安肠汤对TNBS致溃疡性结肠炎模型大鼠TLR4/NF-κB信号通路及粪钙卫蛋白表达的影响  

Effects of Anchang Decoction on TLR4/NF-κB Signaling Pathway and the Expression of Fecal Calprotectin in Rats with Ulcerative Colitis Induced by TNBS

作　　者：梁运特 黄仲海[3] 廖志远 张琴 汤勇 孙平良[2] LIANG Yunte;HUANG Zhonghai;LIAO Zhiyuan;ZHANG Qin;TANG Yong;SUN Pingliang(Postgraduate College,Guangxi University of TCM,Nanning 530200,China;Dept.of Gieneral Surgery,the First Affiliated Hospital of Guangxi University of TCM,Nanning 530023,China;Hospital Office,the First Affiliated Hospital of Guangxi University of TCM,Nanning 530023,China)

机构地区：[1]广西中医药大学研究生院,南宁530200 [2]广西中医药大学第一附属医院普外科,南宁530023 [3]广西中医药大学第一附属医院院办,南宁530023

出　　处：《中国药房》2021年第2期189-194,共6页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81660795);广西自然科学基金资助项目(No:2017GXNSFAA198303)。

摘　　要：目的:研究安肠汤对2,4,6-三硝基苯磺酸(TNBS)致溃疡性结肠炎(UC)模型大鼠Toll样受体4(TLR4)/核因子κB(NF-κB)信号通路及粪钙卫蛋白(FC)表达的影响。方法:将SD大鼠随机分为空白组、模型组、阳性对照组[双歧杆菌活菌胶囊,5 m L(含双歧杆菌活菌0.35 g)]和安肠汤低、中、高剂量组(1、5、10 mL,每mL约相当于生药总量0.11 g),每组15只。除空白组大鼠注入生理盐水外,其余各组大鼠均采用TNBS结合乙醇复合灌肠法建立UC模型。造模成功2 d后,空白组、模型组大鼠灌胃生理盐水5mL,各药物组大鼠灌胃相应药物,每日1次,连续给药14 d。末次给药后,采用苏木精-尹红染色法观察大鼠结肠组织的病理改变;采用Western blotting法检测大鼠结肠组织中TLR4、TNF受体关联因子6(TRAF6)、NF-κB蛋白的表达水平;采用实时荧光定量聚合酶链式反应法检测TLR4、TRAF6、TNF-α、NF-κB m RNA的表达水平;采用酶联免疫吸附测定法检测大鼠血清TNF-α、IL-6以及粪便样品中FC的水平。结果:与空白组比较,模型组大鼠的结肠黏膜严重受损,其结肠组织中TLR4、TRAF6、NF-κB蛋白和TLR4、TRAF6、TNF-α、NF-κb m RNA的表达水平以及血清TNF-α、IL-6及粪便样品中FC水平均显著升高(P<0.05);与模型组比较,各药物组大鼠结肠组织病变均有不同程度的改善,且上述指标水平均显著降低(P<0.05)。结论:安肠汤可能通过调节TLR4/NF-κB信号通路及FC水平来减轻UC模型大鼠的炎症反应。OBJECTIVE:To study the effects of Anchang decoction on TLR4/NF-κB signaling pathway and the expression of fecal calprotectin(FC)in TNBS-induced ulcerative colitis(UC)model rats.METHODS:SD rats were randomly divided into blank group,model group,positive control group[Live Bifidobacterium capsules,5 mL(containing Bifidobacterium 0.35 g)],Anchang decoction low-dose,medium-dose and high-dose groups(1,5,10 m L,each milliliter is approximately equivalent to 0.11 g of total crude drug),with 15 rats in each group.Other groups were given TNBS combined with ethanol enema to establish UC model rat,except blank group was given normal saline.Two days after successful modeling,blank group and model group were given normal saline 5 m L,administration groups were given relevant medicine intragastrically,once a day,for consecutive 14 d.After last medication,HE staining was used to observe the pathological change of colon tissue in rats.Western blotting assay was used to detect the protein expression of TLR4,TRAF6 and NF-κB in colon tissues of rats;Real-time fluorescent quantitative PCR was used to detect m RNA expression of TLR4,TRAF6,TNF-αand NF-κB;ELISA assay was adopted to detect serum level of TNF-α,IL-6 and FC in stool samples.RESULTS:Compared with blank group,the colonic mucosa of model group was severely damaged,and the protein expression of TLR4,TRAF6 and NF-κB,m RNA expression of TLR4,TRAF6,TNF-αand NF-κb as well as serum levels of TNF-α,IL-6 and FC level in stool samples were increased significantly(P<0.05).Compared with model group,the pathological changes of colon tissue in rats were improved in different administration groups to different extents,and above indexes were all decreased significantly(P<0.05).CONCLUSIONS:Anchang decoction may relieve the inflammation of UC model rats by regulating the TLR4/NF-κB signaling pathway and the expression of FC.

关 键 词：溃疡性结肠炎 安肠汤 Toll样受体4 TNF受体关联因子6 核因子ΚB 粪钙卫蛋白 

分 类 号：R285.5[医药卫生—中药学]