黄芪多糖对肠炎雏鸡小肠黏膜损伤的保护作用  

Protective Effect of Astragalus Polysaccharide on Intestinal Mucosal Injury in Chicks with Enteritis

作　　者：刘颖 田旭 冯晓梦 吕晓萍[1] 高雪丽[1] 郑世民[1] 刘超男[1] LIU Ying;TIAN Xu;FENG Xiaomeng;LYU Xiaoping;GAO Xueli;ZHENG Shimin;LIU Chaonan(Heilongjiang Key Laboratory for Laboratory Animals and Comparative Medicine,College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China;Pharmaron Beijing Co.,Ltd.,Beijing 102200,China)

机构地区：[1]东北农业大学动物医学学院,黑龙江省实验动物与比较医学重点实验室,哈尔滨150030 [2]康龙化成(北京)新药技术股份有限公司,北京102200

出　　处：《中国畜牧兽医》2023年第1期76-85,共10页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31472169);黑龙江省教育厅科学技术研究项目基金(12511029)。

摘　　要：【目的】探究黄芪多糖(Astragalus polysaccharide,APS)对雏鸡肠道形态和局部黏膜免疫的影响,阐明APS减轻肠炎雏鸡肠黏膜损伤的作用机制。【方法】在构建LPS诱导肠炎模型试验中,选取15只14日龄SPF雏鸡随机分为3组:对照组(Con)、低剂量脂多糖(lipopolysaccharide,LPS)模型组(DL)和高剂量脂多糖模型组(DH),每组5只。对照组灌喂生理盐水,DL和DH组分别灌喂1和2 mg/kg BW LPS,连续处理3 d,取小肠组织,采用HE染色法观察小肠病理变化,采用实时荧光定量PCR检测白介素-1β(interleukin 1β,IL-1β)和肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)mRNA表达量,筛选构建肠炎模型的最佳LPS剂量。在APS对肠黏膜损伤的保护试验中,将20只7日龄雏鸡分为4组:对照组(C)、脂多糖炎症组(L)、黄芪多糖组(A)和黄芪多糖抑制炎症组(A+L),每组5只。A和A+L组从7日龄到试验结束每天自由饮用APS溶液(1.0 g/L),C组在此期间自由饮水;L和A+L组雏鸡14日龄时灌喂筛选所得剂量的LPS,连续3 d。取各组雏鸡胸腺、脾脏、法氏囊和小肠组织,计算雏鸡免疫器官指数;采用HE染色法观察雏鸡小肠黏膜形态,糖原PAS染色法检测杯状细胞数量,实时荧光定量PCR检测咬合蛋白-1(Occludin-1)、闭合蛋白-1(Claudin-1)和闭合小环蛋白-1(ZO-1)mRNA表达量。【结果】在构建LPS诱导肠炎模型试验中,DL、DH组雏鸡小肠组织均出现肠黏膜固有层充血、肠绒毛损伤等现象,且IL-1β和TNF-αmRNA表达量显著高于对照组(P<0.05),因此确定雏鸡灌喂1 mg/kg BW LPS建立肠炎模型。在APS对肠黏膜损伤的保护试验中,与对照组相比,L组雏鸡小肠绒毛破碎,固有层充血,免疫器官指数显著下降(P<0.05),小肠隐窝深度显著升高(P<0.05),绒腺比(V/C)显著下降(P<0.05),杯状细胞数量显著减少(P<0.05),紧密连接蛋白Occludin-1、Claudin-1和ZO-1的mRNA表达量均显著下降(P<0.05);与L组相比,A+L组雏鸡免疫器官指数显著增加(P<0.05),【Objective】 The study was aimed to investigate the effects of Astragalus polysaccharide(APS) on the intestinal morphology and local mucosal immune in chicks, in order to clarify the mechanism of APS in reducing intestinal mucosal injury in chickens with enteritis.【Method】 Fifteen 14-day-old SPF-grade chicks were randomly divided into three groups: Control group(Con),low-dose lipopolysaccharide(LPS) model group(DL) and high-dose LPS model group(DH),5 chicks in each group.Control group was gavaged with normal saline, DLand DHgroups were gavaged with 1 and 2 mg/kg BW LPS,respectively.After 3 days of continuous treatment, the small intestine were taken.HE staining method was used to observe the pathological changes of small intestine, and Real-time quantitative PCR was used to detect the changes in the expression of interleutin 1β(IL-1β) and tumor necrosis factor α(TNF-α),and screen the best LPS dose for constructing LPS-induced enteritis model.In the protection test of APS against intestinal mucosal injury, twenty 7-day-old chicks were divided into four groups: Control group(C),LPS inflammation group(L),APS group(A) and APS inhibited inflammation group(A+L),5 chicks in each group.A and A+L groups were treated with APS(1.0 g/L) from 7-day-old to the end of this experiment, C group was fed water during this period.Chicks in L and A+L groups were given the selected optimal dose of LPS,after 3 consecutive days, the thymus, spleen, bursa of Fabricius and small intestine tissues were collected, the immune organ indexes of chicks were calculated in each group.HE staining was used to detect the pathological changes of chick small intestine, PAS staining was used to observe the number changes of goblet cells, Real-time quantitative PCR was used to detect the mRNA expression of tight junction proteins Occludin-1,Claudin-1 and ZO-1.【Result】 In the experiment of constructing LPS-induced enteritis model, the small intestinal mucosa lamina propria of chicks in DLand DHgroups were congested and intestinal villi wer

关 键 词：黄芪多糖(APS) 肠炎 黏膜免疫 杯状细胞 紧密连接蛋白 

分 类 号：S853.74[农业科学—临床兽医学]