不同日龄新生SD大鼠坏死性小肠结肠炎模型建立

doi:10.3969/j.issn.1674-5817.2019.06.005

摘要/Abstract

摘要： 目的建立并比较二种不同日龄新生SD大鼠坏死性小肠结肠炎(NEC)模型。方法出生2 h内及出生48 h SD大鼠各24只,随机分为人工喂养组(A、B组)、人工喂养+脂多糖（LPS）组(C、D组）和对照组(E、F组）,即出生2 h的A、C、E组及48 h的B、D、F组。A、B、C、D组均予以缺氧+冷刺激每日2次; C、D组每日LPS灌胃1次; 对照组仅母鼠喂养。实验84 h后空腹处死,取出回盲部近端肠管1 cm,HE染色后观察肠组织形态学,进行病理评分。结果实验组大鼠相继出现不同程度排黄绿色黏液稀便、腹胀、胃潴留、呕吐、嗜睡,活动度下降,并逐渐加重,体质量增长缓慢;对照组新生大鼠纳乳好,生长发育良好。A、B、C、D组组织学病理积分和NEC发生率均明显高于对照组。结论新生SD大鼠出生2 h内及出生48 h在人工灌胃喂养、低体温和缺氧条件下可诱导出符合NEC临床表现和组织学特征的大鼠模型,LPS灌胃可加重临床表现和组织学病理改变。

关键词: 坏死性小肠结肠炎(NEC), 模型, 大鼠, 日龄

Abstract: Objective To establish and compare two models of necrotizing enterocolitis (NEC) in neonatal SD rats at different ages. Methods New born SD rats within postal 2 hours and 48 hours (each time point 24 rats) were randomly divided into three groups respectively according to body weight as the artificial feeding group (groups A, B), the artificial feeding + lipopolysaccharide (LPS) group (groups C, D), and control group (groups E, F), namely postal 2 hours (groups A, C, E) and postal 48 hours (group B, D, F). The rats in 4 groups A, B, C and D were treated with hypoxia plus cold stimulation twice a day, group C and D were also given LPS once a day by gavage, and the control group was fed only by female rats. The rats were executed 84 hours after the experiment time point with fasting. The proximal ileocecal ileum was removed about 1 cm for histomorphological observation. The intestinal histomorphology was observed and the intestinal histopathological score was made. Results The rats in group A, B, C and D had yellow-green mucus defecation, abdominal distension, slow weight growth, gastric retention, vomiting, lethargy, decreased activity and gradually aggravated. Neonatal rats in two control groups had better milk intake, good growth and development. The histological scores and the incidence of NEC in groups A, B, C, and D were significantly higher than those of control groups. Conclusion NEC-compliant models both can be induced in neonatal SD rats within postal 2 hours and 48 hours by artificial nasal feeding, low body temperature and hypoxia, LPS gavage can aggravate the clinical manifestations and histopathological changes.

Key words: Necrotizing enterocolitis(NEC), Models, Rat, Age

中图分类号:

Q95-33

潘兆军, 田兆方, 武荣. 不同日龄新生SD大鼠坏死性小肠结肠炎模型建立[J]. 实验动物与比较医学, 2019, 39(6): 449-453.

PAN Zhao-jun, TIAN Zhao-fang, WU Rong. Establishment of Necrotizing Enterocolitis Model in SD Rats with Different Ages[J]. Laboratory Animal and Comparative Medicine, 2019, 39(6): 449-453.

参考文献

[1] Qian T, Zhang R, Zhu L, et al.Necrotizing enterocolitis in low birth weight infants in China: Mortality risk factors expressed by birth weight categories[J]. Pediatri Neonatol, 2017, 58(6):509-515.
[2] Anderson J G, Rogers EE, Baer RJ, et al.Racial and ethnic disparities in preterm infant mortality and severe morbidity: a population-based study[J]. Neonatology, 2018, 113(1):44-54.
[3] Hull MA, Fisher JG, Gutierrez IM, et al.Mortality and management of surgical necrotizing enterocolitis in very low birth weight neonates: a prospective cohort study[J]. J Am Coll Surg, 2014, 218(6):1148-1155.
[4] Berkhout DJC, Klaassen P, Niemarkt HJ, et al.Risk factors for necrotizing enterocolitis: a prospective multicenter case-control study[J]. Neonatology, 2018, 114(3):277-284.
[5] Klein RM.Models for the study of cell proliferation in the developing gastrointestinal tract[J]. J Pediatr Gastroenterol Nutri, 1986, 5(4):513-516.
[6] Zhou W, Li W, Zheng XH, et al.Glutamine downregulates TLR-2 and TLR-4 expression and protects intestinal tract in preterm neonatal rats with necrotizing enterocolitis[J]. J Pediatr Surg, 2014, 49(7):1057-1063.
[7] 石永言, 富建华, 姚丽, 等. 维生素D调节坏死性小肠结肠炎新生大鼠肠道上皮occludin蛋白表达的研究[J]. 中国小儿急救医学, 2017, 24(1):60-64.
[8] Ares GJ, Mcelroy SJ, Hunter CJ.The science and necessity of using animal models in the study of necrotizing enterocolitis.[J]. Semin Pediatr Surg, 2018, 27(1):29-33.
[9] Cigsar EB, Cetin K, Canan T, et al.The protective effects of sesamol in a neonatal rat model of necrotizing enterocolitis[J]. J Matern Fetal neonatal Med, 2018, 31(1):1-6.
[10] Ling X, Linglong P, Weixia D, et al.Protective effects of bifidobacterium on intestinal barrier function in LPS-induced enterocyte barrier injury of Caco-2 monolayers and in a rat NEC model[J]. PLoS One, 2016, 11(8):e0161635.
[11] Nakame K, Kaji T, Mukai M, et al.The protective and anti-inflammatory effects of glucagon-like peptide-2 in an experimental rat model of necrotizing enterocolitis[J]. Peptides, 2016, 75:1-7.
[12] Do YJ, Sul JW, Jang KH, et al.A novel RIPK1 inhibitor that prevents retinal degeneration in a rat glaucoma model[J]. Ex Cell Res, 2017, 359(1):30-38.
[13] 高晓燕, 李思涛, 肖昕. 坏死性小肠结肠炎诱导新生大鼠全身炎症反应综合征动物模型的建立[J]. 中华实用儿科临床杂志, 2012, 27(11):873-875.
[14] 李美雪, 周伟, 吕回, 等. 脂多糖联合人工喂养诱导新生大鼠坏死性小肠结肠炎模型的建立与评价[J]. 发育医学电子杂志, 2014, 2(1):21-27.
[15] Ni o DF, Sodhi CP, Hackam DJ. Necrotizing enterocolitis: new insights into pathogenesis and mechanisms[J]. Nat Rev Gastroenterol Hepatol, 2016, 13(10):590-600.
[16] Denning TW, Bhatia AM, Kane AF, et al.Pathogenesis of NEC: role of the innate and adaptive immune response[J]. Semin Perinatol, 2017, 41(1):15-28.
[17] 陈晓晴, 许华, 罗小平. 促红细胞生成素对坏死性小肠结肠炎TLR4、NF-κB及IL-6变化的影响[J]. 解放军医学杂志, 2011, 36(9):918-922.
[18] Meng D, Zhu W, Shi HN, et al.Toll-like receptor-4 in human and mouse colonic epithelium is developmentally regulated: a possible role in necrotizing enterocolitis[J]. Pediatr Res, 2015, 77(3):416-424.