巴马小型猪十二指肠及回肠末端双瘘管模型构建及评价

doi:10.16431/j.cnki.1671-7236.2025.05.013

摘要/Abstract

摘要： 【目的】为解决现有肠道单瘘动物模型存在体重过大、瘘管易脱落和无法分别探究消化吸收时胃部及肠部作用等问题，试验旨在建立巴马小型猪十二指肠及回肠末端双瘘管模型，探究动物肠道双瘘管模型在营养物质或药物等胃肠道消化吸收相关试验中的可行性和有效性。【方法】选用11只9月龄巴马小型猪(17.5 kg±2.0 kg)，麻醉后在右腹部合适位置切口，暴露肠道并进行荷包缝合，插入瘘管后固定双瘘管并缝合切口。术后严格护理并观察动物情况，防止感染。试验动物术后恢复后，以直接超高温(dUHT)牛乳为研究对象，利用SDS-PAGE、激光共聚焦显微镜(CLSM)、超高效液相色谱(UPLC)等检测肠道流出物和血液中蛋白质及氨基酸变化情况，分析牛乳蛋白在消化吸收过程中的动态变化，确认所建立模型在模拟食物消化吸收中的可靠性和实用性。【结果】11例试验动物在术后2个月内全部存活，无明显不适或并发症，且生长状况良好，动物平均体重增加12.23 kg，符合正常生长趋势。试验期间未观察到瘘管脱落情况，且瘘管功能正常，能够顺利收集十二指肠和回肠的流出物以及血液样本。测试试验中，检测流出物样品蛋白质微观结构，可观察到随着消化时间不断延长，蛋白质含量逐渐减少；消化过程中，酪蛋白和β-乳球蛋白条带呈现先增强后减弱的趋势；检测摄食前后不同时间点血液中氨基酸含量，得到摄食前后氨基酸吸收曲线，摄食后血浆中总氨基酸的曲线下增量面积(iAUC)为(12 105.00±2 145.57) μg·min/mL，符合正常的牛乳蛋白机体吸收动力学。【结论】试验成功建立了巴马小型猪十二指肠及回肠末端双瘘管模型，验证了手术技术和术后护理方案的有效性，证明该双瘘管模型在巴马小型猪中具有良好的应用前景，为胃肠道消化吸收功能的研究提供了可靠的试验动物模型，为探究营养物质的消化吸收、新药研发和评估现有药物的吸收效果等提供了有力支持。

关键词: 巴马小型猪; 双瘘管; 消化吸收; 术后护理

Abstract: 【Objective】 To address the issues of excessive body weight,easy detachment of cannulas and the inability to separately investigate the roles of gastric and intestinal parts in digestion and absorption in the existing single-cannulas animal models,the double cannulas model of duodenum and terminal ileum in Bama Minipigs was established to investigate the feasibility and effectiveness of double-cannulas models of the animal intestine in the experiments related to the gastrointestinal digestion and absorption of nutrients and medicines.【Method】 A total of 11 nine-month-old Bama Minipigs (17.5 kg±2.0 kg) were selected for this study.After anesthesia,an incision was made in the right abdominal region to expose the intestines,followed by purse-string suturing.The cannulas were inserted and fixed,and the incisions were closed.Postoperative care was carefully managed to prevent infection.Once the animals had fully recovered,the experiment was conducted using direct ultra-high temperature (dUHT) milk as a test substance.Intestinal effluents and blood samples were analyzed using SDS-PAGE,confocal laser scanning microscopy (CLSM),and ultra-performance liquid chromatography (UPLC) to detect changes in protein and amino acid content.The dynamic changes in milk protein during digestion and absorption were assessed to confirm the reliability and applicability of the model for simulating food digestion and absorption.【Result】 All 11 experimental animals survived for two months post-surgery without any significant discomfort or complications,and showed good growth,with an average weight gain of 12.23 kg,consistent with normal growth trends.During the experimental period,no cannulas detachment was observed,and the cannulas functioned normally,enabling the smooth collection of duodenal and ileal effluents as well as blood samples.In this experiment,the protein microstructure of the effluent samples was examined,and it was observed that the protein content gradually decreased with the digestion time continuously increased.During digestion,casein and β-lactoglobulin bands showed a tendency of enhancing and then weakening.Detecting the amino acid content in the blood at different time points before and after ingestion,and the amino acid absorption curves were obtained.The incremental area under the curve (iAUC) of total amino acid in plasma was (12 105.00±2 145.57) μg·min/mL after ingestion,which was consistent with the normal kinetics of milk protein absorption.【Conclusion】 This study successfully established the double cannulas model involving the duodenum and the terminal ileum in Bama Minipigs,validating the effectiveness of the surgical technique and postoperative care regimen.The double cannulas model demonstrated good application prospects in Bama Minipigs,providing a reliable experimental animal model for studies on gastrointestinal digestion and absorption.This model offered strong support for exploring the digestion and absorption of nutrients,new drug development,and evaluating the absorption of existing drugs.

Key words: Bama Minipigs; double intestinal cannulas; digestive absorption; postoperative care

中图分类号:

S828

秦锡雨, 刘晓雪, 柴懿, 赖孟瑄, 任晓敏, 张德鹏, 张鹏, 李军涛, 李依璇, 王然, 郝彦玲, 武会娟, 王晓玉. 巴马小型猪十二指肠及回肠末端双瘘管模型构建及评价[J]. 中国畜牧兽医, 2025, 52(5): 2078-2087.

QIN Xiyu, LIU Xiaoxue, CHAI Yi, LAI Mengxuan, REN Xiaomin, ZHANG Depeng, ZHANG Peng, LIJuntao, LI Yixuan, WANG Ran, HAO Yanling, WU Huijuan, WANG Xiaoyu. Establishment and Evaluation of Double Cannulas Model of Duodenal and Terminal Ileum of Bama Minipigs[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(5): 2078-2087.

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