睡眠剥夺大鼠喉上皮细胞屏障功能改变在咽喉反流发病机制中的作用

The Role of Barrier Function of Laryngeal Epithelial Cells in the Pathogenesis of Laryngopharyngeal Reflux in Sleep Deprived Rats

郝梅;蒋兴旺;陈浩;尹龙龙;黎立;张延平;

1:河北北方学院

2:中国人民解放军总医院第八医学中心耳鼻咽喉头颈外科

3:中国人民解放军总医院第六医学中心耳鼻咽喉头颈外科医学部

4:国家耳鼻咽喉疾病临床医学中心

摘要
目的 探讨睡眠剥夺大鼠喉组织闭锁小带蛋白1(zonula occludens 1,Zo-1)及闭合蛋白3(claudin-3)表达变化,为揭示咽喉反流性疾病的发病机制提供参考。方法 健康成年雄性SD大鼠40只,随机分为实验组(30只),分为睡眠剥夺1月组、睡眠剥夺2月组和睡眠剥夺3月组,实验组使用改良多平台水环境法每天剥夺动物白天睡眠8 h,分别持续1、2和3月(分别为睡眠剥夺1、2、3月组,每组10只),对照组鼠笼饲养,正常睡眠。完成造模后各组分别处死取喉组织行Western blot和实时荧光定量RT-PCR分析,观察Zo-1及claudin-3蛋白和mRNA表达变化,比较四组的差异。结果 (1)与对照组比较,睡眠剥夺1月组、3月组Zo-1及claudin-3蛋白表达均降低,2月组Zo-1及claudin-3蛋白表达均升高,但差异无统计学意义(P>0.05);与2月组相比,3月组Zo-1蛋白表达下降(P<0.05),1月组及3月组claudin-3蛋白下降(P<0.05);(2)与对照组比较,1月组及2月组Zo-1 mRNA表达均上升,3月组Zo-1 mRNA表达下降,1月组claudin-3 mRNA表达升高,2月组及3月组claudin-3 mRNA表达降低,但差异无统计学意义(P>0.05);与1月组相比,3月组Zo-1 mRNA表达下降(P<0.05),2月组及3月组claudin-3 mRNA表达下降(P<0.05)。睡眠剥夺1、2、3月组之间比较,随着睡眠剥夺时间的延长,Zo-1和claudin-3蛋白表达先降低、后升高、然后再降低(P<0.05);Zo-1和claudin-3 mRNA表达逐渐降低(P<0.05)。结论 睡眠剥夺可能通过氧化应激干扰大鼠喉粘膜上皮主要紧密连接蛋白的表达,导致喉上皮细胞屏障功能障碍,使反流物更容易进入细胞间隙,与咽喉反流的发生和发展密切相关。
关键词
睡眠剥夺;紧密连接;闭锁小带蛋白1;闭合蛋白3;咽喉反流
基金项目(Foundation):
北京市自然科学基金面上项目(7192202)
作者
郝梅;蒋兴旺;陈浩;尹龙龙;黎立;张延平;
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