柔嫩艾美耳球虫感染对雏鸡肠道紧密连接相关蛋白表达的影响  

Effects of Eimeria tenella challenge on the expression of tight junction related protein in broiler chicken gut

作　　者：张艳冰 许笑 古少鹏[1] 蔡建平[2,3] ZHANG Yan-bing;XU Xiao;GU Shao-peng;CAI Jian-ping(College of Animal Science and Veterinary Medicine,Shanxi Agricultural University,Jinzhou 030801,China;State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;Jiangsu Co-innovation Center for Prevention and Control of Animal Infectious Diseases and Zoonoses,Yangzhou 225009,China)

机构地区：[1]山西农业大学动物科技学院,山西晋中030801 [2]中国农业科学院,兰州兽医研究所,家畜疫病病原生物学国家重点实验室,甘肃省动物寄生虫病重点实验室,甘肃兰州730046 [3]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《中国兽医科学》2020年第6期746-752,共7页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2017YFD0500403)。

摘　　要：为研究柔嫩艾美耳球虫感染对雏鸡肠道黏膜上皮组织Occludin和Claudin-1表达的影响,给14日龄雏鸡经嗉囊接种2×10^4个柔嫩艾美耳球虫的孢子化卵囊,分别于感染后第0、4、7、14和21天采集小肠和盲肠样本。用荧光定量PCR和Western-blotting分析感染后不同时间点雏鸡肠道黏膜上皮组织Occludin和Claudin-1的表达水平变化。结果显示:在m RNA水平上,相较于第0天,感染后第4天盲肠Occludin表达量极显著上升(P<0.01),第7天则显著下降(P<0.01),第14和21天表达量逐渐回升,但均无显著差异(P>0.05);Claudin-1的表达量呈先下降后上升的变化趋势,但均无统计学差异(P>0.05)。在蛋白水平上,相较于感染后第0天,小肠和盲肠的Occludin均在感染后第4天显著上升(P<0.05),在感染后第7天表达量显著下降(P<0.05),第14和21天表达量则显著升高(P<0.01);盲肠Claudin-1的表达量在第7天显著下降(P<0.01),小肠Claudin-1在第21天则显著上升(P<0.01)。结果表明,球虫感染后的裂殖生殖阶段可能会引起机体的抗损伤反应,使得Occludin的表达量显著上升,感染后第7天雏鸡肠道紧密连接蛋白Occludin和Claudin-1的表达量下降,是肠上皮屏障受损的标志。To study the effects of Eimeria tenella challenge on the main tight junction protein Occludin and Claudin-1 expression dynamics in the intestinal mucosa(small intestine and cecum)of broiler chicken,14-day-old chickens were inoculated with 2×10^4 sporulated E.tenella oocysts through craw,and samples of small intestine and cecum were collected at 0 d,4 d,7 d,14 d and 21 d post-infection(pi).Real-time PCR and Western-blotting were used to analyze the expression of Occludin and Claudin-1 in the intestinal mucosal epithelial at different time points after infection.The transcriptional results showed that,compared with 0 d pi,the expression of cecum Occludin significantly increased at 4 d pi(P<0.01),and decreased significantly at 7 d pi(P<0.01),14 d and 21 d the expression gradually increased,but there was no significant difference(P>0.05).The expression of Claudin-1 showed a trend of decreased first and increased later,but there was no statistical difference(P>0.05).The dynamic trend of Occludin expression at the translation level is similar to the transcription level,but increased significantly at 14 d and 21 d pi(P<0.01).The expression of cecum Claudin-1 significantly decreased at 7 d pi(P<0.01),and small Claudin-1 significantly increased at 21 d pi(P<0.01).These results suggested that the challenge of E.tenella may cause the body anti-damage responses during its schizogonic stage,which makes the expression of Occludin significantly increase.Moreover,the significant decrease of Occludin and Claudin-1 at 7 d pi may be the sign of intestinal epithelial barrier injury.

关 键 词：柔嫩艾美耳球虫 雏鸡 肠上皮屏障 紧密连接 OCCLUDIN CLAUDIN-1 

分 类 号：S852.723[农业科学—基础兽医学]