在ALI模型中,山羊甲状腺流感病毒3型感染对山羊气道上皮质屏障功能的破坏
Lei-Lei Yang1, Min Sun1, Wen-Wen Zhang1
1Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences/Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture and Rural Affairs, Nanjing 210014, China; GuoTai (Taizhou) Center of Technology Innovation for Veterinary Biologicals, Taizhou 225300, China.
在空气-液体接口模型中的山羊气道上皮细胞被CPIV3感染,显示出障碍屏障功能受损. 确定NF-κB通路激活和细胞因子释放是破坏紧密结的关键机制.
科学领域:
- 呼吸系统病原体研究
- 表皮屏障的功能表皮屏障的功能
- 在体外建模模型.
背景情况:
- 呼吸上皮是利用粘膜清除和顶端结合综合体的关键防御屏障.
- 空气-液体接口 (ALI) 系统为研究气道上皮细胞特征提供了一个强大的体外模型.
研究的目的:
- 建立和描述ALI培养的山羊气道上皮细胞 (ALI-GAECs).
- 在ALI-GAEC中开发山羊肺炎病毒3 (CPIV3) 感染模型.
- 为了阐明CPIV3诱导的屏障功能障碍的机制.
主要方法:
- 建立和描述ALI-GAECs.
- 在ALI-GAEC中开发CPIV3感染模型.
- 使用TEER和FITC-Dextran透性的屏障功能的评估.
- 通过RT-qPCR和Western blot分析紧结蛋白表达 (ZO-1,ocludin,claudin-1) 的方法.
- 对信号通路 (NF-κB,MAPK) 和细胞因子表达 (TNF-α,IL-1β,IL-4,IL-6) 的研究.
- 对NF-κB通路的药理抑制.
主要成果:
- CPIV3感染导致ALI-GAEC的屏障功能受损,由降低TEER和增加透性证明.
- 病毒感染导致ZO-1和F-actin的重组.
- CPIV3降低了紧结蛋白的调节,并提高了对促炎性细胞因子的调节.
- NF-κB信号通路被激活,而MAPK通路没有受到显著影响.
- 抑制NF-κB部分恢复了紧结蛋白的表达,并降低了细胞因子水平.
结论:
- CPIV3感染通过激活NF-κB通路并诱导促炎性细胞因子的产生来破坏表皮屏障功能.
- ALI-GAECs模型是研究CPIV3病原体和其他山羊呼吸道病原体的宝贵工具.
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