三硝基苯磺酸诱导小鼠溃疡性结肠炎模型制备的技术改良

Preparation of a mouse model of TNBS-induced ulcerative colitis:technology improvement and optimal dose exploration

目的:改良2,4,6-三硝基苯磺酸(TNBS)硅胶管灌肠诱导制备小鼠溃疡性结肠炎(UC)模型的方法,提高造模的成功率和模型的稳定性,并探索造模的适宜剂量和时间.方法:选用40只SPF级♂Balb/c小鼠,6-8周龄,随机分为正常对照组、不同浓度TNBS组(37.5mg/kg、75mg/kg、150mg/kg、200mg/k g),每组8只,使用"灌胃针"替代"硅胶管"灌肠,并于灌肠后2d和4d分别处死4只小鼠,观察不同组别小鼠生理状态、结肠组织的损伤及病理学的改变情况.结果:在"灌胃针"造模过程中未发生小鼠死亡现象;对照组小鼠一般情况及结肠黏膜组织无异常改变;小鼠灌肠后出现少食、少动、体质量下降、皮毛光泽度下降、腹泻、便血.不同浓度TNBS造模组随着TNBS剂量的增加,小鼠结肠黏膜组织出现充血、出血、水肿、炎症、溃疡的程度增加.HE染色可见结肠组织水肿、炎症细胞浸润、杯状细胞缺失、溃疡形成的程度逐渐增加.其中TNBS37.5mg/kg、75mg/kg组于造模后2d,以上损伤现象开始缓解,未形成稳定的UC模型;150mg/kg、200mg/kg组持续时间较长,以上损伤现象4d内未见明显缓解,150mg/kg组表现为较典型的UC模型,200mg/kg为重症UC模型.结论:对制造小鼠UC模型进行相关技术改进,使灌肠更加简便,提高造模效率,显著增加了模型的稳定性.

AIM：To establish a mouse model of 2,4,6-trinitrobenzenesulfonic acid （TNBS）-induced colitis using the ＂gavage needle＂ instead of ＂silicone tube＂,and to explore the optimal dosage and induction time of TNBS.METHODS：Forty SPF male Balb/c mice were randomly and equally divided into five groups：normal control group and four TNBS groups （treated with 37.5,75,150,and 200mg/kg of TNBS）.Four mice in each group were killed on days 2 and 4 after model induction to observe the physical status and evaluate pathologic changes in the colon.RESULTS：No death occurred during the ＂gavage needle＂-based modeling preparation.The control group showed no abnormalities.Higher doses of TNBS induced more severe congestion,hemorrhage,edema,inflammation and ulcer in the colon mucosa.Colonic tissue edema,inflammatory cell infiltration,goblet cell deletion,and formation of ulcer were aggravated significantly as the dosage of TNBS increased.The above mentioned features recovered from day 2 after model induction in mice treated with 37.5 and 75mg/kg of TNBS,but showed no obvious alterations on day 4 after model induction in mice treated with 150 and 200mg/kg of TNBS.CONCLUSION：The ＂gavage needle＂-based method could improve efficiency and increase stability in preparing the mouse model of TNBSinduced ulcerative colitis.