伪狂犬病病毒感染小鼠基质金属蛋白酶-9介导紧密连接蛋白损伤血脑屏障的研究

doi:10.11843/j.issn.0366-6964.2024.05.036

摘要/Abstract

摘要： 旨在探究伪狂犬病病毒(pseudorabies virus，PRV)对小鼠血脑屏障(blood-brain barrier，BBB)通透性的影响及基质金属蛋白酶-9(matrix metalloproteinase-9，MMP-9)与紧密连接(tight junction，TJ)的相关性。将昆明SPF小鼠96只，随机平均分为对照组和3个试验组，24只对照组小鼠滴鼻生理盐水，72只试验组小鼠滴鼻感染PRV，将试验组分为感染24 h组、感染48 h组和感染72 h组，每组24只。观察临床症状并采用H&E染色观察病理损伤情况；通过改良型神经学评分和脑组织病毒载量测定评价PRV感染后神经损伤情况与病毒载量的关系；通过干湿重法和伊文斯蓝染色法探究血脑屏障的通透性；通过mRNA和蛋白表达差异水平探究MMP-9和TJ的相关性。结果显示，攻毒后脑组织病变程度随时间延长而加剧，在攻毒48 h后，脑组织病毒载量出现显著性差异；神经功能损伤评分在第72小时达到最高；干/湿重值及脑组织伊文斯蓝浓度呈时间依赖性；MMP-9与TJ的相关性结果表明，PRV攻毒后MMP-9表达量增加，TJ表达量减少。结果表明，PRV感染通过MMP-9介导的TJ蛋白表达量的降低损害BBB的完整性及通透性。综上所述，PRV感染可能通过MMP-9介导的破坏来损害BBB，为进一步阐明PRV入侵机制奠定基础。

关键词: MMP-9, 伪狂犬病病毒, 血脑屏障, TJ蛋白

Abstract: The experiment aims to investigate the effect of pseudorabies virus (PRV) on the permeability of blood-brain barrier in mice and the correlation between matrix metalloproteinase-9 (MMP-9) and tight junction (TJ). Ninety-six Kunming SPF mice were randomly divided into control group and experimental group. Twenty-four mice of control group received nasal saline and seventy-two mice of experimental group received nasal PRV infection. The experimental groups were divided into 24 h infection group, 48 h infection group and 72 h infection group, each group consisted of 24. The clinical symptoms and the pathological lesions were observed by H&E staining. The relationship between nerve damage and viral load after PRV infection was evaluated by modified neurological score and viral load measurement in brain tissue. The permeability of blood-brain barrier was investigated by wet-dry weight method and Evans blue staining method. The correlation between MMP-9 and TJ was explored through mRNA and protein expression differences. The results showed that the degree of brain lesions increased with the extension of time, and the nerve function injury score reached the highest at the 72nd hour. There were significant differences in viral load in brain tissue 48 hours after challenge. W/D, and Evans blue concentration in brain tissue were time-dependent. The correlation between MMP-9 and TJ showed a positive correlation between MMP-9 and TJ expression. After PRV infection, the expression of MMP-9 increased, and the expression of TJ decreased. The results showed that PRV infection impairs BBB integrity and permeability through an MMP-9-mediated decrease of TJ protein expression. In summary, PRV infection may damage BBB through MMP-9-mediated destruction, which lays a foundation for further elucidation of the PRV invasion mechanism.

Key words: MMP-9, pseudorabies virus, blood-brain barrier, TJ protein

中图分类号:

S858.285.3

张颖, 宋春莲, 张莹, 沈鸿, 舒相华, 杨洪贵. 伪狂犬病病毒感染小鼠基质金属蛋白酶-9介导紧密连接蛋白损伤血脑屏障的研究[J]. 畜牧兽医学报, 2024, 55(5): 2186-2194.

ZHANG Ying, SONG Chunlian, ZHANG Ying, SHEN Hong, SHU Xianghua, YANG Honggui. Study on the Damage of Blood-brain Barrier by Tight Junction Protein Mediated by MMP-9 in Pseudorabies Virus-infected Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2186-2194.

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