金嗓散结胶囊对反流性咽喉炎模型大鼠咽喉黏膜屏障保护作用实验研究

Protective effect and related mechanism of Jinsang Sanjie capsules on throat mucosal barrier in reflux pharyngolaryngitis model rats

目的通过复制反流性咽喉炎大鼠模型,观察金嗓散结胶囊对反流性咽喉炎的作用。方法对大鼠进行切除下食道括约肌手术进行造模,12周后,将造模组大鼠分为5组,即模型组,金嗓散结胶囊高剂量组、中剂量组、低剂量组,阳性组,各组大鼠给予相应剂量药物经口灌胃治疗4周,进行反流体征评分,ELISA法检测大鼠血清白细胞介素(IL)-6、肿瘤坏死因子(TNF)-α、IL-1β、IL-10、IL-13含量;HE染色检测大鼠声带和食道组织病理变化;免疫荧光、RT-PCR技术以及Western blot法检测声带和食道组织紧密连接蛋白Claudin-3、Occludin、ZO-1的基因和蛋白表达。结果与模型组比较,金嗓散结胶囊可以减轻模型大鼠的声音沙哑,喉部等症状,降低反流体征评分,减轻声带和食道的病理,显著降低模型大鼠血清中IL-6、TNF-α、IL-1β的含量,上调IL-10、IL-13的含量,并显著增加模型大鼠Claudin-3、Occludin、ZO-1蛋白及其基因表达。结论金嗓散结胶囊治疗大鼠反流性咽喉炎可能通过抑制大鼠血清中炎症因子,上调血清抑炎因子,增强声带和食道中紧密连接蛋白表达,从而减轻反流性咽喉炎症状,修复咽喉黏膜的屏障系统,值得进一步研究。

Objective To determine the effect and mechanism of Jinsang Sanjie capsules on reflux pharyngolaryngitis rat model by replicating the rat model of reflux pharyngolaryngitis.Methods Rats models was estalished by removing the underwent lower esophageal sphincter.After 12 weeks,the model rats were divided into five groups,namely a model group,a high-dose,a medium-dose,a lowdose Jinsang Sanjie capsule group,and a positive group.The rats were given corresponding doses of drugs orally for 4 weeks,and the reflux signs were scored.ELISA was used to detect the serum content of interleukin(IL)-6,tumor necrosis factor-α(TNF-α),IL^(-1)β,IL^(-1)0,and IL^(-1)3 contents.HE staining was used to detect the histopathological changes in the vocal cords and esophageal tissue.Immunofluorescence,RT-PCR and Western blot were used to detect the gene and protein expression of tight junction proteins Claudin-3,Occludin and ZO-1 in the vocal cords and esophageal tissue.Results Compared with the model group,Jinsang Sanjie capsules alleviated symptoms such as hoarseness and throat discomfort in model rats,lowed reflux sign scores,alleviated the pathological symptoms of the vocal cords and esophagus,and reduced the content of IL-6,TNF-αand IL^(-1)β,increased the content of IL^(-1)0 and IL^(-1)3,which significantly increased Claudin-3,Occludin,and ZO-1 protein and gene expression of the model rats.Conclusion The mechanism of action of Jinsang Sanjie capsules for reflux pharyngolaryngitis in rats may be through inhibiting inflammatory factors in the rat serum,upregulating serum anti-inflammatory factors,and enhancing the expression of tight junctions in the vocal cords and esophagus to alleviate the symptoms of reflux pharyngolaryngitis.Repairing the barrier system of the throat mucosa needs further study.