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大鼠实验性坏死性小肠结肠炎实验方案的验证及操作过程中的陷阱

Validation of protocol of experimental necrotizing enterocolitis in rats and the pitfalls during the procedure.

作者信息

Gonçalves Frances Lilian Lanhellas, Gallindo Rodrigo Melo, Soares Lucas Manoel Mangueira, Figueira Rebeca Lopes, Volpe Fábio Antônio Perecim, Pereira-da-Silva Maurício André, Sbragia Lourenço

机构信息

Division of Pediatric Surgery, Department of Surgery and Anatomy, School of Medicine of Ribeirao Preto, University of Sao Paulo, Ribeirao Preto, SP, Brazil.

出版信息

Acta Cir Bras. 2013;28 Suppl 1:19-25. doi: 10.1590/s0102-86502013001300005.

Abstract

PURPOSE

To describe the difficulties of implementing the protocol of experimental necrotizing enterocolitis (NEC) in order to obtain a larger number of newborns affected with the disease and a lower mortality.

METHODS

Term Sprague-Dawley newborns rats (22 days) were divided into four groups of 12 fetuses each (n = 48): EC--breastfed newborns; IH--breastfed newborns and subjected to a stress protocol by ischemia and hypothermia; ESB--formula-fed newborns (Esbilac®, PetAg, Hampshire, IL, USA) and NEC--formula-fed newborns and subjected to stress protocol. The parameters set for the study protocol were: milk concentration (0.19 g ml or 0.34 g/ml), diet instilled volume (according to body weight--200 kcal/day/Kg--or progressive, according to acceptance), weight (gain, loss or maintenance) and duration of the experiment (72 hours or 96 hours). Data of body weight (BW), intestinal weight (IW) and the IW/BW ratio were obtained. Samples of terminal ileum were collected and analyzed by the degree of injury to the intestinal wall. Statistically significance was set to p<0.05.

RESULTS

The established protocol with less mortality and increased number of NEC was with Esbilac® at a concentration of 0.19 g/ml of diet instilled volume of 0.1 ml, every 3 hours, for 72 hours. All infants fed with artificial milk lost weight. In the degree score of intestinal injury, the ESB, IH and NEC groups were considered positive for NEC with greater histological injury in the latter.

CONCLUSION

The described NEC protocol in rats allowed a greater survival of puppies with a greater number of animals affected by the disease.

摘要

目的

描述实施实验性坏死性小肠结肠炎(NEC)方案的困难,以获得更多患该病的新生儿并降低死亡率。

方法

足月的斯普拉格-道利新生大鼠(22日龄)被分为四组,每组12只胎儿(n = 48):EC组——母乳喂养的新生儿;IH组——母乳喂养的新生儿并接受缺血和低温应激方案;ESB组——人工喂养的新生儿(使用美国伊利诺伊州汉普郡PetAg公司的Esbilac®奶粉);NEC组——人工喂养的新生儿并接受应激方案。研究方案设定的参数包括:奶浓度(0.19 g/ml或0.34 g/ml)、喂食量(根据体重——200 kcal/天/千克——或根据接受情况逐步增加)、体重(增加、减轻或维持)以及实验持续时间(72小时或96小时)。获取体重(BW)、肠重(IW)和IW/BW比值的数据。收集回肠末端样本并根据肠壁损伤程度进行分析。统计学显著性设定为p<0.05。

结果

死亡率较低且NEC数量增加的既定方案是使用浓度为0.19 g/ml的Esbilac®奶粉,每3小时喂食0.1 ml,持续72小时。所有人工喂养的婴儿体重均减轻。在肠损伤程度评分中,ESB组、IH组和NEC组被认为患有NEC呈阳性,其中NEC组的组织学损伤更严重。

结论

所描述的大鼠NEC方案使患病动物数量更多的幼犬存活率更高。

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