腹水综合征肉鸡空肠黏膜组织结构及相关基因表达分析

doi:10.16431/j.cnki.1671-7236.2019.06.031

摘要/Abstract

摘要：

试验旨在研究肉鸡腹水综合征发生发展过程中空肠核因子κB （NF-κB）及紧密连接蛋白（闭合蛋白（occludin）、紧密连接蛋白-1（claudin-1）和胞质紧密黏连蛋白1（zonula occluden 1，ZO-1）） mRNA相对表达量，以及绒毛长度、隐窝深度和绒隐比的变化，为肉鸡腹水综合征的防治提供理论依据。选取40羽1日龄罗斯肉鸡常规饲养7 d后，随机分为对照组和模型组（饲料添加3%猪油和4%鱼粉，饮水添加0.12% NaCl，9~11℃低温饲养），35日龄检测体重，并取心脏组织测腹水心脏指数，取空肠组织以HE染色切片测量绒毛长度和隐窝深度，实时荧光定量PCR检测空肠NF-κB和紧密连接蛋白的基因表达量。结果表明，与对照组相比，模型组肉鸡体重极显著下降（P<0.01），腹水心脏指数极显著升高（P<0.01），空肠绒毛长度、绒隐比极显著下降（P<0.01），隐窝深度极显著升高（P<0.01），NF-κB mRNA相对表达量极显著升高（P<0.01），occludin、claudin-1和ZO-1 mRNA相对表达量极显著下降（P<0.01）。表明肠道黏膜结构与功能异常及肠通透性增高促进了肉鸡腹水综合征的发生发展。

关键词: 肉鸡腹水综合征; 空肠; 肠绒毛; 紧密连接蛋白

Abstract:

This experiment was aimed to study the relative expression of NF-κB and the tight junction protein(occludin,claudin-1 and ZO-1) mRNA and the changes of villus length (V),crypt depth (C) and V/C value in the development of broiler ascites syndrome,providing a theoretical basis for the prevention and treatment of broiler ascites syndrome.A total of 40 1-day-old Ross broilers were routinely reared for 7 days and randomly divided into control group and model group (added 3% lard and 4% fish meal to standard,added 0.12% NaCl to drinking water,and raising temperature at 9-11℃),and measured body weight at 35 days old.The ascites heart index was measured,and the jejunum tissue was taken for HE staining to measure the villus length and crypt depth.The gene expression of jejunal NF-κB and tight junction protein were detected by Real-time PCR.The results showed that compared with control group,the weight of broiler in model group was extremely significantly decreased (P<0.01),the ascites heart index was extremely significantly increased (P<0.01),the jejunum length and V/C value were extremely significantly decreased (P<0.01),the crypt depth was extremely significantly increased (P<0.01),the relative expression of NF-κB mRNA was extremely significantly increased (P<0.01),and the relative expression of tight junction proteins occludin,claudin-1 and ZO-1 mRNA were extremely significantly decreased (P<0.01).This results indicated that the intestinal mucosal structure and function abnormality and intestinal permeability increase promoted the development of broiler ascites syndrome.

Key words: broiler ascites syndrome; jejunum; intestinal villi; tight junction protein

中图分类号:

S856.5

温爽, 郭宇荣, 杨澜, 赵冬冬, 王卓, 申伟祺, 程佳, 孙耀贵, 王文魁, 李宏全, 段智变. 腹水综合征肉鸡空肠黏膜组织结构及相关基因表达分析[J]. 《中国畜牧兽医》, 2019, 46(6): 1825-1831.

WEN Shuang, GUO Yurong, YANG Lan, ZHAO Dongdong, WANG Zhuo, SHEN Weiqi, CHENG Jia, SUN Yaogui, WANG Wenkui, LI Hongquan, DUAN Zhibian. Analysis of Histological Structure and Expression of Related Genes in Jejunum Mucosa of Broilers Ascites Syndrome[J]. , 2019, 46(6): 1825-1831.

