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The specific role of the tight junction proteins occludin and claudin-5 on the blood-brain barrier during Listeria monocytogenes infection

机译:紧密结蛋白的特定作用occludin和克劳丁-5在李斯特菌单核细胞增生期间血脑屏障感染

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摘要

To investigate the blood-brain barrier (BBB) permeability of mice after Listeria monocytogenes infection for further study on the mechanism of L. monocytogenes crossing the BBB, a mouse model was established and Evans blue assay was performed to assess the BBB disruption. Using relative quantitative real-time PCR, the RNA expression of Zonula occludens-1 (ZO-1), occludin and claudin-5 were detected. In addition, the protein expression level of ZO-1, occludin and claudin-5 were detected by immunohistochemistry and western blot. The extravasation of Evans blue dye was significantly different between 24 h and 96 h (P < 0.05). The mRNA expression of occludin and claudin-5 were down-regulated than that of the control group at each sampling point (P < 0.05) and ZO-1 showed a significant change at 96 h (P < 0.05). In addition, the protein expression level of occludin and claudin-5 decreased significantly at 48 h and 96 h (P < 0.05) by immunohistochemistry and western blot, compared with the control, while ZO-1 was almost unchanged (P > 0.05). All results indicating that the tight junction integrity of endothelial cells was destroyed and BBB permeability was enhanced in the process of L. monocytogenes infection, and this change was related to the decrease of the expression occludin and claudin-5.
机译:为了探讨小鼠的血脑屏障(BBB)小鼠在李斯特菌单核细胞增生后进行进一步研究L.单核细胞增生的机理的进一步研究,建立小鼠模型,并进行了emans蓝色测定以评估BBB破坏。使用相对定量的实时PCR,检测Zonula occludens-1(ZO-1),occludin和克劳德蛋白-5的RNA表达。此外,通过免疫组织化学和Western印迹检测ZO-1,occludin和Claudin-5的蛋白表达水平。 Evans Blue Dye的外向在24小时和96小时之间显着差异(P <0.05)。闭塞素和克劳丁蛋白-5的mRNA表达比每个取样点(P <0.05)的对照组的mRNA表达(P <0.05),ZO-1显示出96小时的显着变化(P <0.05)。此外,与对照相比,免疫组织化学和蛋白质印迹,Occludin和Claudin-5的蛋白表达水平在48小时和96小时(P <0.05)下降,而ZO-1几乎保持不变(P> 0.05)。所有结果表明,在L.单核细胞增生感染过程中,增加了内皮细胞的紧密结合的完整性,并且增强了BBB渗透性,并且这种变化与表达occludin和Claudin-5的减少有关。

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