TLR4/NF-κB信号通路在介导睡眠剥夺诱导膜迷路积水中的作用研究  

The Role of TLR4/NF-κB Signaling Pathway in Mediating Sleep Deprivation Induced Endolymphatic Hydrops

作　　者：贾玉其 胡涛 屈永涛[3] 许夏 郭明丽[3] Jia Yuqi;Hu Tao;Qu Yongtao;Xu Xia;Guo Mingli(Hebei Medical University,Shijiazhuang,050017,China)

机构地区：[1]河北医科大学,石家庄050017 [2]河北北方学院 [3]河北省人民医院耳鼻咽喉科

出　　处：《听力学及言语疾病杂志》2024年第4期342-348,共7页Journal of Audiology and Speech Pathology

基　　金：河北省政府资助临床医学优秀人才培养项目(2020002)。

摘　　要：目的探讨TLR4/NF-κB信号通路在介导睡眠剥夺诱导膜迷路积水中的作用。方法选取30只健康SD大鼠,分为空白对照组、大平台对照组、睡眠剥夺2周组、睡眠剥夺3周组、睡眠剥夺4周组,每组6只。睡眠剥夺组大鼠选用改良多平台睡眠剥夺法进行造模,大平台对照组在小平台上放置网格所制大平台,空白对照组不做处理。实验前后对各组大鼠进行旷场实验评估行为学,行听性脑干反应(ABR)检测评估听力水平;ABR检测结束后腹主动脉取血,通过酶联免疫吸附实验(ELISA)法检测血清中TNF-α、MCP-1的水平;解剖耳蜗,HE染色行中阶横截面积(SM)/(中阶+前庭阶横截面积)(SM+SV)比值评价膜迷路积水情况;通过免疫组化染色检测耳蜗中IL-1β、TNF-α、MCP-1、TLR4、NF-κB P65的表达,观察实验后不同睡眠剥夺时间大鼠听力水平、膜迷路积水程度及TLR4/NF-κB信号通路相关蛋白及下游炎症因子表达水平变化规律,分析大鼠听力水平、膜迷路积水程度与TLR4/NF-κB信号通路的相关性。结果睡眠剥夺组大鼠ABR波II反应阈升高,较空白对照组及大平台对照组均有统计学意义(P<0.01);形态学观察:空白对照组及大平台对照组积水率为0,睡眠剥夺2周组积水率为16.67%,睡眠剥夺3周组积水率为25%,睡眠剥夺4周组积水率为25%;睡眠剥夺组大鼠血清中TNF-α、MCP-1浓度均高于空白对照组及大平台对照组,其中睡眠剥夺4周组TNF-α、MCP-1浓度较空白对照组及大平台对照组均具有统计学意义;睡眠剥夺组大鼠耳蜗螺旋神经节、螺旋器、血管纹螺旋韧带IL-1β、TNF-α、MCP-1、TLR4、NF-κB P65表达均高于空白对照组及大平台对照组。结论睡眠剥夺可能通过TLR4/NF-κB信号通路对内耳产生影响从而诱导膜迷路积水。Objective To investigate the role of TLR4/NF-κB signaling pathway in mediating sleep deprivation induced endolymphatic hydrops.Methods A total of 30 healthy sprague-dawley(SD)rats were randomly divided into the control group、big platform control group,and 2 w,3 w,4 w sleep deprivation group,with 6 rats in each group.Modified multiple platform method was adopted to establish the rat sleep deprivation model.Before and after the experiment,the open field and acoustic brain-stem response(ABR)was conducted to evaluate the behavior and hearing level.After ABR test,blood samples were collected from abdominal aorta,and serum levels of TNF-αand MCP-1 were detected by ELISA.The cochlea was dissected,the severity of endolymphatic hydrops was assessed by calculating the ratio of the cross sectional area of the membranous cochlear duct(SM)to that of the membranous cochlear duct+scale vestibuli(SM+SV).Positive expression of IL-1β,TNF-α,MCP-1,TLR4,NF-κB P65 in rat cochlear tissues was detected via immunohistochemical staining.After the experiment,the changes of hearing level,the severity of endolymphatic hydrops and TLR4/NF-κB signaling pathway related proteins and downstream inflammatory factors expression level were observed.The correlation between TLR4/NF-κB signaling pathway and hearing level and endolymphatic hydropsin rats was analyzed.Results ABR results showed an increased threshold of waveⅡin the sleep deprivation group compared to those of the control group and big platform control group(P<0.05).The rate of hydrops was 0%in control and big platform control groups,16.67%in 2w sleep deprivation group and 25%in 3 w and 4 w sleep deprivation group.The concentrations of TNF-αand MCP-1 in serum of rats in sleep deprivation groups were higher than those in control and big platform control groups,and the 4w sleep deprivation group were statistically significant compared with control and big platform control groups.The expressions of IL-1β,TNF-α,MCP-1,TLR4 and NF-κB P65 in the cochlear spiral ganglion,spiral can

关 键 词：膜迷路积水 睡眠剥夺 TLR4/NF-κB信号通路 

分 类 号：R764.33[医药卫生—耳鼻咽喉科]