal inflammatory injury were also considerably increased;meanwhile,the gene and protein expression of AQP3 and AQP4 in cmEC were significantly increased(P<0.01).Compared with the model group,the loose stool rate,average ranking of loose stool,and diarrhea index of the rats in the medium and high dose groups of Rc were significantly ameliorated(P<0.01)with the colonic mucosal permeability and colonic mucosal inflammatory injury improved.At the same time,the gene and protein expression of AQP3 and AQP4 in cmEC were significantly increased(P<0.01).Moreover,the effect of Rc was more significant in the middle-dose group than that in the high dose group of

关 键 词：溃疡性结肠炎 黄连 腹泻 水通道蛋白 大鼠 

分 类 号：R285[医药卫生—中药学]