参考文献

[1] CURRIE R J.Ascites in poultry:Recent investigations[J].Avian Pathology,1999,28(4):313-326.
[2] 齐永华,王文魁.肉鸡腹水综合征研究进展[J].动物医学进展,2003,3:54-57. QI Y H,WANG W K.The progress of study on ascites syndrome in broilers[J].Progress in Veterinary Medicine,2003,3:54-57.(in Chinese)
[3] 韩烨晨.复方中药对腹水综合征肉鸡肝组织纤维化的影响[D].太谷:山西农业大学,2017. HAN Y C.Effect of the compound recipe of traditional Chinese medicine on liver fibrosis of broilers with ascites syndrome[D].Taigu:Shanxi Agricultural University,2017.(in Chinese)
[4] BALOG J M,ANTHONY N B,WALL C W,et al.Effect of a urease inhibitor and ceiling fans on ascites in broilers.2.Blood variables,ascites scores,and body and organ weights[J].Poultry Science,1994,73(6):810.
[5] CHEN C,CHEN R P,LIN H H,et al.Tolvaptan regulates aquaporin-2 and fecal water in cirrhotic rats with ascites[J].World Journal of Gastroenterology,2016,22(12):3363-3371.
[6] SAKAI H,SAGARA A,MATSUMOTO K,et al.5-fluorouracil induces diarrhea with changes in the expression of inflammatory cytokines and aquaporins in mouse intestines[J].PLoS One,2013,8(1):e54788.
[7] 韩烨晨,苏晓菲,王俊武,等.复方中药对肉鸡腹水综合征肺组织NO、ET-1的作用及机制[J].畜牧兽医学报,2015,46(6):1055-1062. HAN Y C,SU X F,WANG J W,et al.The effect and mechanism of compound recipe of the traditional Chinese medicine on NO,ET-1 in lung of broiler with ascites syndrome[J].Acta Veterinaria et Zootechnica Sinica,2015,46(6):1055-1062.(in Chinese)
[8] 张娟,王俊武,韩烨晨,等.复方中药对肉鸡腹水综合征肺组织血管内皮细胞源性增殖因子的影响[J].畜牧兽医学报,2017,48(1):174-184. ZHANG J,WANG J W,HAN Y C,et al.The effect of compound traditional Chinese medicine on endothelium derives multiplication factor in lung of broilers with ascites syndrome[J]. Acta Veterinaria et Zootechnica Sinica,2017,48(1):174-184.(in Chinese)
[9] LI K,ZHAO L,GENG G,et al.Increased calcium deposits and decreased Ca²⁺-ATPase in erythrocytes of ascitic broiler chickens[J].Research in Veterinary Science,2011,90(3):468-473.
[10] JULIAN R J,CASTON L J,MIRSALIMI S M,et al.Effect of poultry by-product meal on pulmonary hypertension,right ventricular failure and ascites in broiler chickens[J].The Canadian Veterinary Journal,1992,33(6):382-385.
[11] 张志浩.氧化应激对肉鸡肠道粘膜屏障功能的影响以及缓解肠道氧化损伤物质的研究[D].泰安:山东农业大学,2014. ZHANG Z H.Effects of oxidative stress on the intestinal barrier function of broiler chickens and the research of substance to relieve the injury[D].Tai'an:Shandong Agricultural University,2014.(in Chinese)
[12] 郑轲.脲酶抑制剂和脲酶卵黄抗体对肉鸡腹水症影响的研究[D].雅安:四川农业大学,2006. ZHENG K.Effects of urease inhibitor and anti-urease immunoglobulin Y on ascites in broilers[D].Ya'an:Sichuan Agricultural University,2006.(in Chinese)
[13] SOLIS D L S F,FARNELL M B,TELLEZ G,et al.Effect of prebiotic on gut development and ascites incidence of broilers reared in a hypoxic environment[J].Poultry Science,2005,84(7):1092-1100.
[14] ABDULKARIMI R,SHAHIR M H,DANESHYAR M.Effects of dietary glutamine and arginine supplementation on performance,intestinal morphology and ascites mortality in broiler chickens reared under cold environment[J].Asian Australasian Journal of Animal Sciences,2019,32(1):110-117.
[15] KARHAUSEN J,FURUTA G T,TOMASZEWSKI J E,et al.Epithelial hypoxia-inducible factor-1 is protective in murine experimental colitis[J].Journal of Clinical Investigation,2004,114(8):1098-1106.
[16] YEN J T,NIENABER J A,HILL D A,et al.Oxygen consumption by portal vein-drained organs and by whole animal in conscious growing swine[J].Proceedings of the Society for Experimental Biology Medicine,1989,190(4):393-398.
