胃蛋白酶在不同pH条件下对咽喉黏膜上皮的作用  

The effects of pepsin on the mucosal epithelium of the laryngopharynx under different pH condition

作　　者：郑彦焱 颜冬惠 陈十燕 龚宏勋 陈贤明 ZHENG Yanyan;YAN Donghui;CHEN Shiyan;GONG Hongxun;CHEN Xianming(Department of Otolaryngology Head and Neck Surgery,900 Hospital of the Joint Logistics Team,Fuzhou 350025,China;Fuzong Clinical Medical College of Fujian Medical University,Fuzhou 350025,China)

机构地区：[1]联勤保障部队第九〇〇医院耳鼻咽喉头颈外科,福州350025 [2]福建医科大学研究生部,福建医科大学福总临床医学院,福州350025

出　　处：《听力学及言语疾病杂志》2025年第2期161-165,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：目的探讨酸性及非酸性条件下胃蛋白酶对咽喉部黏膜上皮的损伤作用,为难治性咽喉反流性疾病治疗提供新思路。方法30只健康豚鼠随机分为3组,每组10只:A组灌注胃蛋白酶+盐酸溶液(pH=2),B组灌注胃蛋白酶+蒸馏水(pH=7),C组灌注蒸馏水(pH=7)。灌注14 d后取环后区黏膜采用HE染色、免疫组化、透射电镜检测炎症细胞浸润程度、occludin和claudin-3蛋白表达、上皮细胞间隙增宽程度,电镜下分析细胞亚显微结构。结果①A组黏膜下白细胞计数28.61±7.10个,B组29.89±7.04个,C组7.81±1.40个,A、B两组高于C组(P<0.05);②与C组相比较,A组occludin和claudin-3蛋白表达均下调(P<0.05),B组occludin和claudin-3蛋白表达差异无统计学意义(P>0.05);③电镜下A组黏膜上皮细胞间隙增宽,线粒体肿胀、嵴退化;B组黏膜上皮细胞膜完好,细胞间隙无增宽,但胞质内线粒体肿胀、嵴退化;C组黏膜上皮细胞间隙无增宽,线粒体形态标志正常。结论与酸性条件下相比,非酸性条件下胃蛋白酶虽不引起细胞间隙增宽,但也可造成咽喉部黏膜上皮炎症性改变和线粒体损伤。Objective To investigate the damage of pepsin to the mucosal epithelium of the pharynx under acidic and non-acidic conditions,and to provide new ideas for the treatment of refractory laryngopharyngeal reflux diseases.Methods A total of 30 healthy guinea pigs were selected and randomly divided into 3 groups(with 10 in each group).The animals in group A were perfused with 0.3%pepsin+hydrochloric acid solution(pH=2),the animals in group B were perfused with 0.3%pepsin+sterilized distilled water(pH=7),and the animals in group C were perfused with sterilized distilled water(pH=7).After 14 days of perfusion,the mucosal tissues of the post-cyclic area were examined by HE staining,immunohistochemistry,and transmission electron microscopy to detect the degree of inflammatory cell infiltration,the expression of occludin and claudin-3 proteins,the degree of epithelial cell gap widening,and also to analyze the changes of cell submicroscopic structure under electron microscopy.Results①The submucosal leukocyte count was 28.61±7.10 in group A,29.89±7.04 in group B,and 7.81±1.40 in group C.Compared with group C,the leukocyte count was higher in group A and group B(P<0.05).②Compared with group C,both occludin and claudin-3 protein expression were downregulated in group A(P<0.05),compared with group C,the difference between occludin and claudin-3 protein expression in group B was not statistically significant(P>0.05).③Under electron microscopy,the mucosal epithelial cell gap was widened,the mitochondria was swollen and the cristae was degraded in group A.In group B,there was no widening of the mucosal epithelial cell gap,but the mitochondria in the cytoplasm was swollen and the cristae was degraded,and there was no widening of mucosal epithelial cell gap and the morphological markers of mitochondria were clear in group C.Conclusion Under non-acidic conditions,pepsin still can cause inflammatory changes and mitochondrial damage in the mucosal epithelium of the pharynx.

关 键 词：咽喉反流性疾病 胃蛋白酶 PH值 电镜 

分 类 号：R73[医药卫生—肿瘤]