摘要
紧密连接在维持肠道黏膜完整性和免疫系统正常发挥功能中作用重大。【目的】旨在探讨猪圆环病毒2型(PCV2)感染对猪小肠上皮细胞(IPEC)紧密连接的影响。【方法】利用基因克隆获得Claudin-1蛋白和闭锁小带蛋白-1 (ZO-1)基因的重组质粒并建立荧光定量RT-PCR标准曲线;利用所建立的方法检测PCV2感染的IPEC中Claudin-1蛋白和闭锁小带蛋白-1mRNA的动态变化。【结果】构建的两种紧密连接蛋白基因的重组质粒荧光定量RT-PCR标准曲线线性关系良好,相关系数均达0.99以上。PCV2感染的IPEC中Claudin-1蛋白和ZO-1mRNA表达在4、12、24、48、72h均显著下降。【结论】PCV2感染可抑制猪小肠上皮细胞紧密连接蛋白相关基因的表达,破坏细胞间的紧密连接,增加肠道通透性。
Tight junctions play an important role in maintaining the integrity of intestinal mucosa and the normal functioning of the immune system.【Objective】The aim of this study was to investigate the effect of PCV2infection on tight junction of intestinal porcine epithelial cells(IPEC).【Methods】the recombinant plasmids of Claudin-1and Zonula Occluden-1(ZO-1)gene were obtained by gene cloning and their standard curves of fluorescence quantitative(FQ)RT-PCR were established.The dynamic changes of Claudin-1and ZO-1mRNA in IPEC infected with PCV2were detected by the method established.【Results】The standard curves of the recombinant plasmid(FQ)RT-PCR of the two tight junction proteins were linear,and their correlation coefficients were above 0.99.The expression of Claudin-1and ZO-1mRNA in PCV2infected IPEC decreased significantly at 4,12,24,48and 72hafter infection.【Conclusion】PCV2infection can inhibit the expression of tight junction protein related genes in IPECs,which maybe destroy the tight junctions between cells,and increase intestinal permeability.
作者
杨昕坦
阮峥
李焕荣
YANG Xintan;RUAN Zheng;LI Huanrong(Animal Science and Technology College,Beijing University of Agriculture,Beijing 102206,China)
出处
《北京农学院学报》
2019年第1期56-60,共5页
Journal of Beijing University of Agriculture
基金
2017年北京农学院大学生科研训练项目(5056516007/003)