[17] 丁博文.大黄素对肠粘膜屏障缺氧/复氧损伤的保护作用及机理研究[D].南京:南京大学,2012. DING B W.The protective effect of emodin on intestinal mucosal barrier injury induced by hypoxia/reoxygenation and the investigation of its mechanism[D].Nanjing:Nanjing University,2012.(in Chinese)
[18] SONIS S T.The biologic role for nuclear factor-kappaB in disease and its potential invol vement in mucosal injury associated with anti-neoplastic therapy[J].Critical Reviews in Oral Biology & Medicine,2002,13(5):380-389.
[19] LORENZA G M,TAPIA R,CHAMORRO D.Crosstalk of tight junction components with signaling pathways[J].Biochimica et Biophysica Acta-Biomembranes,2008,1778(3):729-756.
[20] 高志光,秦环龙.肠上皮细胞紧密连接的生物学功能及在肠屏障中的作用[J].肠外与肠内营养,2005,12(5):299-302. GAO Z G,QIN H L.The biological functions of intestinal epithelial tight junctions and its role in intestinal barrier[J].Parenteral & Enteral Nutrition,2005,12(5):299-302.(in Chinese)
[21] CUMMINS E P,KEOGH C E,CREAN D,et al.The role of HIF in immunity and inflammation[J].Molecular Aspects of Medicine,2016,47-48:24-34.
[22] KASTHURI N,PREMILA A,REKHA K,et al.NF-κB-iNOS-COX2-TNFα inflammatory signaling pathway plays an important role in methotrexate induced small intestinal injury in rats[J].Food and Chemical Toxicology,2018,118:766-783.
[23] LUO H,GUO P,ZHOU Q.Role of TLR4/NF-kappaB in damage to intestinal mucosa barrier function and bacterial translocation in rats exposed to hypoxia[J].PLoS One,2012,7(10):e46291.
[24] ZAHRAOUI A,LOUVARD D,GALLI T.Tight junction,a platform for trafficking and signaling protein complexes[J].Journal of Cell Biology,2000,151(5):F31-F36.
[25] MITIC L L,VAN ITALLIE C M,ANDERSON J M.Molecular physiology and pathophysiology of tight junctions Ⅰ.Tight junction structure and function:Lessons from mutant animals and proteins[J].American Journal of Physiology-Gastrointestinal and Liver Physiology,2000,279(2):G250-G254.
[26] SUZUKI T.Regulation of intestinal epithelial permeability by tight junctions[J].Cellular and Molecular Life Sciences,2013,70(4):631-659.
[27] 黄宏春.肝硬化自发性腹水感染肠粘膜通透性改变的临床研究[D].郑州:郑州大学,2003. HUANG H C.A clinical practice study:Intestinal permeability changing in cirrhotic patients with spontaneous ascitic fluid infection[D].Zhengzhou:Zhengzhou University,2003.(in Chinese)
[28] UBEDA M,LARIO M,MUNOZ L,et al.Obeticholic acid reduces bacterial translocation and inhibits intestinal inflammation in cirrhotic rats[J].Journal of Hepatology,2016,64(5):1049-1057.
[29] LUO L,ZHOU Z,XUE J,et al.Bletilla striata polysaccharide has a protective effect on intestinal epithelial barrier disruption in TAA-induced cirrhotic rats[J].Experimental and Therapeutic Medicine,2018,16(3):1715-1722.
[30] ZHAO T Y,SU L P,MA C Y,et al.IGF-1 decreases portal vein endotoxin via regulating intestinal tight junctions and plays a role in attenuating portal hypertension of cirrhotic rats[J].BMC Gastroenterology,2015,15:77.
[31] SANCHEZ E,NIETO J C,BOULLOSA A,et al.VSL#3 probiotic treatment decreases bacterial translocation in rats with carbon tetrachloride-induced cirrhosis[J].Liver International,2015,35(3):735-745.
[32] ASSIMAKOPOULOS S F,TSAMANDAS A C,TSIAOUSSIS G I,et al.Altered intestinal tight junctions' expression in patients with liver cirrhosis:A pathogenetic mechanism of intestinal hyperpermeability[J].European Journal Clinical Investigation,2012,42(4):439-446.