一种新型豚鼠反流性食管炎模型的构建  

A novel method for establishing a guinea pig model of reflux esophagitis

作　　者：次仁玉珍 吴静 查兰兰 余晓云[2] 沈磊[1] Ciren Yuzhen;Wu Jing;Zha Lanlan;Yu Xiaoyun;Shen Lei(Department of Gastroenterology,Renmin Hospital of Wuhan University,Wuhan 430060,China;Department of Gastroenterology,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China)

机构地区：[1]武汉大学人民医院消化内科,武汉430060 [2]华中科技大学同济医学院附属协和医院消化内科,武汉430022

出　　处：《中华实验外科杂志》2024年第3期622-626,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81570490,82070555)。

摘　　要：目的探讨不同天数饮用含胃蛋白酶的盐酸(PH=3)的豚鼠建立反流性食管炎模型的效果,并确定最佳天数的新型建模方法。方法将40只3~4周龄的SD雄性豚鼠采用随机数字表法分为对照组、14 d模型组、21 d模型组、28 d模型组、35 d模型组,每组各8只。适应性饲养7 d后开始造模,其中对照组予以饮用无菌水,500 ml/d,连续28 d。模型组予以饮用含0.5%胃蛋白酶的0.1 mol/L盐酸(HCl),PH=3500ml/d,分别连续14、21、28、35 d。每天观察各组豚鼠的一般状况、体重变化。取材后记录食管组织肉眼状态,行苏木精-伊红(HE)染色组织病理学观察。各组豚鼠食管组织通过实时荧光定量聚合酶链反应(RT-PCR)检测白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、肿瘤坏死因子-α(TNF-α)、闭锁小带蛋白-1(ZO-1)和闭合蛋白(Occludin)的mRNA相对表达量。同时蛋白质印迹法(Western blot)检测ZO-1、Occludin的蛋白表达。两组间比较采用t检验。结果相较于对照组,各模型组的豚鼠活动力下降、体重增长缓慢,部分出现腹泻及皮疹。大体形态观察可见各模型组豚鼠的食管组织不同程度的黏膜充血,未见明显糜烂灶。光镜下可见各模型组食管黏膜出现不同程度早期炎症损伤,28 d模型组和35 d模型组可见明显的食管黏膜增厚、炎性细胞浸润、鳞状上皮细胞过度增生、固有层乳头延长等食管炎的表现。随着造模时间的延长,模型组豚鼠食管组织中炎性因子表达增加、黏膜屏障功能受损。从Western blot结果可见,21d、28、35 d模型组的连接蛋白ZO-1的蛋白表达水平均低于对照组(0.612±0.051比1.000±0.157、0.508±0.068比1.000±0.157、0.514±0.094比1.000±0.157,t=4.076、4.979、4.597,P<0.05)。14、21、28、35 d模型组的连接蛋白Occludin的蛋白表达水平均低于对照组(0.758±0.120比1.000±0.089、0.725±0.093比1.000±0.089、0.444±0.054比1.000±0.089、0.443±0.099比1.000±0.089,t=2.802、3.699Objective To explore the effectiveness of establishing a Reflux Esophagitis(RE)model in guinea pigs by ingesting pepsin-containing hydrochloric acid(pH=3)for different durations,determining an optimal duration for a novel modeling approach.MethodsA total of 40 male SD guinea pigs aged 3-4 weeks(purchased from Hubei Yizhicheng Biotechnology Co.,Ltd.)were randomly divided into control,14-day model,21-day model,28-day model,and 35-day model groups,with 8 guinea pigs in each group.After 7 days of adaptation,modeling was initiated.The control group received sterile water(500 ml/day)for 28 consecutive days.The model groups were administered 0.1 mol/L hydrochloric acid containing 0.5%pepsin,with a pH of 3,at 500 ml/day,respectively for 14,21,28,and 35 days,respectively.General conditions and weight changes of the guinea pigs in each group were observed daily.After sample collection,the macroscopic appearance of the esophageal tissue was recorded,and histopathological observations were conducted using hematoxylin-eosin(HE)staining.Quantitative real-time polymerase chain reaction(RT-PCR)was employed to assess the relative mRNA expression levels of interleukin-6(IL-6),interleukin-10(IL-10),tumor necrosis factor-α(TNF-α),Zonulaoccludens-1(ZO-1),and occludin in the esophageal tissues of the guinea pigs.Additionally,protein expression of ZO-1 and occludin was evaluated using Western blotting.Comparison between the two groups was performed by t test.ResultsCompared to the control group,the activity of guinea pigs in each model group decreased,and their weight gain was slow,with some experiencing diarrhea and rash.Macroscopic observations revealed varying degrees of congestion in the esophageal tissues of guinea pigs in each model group,with no apparent ulceration.Under the microscope,early inflammatory damage to the esophageal mucosa was observed in all model groups.The 28-day and 35-day model groups exhibited noticeable thickening of the esophageal mucosa,inflammatory cell infiltration,excessive proliferation of squamous ep

关 键 词：反流性食管炎 自发性饮酸 豚鼠 动物模型 

分 类 号：R571[医药卫生—消化系统] R-332[医药卫生—内科学]