猪血凝性脑脊髓炎病毒感染降低ZO-1表达致小鼠血脑屏障通透性升高

发布时间：2018-04-28 21:34

本文选题：猪血凝性脑脊髓炎病毒 + 血脑屏障　；参考：《中国兽医学报》2017年03期

【摘要】：为明确猪血凝性脑脊髓炎病毒(porcine hemagglutinating encephalomyelitis virus,PHEV)感染后对机体血脑屏障(blood-brain barrier,BBB)通透性的影响,本试验以PHEV易感的BALB/c小鼠为实验动物模型,将PHEV接种小鼠后静脉注射伊文氏蓝(Evans blue,EB),运用形态学和化学定量方法检测EB通过BBB进入脑组织的状况。结果显示,接种病毒后3d的小鼠脑组织即出现肉眼可见的蓝色,定量检测结果证实其含量随着感染时间的延长而增加。同时利用荧光定量PCR和Western blot方法分别对病毒感染后不同时间点紧密连接蛋白ZO-1、Occludin和Claudin-5进行核酸定量检测,证明感染组小鼠脑组织中ZO-1的基因转录水平随感染时间延长下调明显,而Occludin和Claudin-5的变化不明显。进而对脑组织中ZO-1蛋白表达水平进行检测,证实ZO-1的蛋白表达水平也发生下调,表明内皮细胞紧密连接完整性在病毒感染后期遭到了破坏。上述试验结果证实PHEV感染机体后可使BBB的通透性升高,而且这一变化与ZO-1的降低相关,为后期深入开展PHEV所致神经系统炎性损伤的机制奠定基础。
[Abstract]:In order to investigate the effect of porcine hemagglutinating encephalomyelitis virus (PHEV) infection on the permeability of blood-brain barrier (BBB) in porcine hemagglutinating encephalomyelitis virus, BALB/c mice susceptible to PHEV were used as experimental model. PHEV was injected intravenously into mice after inoculation with PHEV. The brain tissues of Epstein-Barr (EB) were detected by morphological and chemical quantitative methods. The results showed that the blue was visible in the brain tissue of mice 3 days after inoculation, and the quantitative test confirmed that the content of the virus increased with the prolongation of the time of infection. At the same time, fluorescence quantitative PCR and Western blot were used to detect the nucleic acid of ZO-1Occludin and Claudin-5 at different time points after infection, respectively. The results showed that the transcription level of ZO-1 gene in the brain tissue of the infected mice was down-regulated with the prolongation of infection time. However, the changes of Occludin and Claudin-5 were not obvious. Furthermore, the expression of ZO-1 protein in brain tissue was detected, which confirmed that the expression of ZO-1 protein was down-regulated, indicating that the integrity of tight junction of endothelial cells was damaged in the later stage of virus infection. The above results confirm that PHEV infection can increase the permeability of BBB, and this change is related to the decrease of ZO-1, which lays a foundation for the further development of the mechanism of PHEV induced inflammatory injury of nervous system.
【作者单位】：吉林大学动物医学学院;
【基金】：国家自然科学基金资助项目(31472194,31272530) 吉林省青年基金资助项目(20160520033JH) 吉林大学“大学生创新创业训练计划”资助项目(2014A81366)
【分类号】：S852.65

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