γ-分泌酶抑制剂对卵清蛋白诱导的分泌性中耳炎大鼠中耳超微结构的影响  

Effect of γ-secretase inhibitor on middle ear ultrastructures in ovalbumin-mediated otitis media with effusion in rats

作　　者：陈沛维 赵春丽[2] 王丹妮[1] 杨劲松[1] 李颖[1] 赵守琴[1] Chen Peiwei;Zhao Chunli;Wang Danni;Yang Jinsong;Li Ying;Zhao Shouqin(Department of Otorhinolaryngology Head and Neck Surgery,Beijing Tongren Hospital,Capital Medical University,Beijing 100730,China;Department of Otorhinolaryngology Head and Neck Surgery,Beijing Youyi Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京同仁医院耳鼻咽喉头颈外科,100730 [2]首都医科大学附属北京友谊医院耳鼻咽喉头颈外科,100050

出　　处：《中华耳鼻咽喉头颈外科杂志》2021年第6期596-602,共7页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：国家自然科学基金(81770989);北京市自然科学基金(7202032)。

摘　　要：目的研究Notch信号通路抑制剂γ-分泌酶抑制剂DAPT对分泌性中耳炎(OME)大鼠中耳超微结构的影响。方法15只雄性SD大鼠完全随机化分组分为3组(对照组、OME组、OME+DAPT组),每组5只。OME组应用卵清蛋白(ovalbumin,OVA)全身及局部致敏建立OME模型,对照组以磷酸盐缓冲液(PBS)处理,OME+DAPT组在OME建模的基础上予以DAPT腹腔及鼓室注射。应用内镜、苏木精-伊红(HE)染色、扫描电镜比较各组中耳无纤毛区及纤毛区组织学及黏液-纤毛超微结构变化。采用单因素方差分析及Tukey法进行统计学分析。结果HE染色示对照组、OME组、OME+DAPT组在中耳黏膜非纤毛区和纤毛区黏膜下厚度差异均有统计学意义[非纤毛区:(6.83±1.47)μm比(38.58±9.57)μm比(32.17±11.89)μm,F=107.90;纤毛区:(26.69±3.22)μm比(30.41±6.75)μm比(26.76±4.06)μm,F=5.62;P值均<0.01]。OME组在非纤毛区和纤毛区的黏膜下厚度均较对照组明显增厚(F值为42.08和4.40,P值均<0.05),OME+DAPT组在非纤毛区与纤毛区的黏膜下厚度均较OME组减低,差异有统计学意义(F值为1.55和2.77,P值均<0.05)。扫描电镜观察纤毛区见OME组中耳黏膜纤毛排列明显紊乱、倒伏;OME+DAPT组纤毛形态排列较OME组好转,仍有部分倒伏。对照组、OME组、OME+DAPT组杯状细胞计数分别为(9.87±1.92)、(15.67±5.77)、(10.33±1.99)个,3组差异有统计学意义(F=11.43,P<0.01)。OME组杯状细胞计数明显高于对照组(F=9.00,P<0.01),OME+DAPT组杯状细胞数目较OME组减少,差异有统计学意义(F=8.41,P<0.01)。结论在OVA诱导的变态反应相关的OME中存在中耳局部超微结构变化,尤其是中耳纤毛区黏膜纤毛的超微结构改变,包括纤毛倒伏、紊乱,杯状细胞增多。DAPT可通过Notch信号通路缓解中耳黏液纤毛转运系统的形态学损伤,减少杯状细胞数量及高分泌状态从而调节OVA诱导的OME的局部变态反应。Objective To study the effect of the inhibitor of Notch signaling pathway-γ-secretase inhibitor DAPT on the ultrastructures of middle ear in the ovalbumin(OVA)-mediated allergic OME in vivo.Methods Male Sprague-Dawley(SD)rats,weighing 250-300 g,were completely and randomly divided into three groups(5 rats,10 ears in each group):(1)Control group(2)OME group(3)OME+DAPT group.Rats in the OME group underwent systemic and local sensitization by intraperitoneal and intratympanic injection of ovalbumin to make the model of OVA-induced OME.Rats in the control group were sensitized with PBS.On the basis of establishing the OME model,OME+DAPT group were intraperitoneal injected with DAPT(10 mg/kg)for seven consecutive days and were administered before intratympanic injection of ovalbumin.After the model was successfully established,endoscopy,H&E staining and scanning electron microscopy were used to study the histology and mucous-ciliary ultrastructures of the non-ciliated and ciliated mucosa in the middle ear of each group.One-way ANOVA and Tukey methods were used for statistical analysis.Results H&E staining showed that the three groups had statistically significant differences in submucosal thickness both in non-ciliated and ciliated regions(non-ciliated area:(6.83±1.47)μm,(38.58±9.57)μm,(32.17±11.89)μm,respectively.F=107.9;cilia area:(26.69±3.22)μm,(30.41±6.75)μm,(26.76±4.06)μm,respectively.F=5.62,both P<0.01).The thickness of the submucosa in the non-ciliated area and the cilia area of the OME group were significantly thicker than that of control group(F=42.08 and 4.40,both P<0.05);the thickness of the non-ciliated area and the ciliated area in OME+DAPT group were reduced compared to OME group(F=1.55 and 2.77,both P<0.05).Scanning electron microscopy showed that the array of cilia on the middle ear mucosa was disorderly arranged and inversed,this phenomenon was relieved in the OME+DAPT group.The number of goblet cells in the control group,OME group,and OME+DAPT group were 9.87±1.92;15.67±5.77;10.33±1.9

关 键 词：分泌性中耳炎 NOTCH信号通路 Γ-分泌酶抑制剂 超微结构 纤毛 

分 类 号：R764.21[医药卫生—耳鼻咽喉科]