三硝基苯磺酸诱导急性肠纤维化大鼠模型的改良方法  

Modified Method for Inducing Acute Intestinal Fibrosis in Rats Using 2,4,6-Trinitrobenzene Sulfonic Acid

作　　者：王怡如 蒋笑影 董若曦 潘一滨[1] 韩向晖 曹永清[1] WANG Yiru;JIANG Xiaoying;DONG Ruoxi;PAN Yibin;HAN Xianghui;CAO Yongqing(Department of Anal-rectal Surgery,Longhua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;Department of Institute of Chinese Traditional Surgery,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属龙华医院肛肠科,上海200032 [2]上海中医药大学附属龙华医院中医外科研究所,上海200032

出　　处：《实验动物与比较医学》2022年第4期284-293,共10页Laboratory Animal and Comparative Medicine

基　　金：国家自然科学基金资助项目(81874469)“红萸饮下调chr10:115386962-115390436+/miR-6914-5p/Atg7通路治疗IBD的机制研究”。

摘　　要：目的探索2,4,6-三硝基苯磺酸(2,4,6-trinitrobenzene sulfonic acid,TNBS)诱导急性肠纤维化大鼠模型的方法及不同药物配比的效果。方法30只雌性SD大鼠随机分为5组,每组各6只。A~D组为模型组,模型组大鼠采用不同配比的药物溶液通过改良保留灌肠法一次性造模:A组,5%TNBS+50%乙醇(体积比1∶1);B组,5%TNBS+75%乙醇(体积比1∶1);C组,5%TNBS+100%乙醇(体积比1∶1);D组,5%TNBS+50%乙醇(体积比2∶1);对照组用生理盐水(0.9%氯化钠溶液)灌肠;观察造模后1周内动物的症状、体征和体质量变化,并进行疾病活动指数(disease activity index,DAI)评分。分别于造模后7 d和14 d,随机选择每组内一半大鼠进行取材,观察结肠组织大体损伤程度并以结肠炎损伤指数(colon macroscopic damage index,CMDI)评分;结肠组织病理切片以HE染色观察肠炎严重程度,Masson染色观察胶原纤维沉积情况。结果各模型组大鼠均出现轻重不等的肠炎及肠纤维化病变,其中以体积比为1∶1的5%TNBS+75%乙醇溶液造模观察期内未导致大鼠死亡,造模后24 h大鼠体质量明显下降、出现稀便及血便症状,结肠壁纤维化病变明显,体质量下降、DAI评分与CDMI评分升高,与对照组相比差异具有统计学意义(P<0.05);光学显微镜下炎症程度较重,呈透壁性;Masson染色显示造模部位的肠壁增厚明显,黏膜下层、肌层及浆膜层均出现弥漫性胶原纤维沉积。结论改良的TNBS保留灌肠法能有效构建肠纤维化大鼠模型,体积比为1∶1的5%TNBS+75%乙醇溶液所建模型可模拟克罗恩病纤维化两大特征,即透壁性炎症和肠壁纤维化。该造模方法简便、高效,动物死亡率低。Objective To explore 2,4,6-trinitrobenzene sulfonic acid(TNBS)-induced acute intestinal fibrosis in rats and the effect of different drug ratios.Methods Thirty female Sprague-Dawley(SD)rats were randomly divided into five groups with six rats in each group.Groups A to D were the model groups,in which the rats were induced by modified retention enemas using solutions with different drug ratios:group A,5%TNBS+50%ethanol(1∶1 v/v);group B,5%TNBS+75%ethanol(1∶1 v/v);group C,5%TNBS+100%ethanol(1∶1 v/v);group D,5%TNBS+50%ethanol(2∶1 v/v);and the control group was induced by normal saline(0.9%sodium chloride solution)enema.The symptoms,signs,and body mass changes of the animals were observed within one week after modeling,and scored using the disease activity index(DAI).At 7th d and 14th d after model establishment,half of the rats in each group were randomly selected for sampling to observe the degree of gross damage to the colon tissue and scored using the colon macroscopic damage index(CMDI).Pathological sections of colon tissue were stained with hematoxylin-eosin(HE)to observe the severity of enteritis,and Masson staining was used to observe collagen fiber deposition.Results The rats in each model group showed enteritis and intestinal fibrosis lesions of different severities,of which 5%TNBS+75%ethanol solution(1∶1 v/v)did not lead to death during the observation period.At 24 h after model establishment,the rats had significantly decreased body weight,loose stool,and bloody stool,significant colonic wall fibrosis lesions,and increased DAI and CDMI scores compared with the control group(P<0.05).The degree of inflammation was transmural and more severe,as seen under the light microscope,and Masson staining showed that the intestinal wall at the model site was significantly thickened,and diffuse collagen fiber deposition occurred in the submucosa,muscular layer,and serosal layer.Conclusion The modified TNBS retention enema method can effectively construct a rat model of intestinal fibrosis,and the model establi

关 键 词：炎症性肠病 克罗恩病 肠道纤维化 2 4 6-三硝基苯磺酸 SD大鼠 

分 类 号：R-332[医药卫生] Q95-33[生物学—动物学]