咽喉反流性疾病中SP、NK-1R、IL-8、PGP9.5的表达及意义  

Expression and Significance of SP,NK-1R,IL-8,PGP9.5 in Laryngopharyngeal Reflux Disease

作　　者：王刚 林春兰 刘蔓林 陈函 王茂鑫 陈贤明 Wang Gang;Lin Chunlan;Liu Manlin;Chen Han;Wang Maoxin;Chen Xianming(Department of Otolaryngology Head and Neck Surgery,the 900th Hospital of Joint Logistic Support Force,PLA,Fuzhou,350025,China)

机构地区：[1]中国人民解放军联勤保障部队第九〇〇医院耳鼻咽喉头颈外科,福州350025

出　　处：《听力学及言语疾病杂志》2022年第6期582-586,共5页Journal of Audiology and Speech Pathology

基　　金：福建省自然科学基金面上项目(2020J011143)。

摘　　要：目的初步探讨神经源性炎症与咽喉反流性疾病的关系,为咽喉反流性疾病的机制研究提供新思路。方法20只雄性豚鼠随机分为实验组和对照组,每组10只,均取头低脚高位,经口插入硬膜外麻醉管至食管中段,实验组灌注模拟胃液,对照组灌注生理盐水,以8滴/分速度灌注,每次20 min,灌注14天后取声门后联合区黏膜组织,行常规病理组织检查白细胞浸润程度;用蛋白印记法检测神经肽受体1(neuropeptide receptor-1,NK-1 R)、蛋白基因产物9.5(protein gene product 9.5,PGP 9.5)蛋白表达;用实时荧光定量PCR分别检测实验组和对照组P物质(substance P,SP)、NK-1R、白细胞介素8(IL-8)、PGP 9.5等mRNA表达,比较两组结果。结果①常规病理组织检查显示,实验组豚鼠咽喉部黏膜白细胞浸润程度(29.78±4.27个)较对照组(7.25±2.82个)增多,差异有统计学意义(P<0.05);②蛋白印迹法结果显示,实验组PGP 9.5的相对灰度值(1.88±0.22)明显高于对照组(1.26±0.28),差异有统计学意义(F=26.13,P<0.05);实验组NK1R的相对灰度值为0.29±0.05,明显高于对照组(0.21±0.06),差异有统计学意义(F=8.46,P<0.05);③实时荧光定量PCR结果显示,实验组和对照组豚鼠咽喉部黏膜SP mRNA表达下调(0.57倍),组间差异无统计学意义(P>0.05);实验组豚鼠咽喉部黏膜的NK-1R、IL-8、PGP 9.5 mRNA表达与对照组相比均上调(分别为6.32、3.34、2.27倍),差异均有统计学意义(均为P<0.05)。结论SP、NK-1R、IL-8、PGP 9.5可能在咽喉反流性疾病咽喉部高敏感性机理中发挥作用,该作用与神经源性炎症有关。Objective To study the relationship between neurogenic inflammation and laryngopharyngeal reflux disease,and provide new ideas for the mechanism of laryngopharyngeal reflux disease.Methods Twenty male guinea pigs were selected to construct animal models and randomly divided into experimental group and control group,with 10 in each group.The experimental group was perfused with simulated gastric juice,while the control group was perfused with normal saline at the rate of 8 drops/min for 20 min each time.Mucosal tissues in the posterior commissure were collected after 14 days of perfusion.Histopathology was performed to detect the degree of leukocyte infiltration.Western-blot analysis was used to detect neuropeptide receptor-1(NK-1R)and protein expression of protein gene product 9.5(PGP 9.5).The mRNA expressions of SP,NK-1R(Neuropeptide receptor-1),IL-8 and protein gene product 9.5(PGP 9.5)in experimental group and control group were detected by quantitative real-time PCR.Results①Histopathology showed that the degree of leucocytes infiltration in laryngopharyngeal mucosa of guinea pigs in experimental group was higher than that in control group(29.78±4.27 vs 7.25±2.82),and the difference was statistically significant(P<0.05).②Western blot results showed that the relative gray value of PGP 9.5 in the experimental group was 1.88±0.22,which was significantly higher than that in the control group(1.26±0.28),with statistical significance(F=26.13,P<0.05).The relative gray value of NK-1R in experimental group was 0.29±0.05,which was significantly higher than that in control group(0.21±0.06),and the difference was statistically significant(F=8.46,P<0.05).③Quantitative real-time PCR showed that SP mRNA expression in the larynx mucosa of guinea pigs in the experimental group and control group was down-regulated(0.57 times),and there was no statistical significance between groups(P>0.05).The mRNA expressions of NK-1R,IL-8 and PGP 9.5 in laryngopharyngeal mucosa of guinea pigs in experimental group were all up-regu

关 键 词：咽喉反流性疾病 神经源性炎症 神经肽受体-1 P物质 蛋白基因产物9.5 

分 类 号：R766.49[医药卫生—耳鼻咽喉科]