坏死性小肠结肠炎晚发和长病程动物模型的建立  

A late-onset and long-term model of necrotizing enterocolitis in rat pups

作　　者：王玲玲[1] 金芳[1] 温博贤[1] 冼其 熊慧[1] 林楚琴[1] 黄堃莹 江观银 黄艳[1] 

机构地区：[1]中山大学附属第六医院儿科,广东广州510655 [2]广州市红十字会医院病理科,广东广州510220

出　　处：《当代医学》2018年第8期1-5,共5页Contemporary Medicine

基　　金：广东省医学科研基金(A2013230);广东省科技计划项目(2013B021800119)

摘　　要：目的建立坏死性小肠结肠炎(NEC)晚发和长病程动物模型,为NEC相关研究提供新的实验平台。方法 10日龄SD大鼠随机分为Dith/FFLAS组(n=47)、FFLAS组(n=32)、Dith组(n=32)、Control组(n=32)。Dith/FFLAS组给予腹腔注射双硫腙(Dith),结合人工喂养和缺氧、内毒素LPS刺激(FFLAS)进行诱导。每天观察临床表现和体质量变化,于24 h、72 h、7 d、15 d抽样进行肠组织学评分,分析各时点的NEC患病率和病损程度,评估最佳成模时间和病变迁延情况。结果 Dith/FFLAS组48~72 h普遍出现腹胀、腹泻等NEC症状,个别出现血便;72 h和7 d患病率最高,分别为87.50%(7/8)和71.43%(5/7),显著高于FFLAS组(P<0.05)。4组肠组织评分分别为:24 h(1.00±0.53),(0.50±0.53),(0.50±0.53),(0.25±0.46),72 h(2.38±0.92),(1.13±0.64),(0.25±0.46),(0.38±0.52),7 d(2.00±0.82),(0.75±0.46),(0.13±0.35),(0.25±0.46),15 d(1.00±0.63),(0.38±0.52),(0.25±0.46),(0.13±0.35),差异均有统计学意义(F24 h=3.00,F72 h=17.05,F7 d=14.09,F15 d=4.08,P值均<0.05)。各时点均以Dith/FFLAS组评分最高,不仅与Control组差异有统计学意义(P<0.05);而且在72 h、7 d、15 d时点都显著高于FFLAS组(P<0.05)和Dith组(P<0.05)。Dith/FFLAS组不同时点病变程度差异有统计学意义(F=6.60,P<0.01);72 h和7 d损伤程度显著高于24 h和15 d(P<0.05)。结论 Dith/FFLAS法能在10日龄大鼠成功建立晚发和长病程NEC模型,72 h成模率87.50%,病变至少迁延至第7天,15天仍未完全修复。Objective To develop a late-onset and long-term NEC model in rat pups. Methods 10-day-old rat pups were randomly divided into four groups: Dith/FFLAS(n=47), FFLAS(n=32), Dith(n=32), and Control(n=32). For group Dith/FFLPS, pups were i.p injected dithizone, followed by FFLAS for 7 days. Clinical symptoms and body weight were recorded daily. At the time points of 24 h, 72 h, 7 d,15 d, ilea were harvested from sampled pups, and histological damage were scored for evaluating NEC morbidity and severity. Results Pups treated with Dith/FFLAS showed abdominal distension and diarrhea, and a few pups defecated bloody stools in 48-72 h. NEC morbidity reach the highest level at points 72 h and 7 d, significantly higher than FFLAS group[e.g 72 h(7/8) vs(2/8), P=0.019; 7 d(5/7) vs(1/8), P=0.034 ]. The histological scores of four groups were: at time point. 24 h[(1.00±0.53),(0.50±0.53),(0.50±0.53),(0.25±0.46)]; 72 h[(2.38±0.92),(1.13±0.64),(0.25±0.46),(0.38±0.52)]; 7 d[(2.00±0.82),(0.75±0.46),(0.13±0.35),(0.25±0.46)]; 15 d[(1.00±0.63),(0.38±0.52),(0.25±0.46),(0.13±0.35)], and the difference among four groups were significant(F24 h=3.00, F72 h=17.05, F7 d=14.09, F15 d=4.08; of each point, P<0.05). At each point, the intestinal damage in group Dith/FFLAS was the most serious of four groups, not only always significantly different from group control(P<0.05), but also obviously severe compairing with group FFLAS and group Dith at point 72 h, 7 d and 15 d respectively(P<0.05). The scores of group Dith/FFLAS at four points were different significantly(F=6.60, P<0.01), higher at point 72 h and 7 d, than that at point 24 h(P<0.05), and 15 d(P<0.05) respectively. Conclusion FFLAS is not a suitable way to induce NEC in 10-day-old rat pups. Whereas, Dith/FFLAS method can develop a late-onset and long-term model of NEC, and the incidence at 72 h were 87.50%, and the lesion last at least to 7 th day, and can't recovery completely at 15 th day.

关 键 词：坏死性小肠结肠炎 人类疾病动物模型 双硫腙 

分 类 号：R-332[医药卫生] R574