胰高血糖素样肽-2对28日龄断奶仔猪肠上皮细胞的保护及修复效应研究  

Cytoprotection and Repairing Effects of GLP-2 on Intestinal Epithelial Cells of 28-d Weaned Piglets Injuried by β-conglycinin in vitro

作　　者：贾刚[1] 蒋荣川[1] 晏家友[1] 黄兰[1] 王康宁[1] 

机构地区：[1]四川农业大学动物营养研究所教育部动物抗病营养重点实验室,雅安625014

出　　处：《畜牧兽医学报》2009年第10期1478-1486,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：教育部长江学者和创新团队发展计划"猪抗病营养的分子作用机制"(IRTO555);四川省应用基础研究"生物活性肽对仔猪断奶应激综合症的营养调控"(045Y029-031)

摘　　要：本研究旨在探讨胰高血糖素样-肽2(Glucagon-like peptide-2,GLP-2)对28日龄断奶仔猪的肠黏膜上皮细胞(IEC)损伤后增殖、代谢、凋亡的影响及可能的作用机理。试验首先采用单因子设计,分别用1.2和2.4 mg.mL-1的β-伴球蛋白对原代培养的断奶仔猪肠上皮细胞进行攻毒,通过测定细胞增殖、代谢及凋亡的变化而建立细胞损伤模型;然后再采用2×3因子设计,考察添加不同浓度的GLP-2(1×10-9、1×10-8、1×10-7mol.L-1)对致敏的肠上皮细胞的影响。结果表明,使用β-伴球蛋白攻毒,细胞MTT OD值显著降低(P<0.05),细胞蛋白质沉积量和细胞总蛋白含量极显著降低(P<0.01),胞外LDH活力极显著增加(P<0.01),Na+-K+-ATP酶活力显著(P<0.05)或者极显著(P<0.01)降低,半胱氨酸蛋白酶-3(caspase-3)酶活力极显著(P<0.01)升高;使用β-伴球蛋白攻毒的同时添加不同浓度的GLP-2,细胞MTT OD值、细胞蛋白质沉积量、细胞总蛋白含量和Na+-K+-ATP酶活力均显著或极显著(P<0.05或P<0.01)升高,且随着GLP-2浓度的增加而升高,而胞外LDH活力则随着GLP-2浓度的增加而逐渐下降(P<0.05或P<0.01),caspase-3酶活力极显著(P<0.01)降低。提示β-伴球蛋白对断奶仔猪小肠上皮细胞的增殖和细胞完整性有不利影响,而GLP-2能够减轻或者避免β-伴球蛋白对断奶仔猪小肠上皮细胞的不利影响,这种效应可能是通过调节细胞Na+-K+-ATP酶、caspase-3等的活力变化并影响细胞代谢而实现。The experiment was conducted to study the effects of GLP-2 on proliferation, metabo- lism and apoptosis of intestinal epithelial cells after injury by β-conglycinin in 28-day old weaned piglets and GLP-2＇s possible action mechanism. According to single-factor design principle, dif ferent β-conglycinin concentrations of 1.2 and 2.4 mg . mL^-1 were used for conteracting toxic substances to primary culture epithelial cells of weaned piglets. The cell damage model was established on the investigation of the cell proliferation, metabolism and apoptosis. Then the experiment of 2 × 3 factorial design was adopted to study the impacts of the sensitized cultured cell with different GLP-2 concentrations of 1 × 10^-9, 1 × 10^-8 and 1 × 10^-7mol.L^-1. The results were as fol lows： when β-conglycinin were used for counteracting toxic substances, the MTT OD significantly decreased （P〈0.05） ; the protein retention and total protein content in cell significantly decreased （P〈0.01） ; LDH activity significantly increased （P〈0.01） ; Na^＋-K^＋-ATP enzyme activity significantly decreased （P〈0.05）; caspase-3 activity significantly increased （P〈0.01）. When β-conglycinin were used for conteracting toxic substances after addition of GLP-2, the MTT OD, the protein retention, total protein content and Na^＋-K^＋-ATP enzyme activity significantly increased （P〈0. 05）. LDH activity gradually decreased （P〈0. 05 or P〈0. 01） and caspase-3 activity significantly decreased （P〈0.01） along with the increase of GLP-2 concentration. The results indicated that β-conglycinin had adverse effects on intestinal epithelial cells proliferation and cell integrality in vitro. GLP-2 could relieve or avoid the adverse effects on intestinal epithelial cells proliferation and cell integrality by regulating Na^＋-K^＋-ATP enzyme activity and caspase-3 activity, and consequently affecting cell metabolism.

关 键 词：胰高血糖素样肽-2 β-伴球蛋白 断奶仔猪 肠上皮细胞 增殖 代谢 

分 类 号：S828[农业科学—畜牧学] S816.7[农业科学—畜牧兽医]