临床小儿外科杂志2024，Vol.23Issue(9)：830-836,7.DOI:10.3760/cma.j.cn101785-202402017-006
先天性巨结肠相关性小肠结肠炎动物模型造模方法的对比研究
Comparisons of animal models of Hirschsprung-associated enterocolitis
摘要
Abstract
Objective To employ different modeling approaches for animal models of Hirschsprung-associ-ated enterocolitis(HAEC)to identify a more dependable model for elucidating the pathogenesis of HAEC.Methods A total of 24 21-day-old Sprague Dawley(SD)rats were randomized into experimental group treated with 0.5％benzalkonium chloride and control group treated with saline.After 3-week modeling,experimental group was gavaged with E.coli JM83 1×10 9 CFU/d,and control group an equivalent amount of saline.Two groups were subsequently compared in terms of diet,activity and defecation.Additionally,21-day-old EdnrB-/-mice were selected as experimental group and wild-type littermates as control group(n=5 each).Intestinal segments of mice in experimental group were identified as narrowed and dilated segments according to gross morphology.Pathological changes were evaluated by hematoxylin-eosin(HE)stain and altered expressions of intestinal-related barrier proteins by Western blot.Results HAEC model was successfully established by gavage with E.coli after a treatment of benzalkonium chloride.And 21-day-old EdnrB-/-mice could also be utilized as an animal model of HAEC.As compared with control groups,experimental groups of both animal models showed slow activity and ob-vious abdominal expansion.There were obvious narrowing of distal intestine and dilatation of proximal intestine.HE stain revealed a significant absence of ganglion cell clusters in narrow segment in both animal models along with a noticeable aggregation of inflammatory cells in dilated segment.No obvious abnormality was detected in control group.Western blot analysis revealed that in HAEC patients,the relative expression levels of E-Cadherin protein in dilated segment were significantly lower than those in control group[(0.15±0.01)vs.(1.13±0.08),t=12.940,P＜0.001].Similarly,Occludin protein expression was also lower in HAEC dilated segment than that in control group[(0.21±0.01)vs.(0.99±0.01),t=95.030,P＜0.001].In benzalkonium chloride-induced model,Occludin protein relative expression was lower in dilated segment than that in control group[(0.14±0.01)vs.(0.94±0.04),t=12.020,P＜0.001]while Claudin3 protein expression was lower than control group[(0.34±0.01)vs.(0.99±0.01),t=38.240,P＜0.001].In EdnrB-/-murine model,E-Cadherin protein ex-pression was lower in dilated segment than that in control group[(0.28±0.01)vs.(0.97±0.03),t=25.360,P＜0.001]and Occludin protein expression was lower than control group[(0.32±0.01)vs.(1.13±0.02),t=43.710,P＜0.001].Additionally,Claudin3 protein expression was,lower than control group[(0.17±0.01)vs.(1.19±0.03),t=36.960,P＜0.001].These differences were statistically significant and consistent with clini-cal manifestations.Conclusions The chemically induced model of E.coli JM83 gavage after benzalkonium chlo-ride wrapping and spontaneous animal model after EdnrB gene knockout both exhibit clinical and pathological char-acteristics consistent with HAEC,making them suitable for elucidating the etiology and pathogenesis of HAEC.How-ever,EdnrB-/-mice,as gene knockout mice relevant to the pathogenesis of HD,may better mimic the clinical course development and physiopathological changes of HAEC by not intervening in natural occurrence of enterocolitis.关键词
Hirschsprung病/小肠结肠炎/疾病模型,动物/方案评价/对比研究Key words
Hirschsprung Disease/Enterocolitis/Disease Models,Animal/Program Evaluation/Comparative Study引用本文复制引用
王婷婷,徐铃琪,龚元,李娜,杨献,周慧婷,黄顺根..先天性巨结肠相关性小肠结肠炎动物模型造模方法的对比研究[J].临床小儿外科杂志,2024,23(9):830-836,7.基金项目
苏州市临床病种诊疗技术专项(LCZX202107) （LCZX202107）
苏州市重点学科(SZXK202105) Key Disease Diagnosis and Treatment Project of Suzhou Municipal Health & Family Plan-ning Commission(LCZX202107) （SZXK202105）
Suzhou Municipal Key Discipline of Medicine(SZXK202105) （SZXK202105）
