2,4,6-三硝基苯磺酸与不同浓度乙醇诱导大鼠溃疡性结肠炎模型的建立及其稳定性评价

Establishment and stability of ulcerative colitis rat model induced by 2,4,6-trinitrobenzene sulfonic acid

目的建立2,4,6-三硝基苯磺酸(TNBS)-乙醇混合液诱导大鼠溃疡性结肠炎模型并判断不同浓度的乙醇对模型稳定性的影响。方法将80只雄性SD大鼠,随机分为空白组、模型Ⅰ组、模型Ⅱ组、模型Ⅲ组共4组,每组各20只,模型Ⅰ、Ⅱ、Ⅲ组大鼠分别给予TNBS(100 mg·kg^(-1))-无水乙醇0.25 mL、TNBS(100 mg·kg^(-1))-50%乙醇0.25 mL、TNBS(100 mg·kg^(-1))-25%乙醇0.25 mL混合液灌肠,空白组大鼠给予等量生理盐水灌肠。造模后每日观察大鼠精神状态及死亡情况,计算大鼠的疾病活动指数(DAI),并于2、7、14、21、28 d分批处死相同数量的大鼠,采集结肠组织,计算结肠黏膜损伤指数(CMDI),同时进行HE染色,计算组织学损伤指数(TDI)。结果除空白组外,各模型组造模后均有明显的溃疡性结肠炎临床症状和病理学改变(P<0.05);各模型组的死亡率、DAI、CMDI、TDI评分均在造模后7 d高于其他时间点,且差异有统计学意义(P<0.05);模型Ⅲ组无死亡,与模型Ⅰ组、模型Ⅱ组相比,DAI、CMDI、TDI评分差异有统计学意义(P<0.05)。结论模型Ⅰ、Ⅱ、Ⅲ组均能成功诱导溃疡性结肠炎模型,以模型Ⅲ组死亡率较低、模型稳定性较好。

Objective To establish the ulcerative colitis （UC） model in rats induced by 2, 4, 6-trinitrobenzene sulfonic acid （TNBS） and determine the effect of ethanol concentration on the stability of the model rat. Methods Totally 80 male SD rats were randomly divided into 4 groups （a blank group, and model Ⅰ, Ⅱ, Ⅲ group, 20 rats in each group）. Rats in the model group I, the model group Ⅱ, the model group Ⅱ were respectively given TNBS （100 mgokg - 1）- 0.25 mL anhydrous ethanol, TNBS （100 mg·kg - 1）-0.25 mL 50% ethanol and TNBS （100 mg kg - 1）-0.25 mL 25% ethanol mixture enema. The blank group was given an equal amount of saline enema. After modeling, the mental status and mortality of rats were observed every day, and the disease activity index （DAI） of rats was calculated. After the replicating the ulcerative colitis model successfully, the rats were sacrificed on day 2, 7, 14, 21 and 28, so that the colonic tissue was collected and the colon mucosa damage index （CMDI） was calculated. Hematoxylin eosin staining was performed to calculate the histopathological damage index （TDI）. Results The clinical symptoms and pathological changes of UC were observed in all the model groups （P 〈 0.05）. The mortality, DAI, CMDI and TDI scores of the model group were all higher on day 7 than those at other time points （P 〈 0.05）. The model group Ⅲhad no death, the DAI, CMDI, and TDI scores were different from those of the model group Ⅰ and Ⅱ （P 〈 0.05）. Conclusion Ulcerative colitis models are successfully induced, and the model group Ⅲ as the most appropriate, with lower mortality rate and good stability.