Wang Tingting,Xu Lingqi,Gong Yuan,et al.Comparisons of animal models of Hirschsprung-associated enterocolitis[J].Journal of Clinical Pediatric Surgery,,():830-836.[doi:10.3760/cma.j.cn101785-202402017-006]
Comparisons of animal models of Hirschsprung-associated enterocolitis
- Keywords:
- Hirschsprung Disease; Enterocolitis; Disease Models; Animal; Program Evaluation; Comparative Study
- Abstract:
- Objective To employ different modeling approaches for animal models of Hirschsprung-associated 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 1x109 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 obvious 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 expression 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 clinical manifestations. Conclusions The chemically induced model of E.coli JM83 gavage after benzalkonium chloride wrapping and spontaneous animal model after EdnrB gene knockout both exhibit clinical and pathological characteristics consistent with HAEC,making them suitable for elucidating the etiology and pathogenesis of HAEC.However,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.
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Memo
收稿日期:2024-2-20。
基金项目:苏州市临床病种诊疗技术专项(LCZX202107);苏州市重点学科(SZXK202105)
通讯作者:黄顺根,Email:drhuang01@163.com