紧密连接蛋白在声带白斑伴咽喉反流患者声带黏膜中的表达

摘要

Objective To study the roles of tight junction proteins (TJPs) in vocal mucosal barrier damage via probing their differential expressions in vocal mucosa of patients suffering from vocal leukoplakia accompanied with laryngopharyngeal reflux (LPR).Methods 26 patients suffering from vocal leukoplakia were divided into two groups (i.e.,groups with and without LPR) (13 cases in each group) according to the results of 24-h esophageal pH monitoring.Mucosal specimens of vocal cord were collected from all the patients during operation.HE staining was used to score the pathological changes of the mucosa.Transmission electron microscope (TEM) was utilized to observe the morphology of mucosal tight junction,and Western blot to detect the expressions of Claudin-1 and ZO-1.Results Compared with the group without LPR,marked inflammatory cell infiltration and poor structural integrity of the epithelium were observed via HE staining,TEM demonstrated significant decrease of electron dense materials around the tight junction and obvious morphological changes of the tight junction,and Western blot showed decreased expressions of Claudin-1 and ZO-1 in the specimens from the group with LPR.Furthermore,immunofluorescence assay confirumed that expressions of Claudin-1 and ZO-1 were decreased within cytoplasm in those from the group with LPR.Conclusion Refluxed materials may cause structural damage of tight junction and abnormal expressions of TJPs,thus leading to structural damage of intercellular junction,intercellular spaces dilatation and enhancement of epithelial permeability,which may be an important mechanism of vocal mucosal barrier damage in vocal leukoplakia accompanied with LPR.%目的 通过观察紧密连接(tight junction,TJ)蛋白在声带白斑伴咽喉反流(laryngophayngeal reflux,LPR)患者声带黏膜表达的差异,探讨其与声带黏膜屏障损伤的相关性.方法 将因声带白斑行手术治疗且资料完整的26例患者按24h双通道测酸结果分LPR组和非LPR组,两组各13例.术中分别采集声带黏膜标本.HE染色观察声带组织病理学的变化、透射电镜检测声带黏膜紧密连接的形态学、Western blot检测紧密连接蛋白Clau-din 1与ZO-1的表达.结果 HE染色发现LPR组声带黏膜炎症细胞浸润较多,上皮结构完整性较差;电镜下观察到LPR组紧密连接附近的电子致密物质明显较少,紧密连接形态学明显改变.Western blot示LPR组Claudin-1和ZO-1两种蛋白表达减少(P＜0.05).利用免疫荧光检测发现,LPR组的紧密连接蛋白Claudin-1和ZO-1在细胞浆内表达明显减少(P＜0.05).结论 咽喉反流物可以导致紧密连接结构破坏,相关蛋白功能表达异常,细胞间连接结构遭到破坏,细胞间隙扩大,声带黏膜上皮的通透性增高,可能是声带白斑黏膜损伤的重要机制.