脾虚型咽喉反流性疾病中医病证结合大鼠模型的构建与评价  

Establishment and evaluation of TCM disease and syndrome combination model of laryngopharyngeal reflux disease in rats with spleen deficiency

作　　者：刘昱辛 杨斌[2] 王昭倩 高歌 刘卓颖 徐春英[1] LIU Yuxin;YANG Bin;WANG Zhaoqian;GAO Ge;LIU Zhuoying;XU Chunying(Department of Otolaryngology,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Department of Pathology,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091;Graduate School,China Academy of Chinese Medical Sciences,Beijing 100700)

机构地区：[1]中国中医科学院西苑医院耳鼻喉科,北京100091 [2]中国中医科学院西苑医院病理科,北京100091 [3]中国中医科学院研究生院,北京100700

出　　处：《北京中医药》2025年第3期331-336,共6页Beijing Journal of Traditional Chinese Medicine

基　　金：中国中医科学院科技创新工程重大攻关项目(CI2021A02701)。

摘　　要：目的构建脾虚型咽喉反流性疾病(LPRD)中医病证结合大鼠模型,并对模型进行评价。方法按随机数字表法将大鼠分为对照组、模型组各8只。模型组给予睡眠剥夺加高脂饮食,对照组给予正常睡眠加高脂饮食。观察大鼠一般情况,苏木精-伊红(HE)染色观察大鼠喉黏膜组织病理变化,电子显微镜观察喉黏膜超微结构。结果造模第8周,模型组大鼠口咽部pH测定结果表明,均出现不同程度的喉咽反流,提示LPRD造模成功。造模过程中模型组大鼠死亡1只,为反复溺水致肺部感染。与对照组比较,模型组大鼠“消瘦”明显,体质量低于对照组(P<0.05);皮毛稀疏、毛色枯槁、拉尾排便率高;抓力低(P<0.05),反应迟钝、对环境刺激反应减弱;舌面色彩分析的R值低(P<0.05),G、B值高,但差异无统计学意义(P>0.05)。模型组大鼠喉黏膜黏膜上皮纤毛断裂、缺失,纤毛柱状上皮形态不规则,局部呈扁平上皮,局部为纤维支架;黏膜下腺体管腔扩张,腔内分泌物积聚,全层可见不等量炎性细胞浸润,黏膜中出现“瘤”样改变;黏膜增厚明显,视野内均无法显示喉软骨。模型组大鼠喉部黏膜上皮电镜下表现:线粒体肿胀,大部分呈空泡改变,线粒体膜缺损,嵴断裂脱颗粒,内质网扩张,核糖体数目减少;黏膜紧密连接较对照组短,连接缝隙大,结构松散。结论长期睡眠缺乏和高脂饮食可导致大鼠脾虚型LPRD,大鼠喉组织出现声门下区组织病变,喉组织线粒体出现“脾虚”改变。推测反流事件影响了喉细胞内线粒体动力学和线粒体自噬等关键过程调控,喉细胞功能下降,喉细胞屏障(包括喉细胞间隙、细胞间缝隙连接、桥粒、黏附带等)功能下降,表现为黏膜紧密连接变短,连接缝隙大,结构松散,这些改变均形成LPRD易感因素。该方法操作简单,成功建立该疾病的中医病证结合模型。Objective To construct traditional Chinese medicine(TCM)disease and syndrome combination model of laryngopharyngeal reflux disease(LPRD)in rats with spleen deficiency and evaluate the model.Methods Rats were randomly divided into the control group and the model group,with 8 rats in each group.The model group was subjected to sleep deprivation and a highfat diet,while the control group received normal sleep and a high-fat diet.The general condition of the rats was observed,and hematoxylin and eosin(HE)staining was used to observe the pathological changes of the laryngeal mucosa.Electron microscopy was employed to observe the ultrastructure of the laryngeal mucosa.Results At the 8th week of modeling,pH measurements of the oropharynx in the model group showed varying degrees of laryngopharyngeal reflux,indicating the successful establishment of the LPRD model.During the modeling process,one rat in the model group died due to repeated drowning leading to a pulmonary infection.Compared with the control group,the model group showed significant"weight loss",with body weight lower than that of the control group(P<0.05).Fur was sparse,hair color was withered,and tail defecation rate was high.The grasping strength was low(P<0.05),and the rats exhibited slow responses with a reduced reaction to environmental stimuli.The R-value of tongue color analysis was lower in the model group(P<0.05),while the G and B values were higher,but the difference was not statistically significant(P>0.05).The laryngeal mucosa of model group showed ciliary breakage and loss in the epithelial layer,with irregular columnar epithelial morphology and areas showing squamous epithelium and fibrous scaffolds.The glandular lumens in the submucosa were dilated,with secretions accumulating inside,and inflammatory cell infiltration was observed throughout the mucosa,with tumor-like changes in some areas.The mucosa was significantly thickened,and the laryngeal cartilage was not visible in the field of view.Under electron microscopy,the laryngeal mucosal epi

关 键 词：咽喉反流性疾病 脾虚证 中医病证结合模型 线粒体自噬 大鼠 

分 类 号：R276.1[医药卫生—中医五官科学]