摘要
目的基于水通道蛋白5(AQP5)-Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核转录因子κB(NF-κB)通路探讨桉柠蒎肠溶软胶囊对小鼠分泌性中耳炎(OME)模型的治疗作用及机制。方法昆明小鼠随机分为对照组、模型组、地塞米松组和桉柠蒎肠溶软胶囊低、中、高剂量(55、110、220 mg·kg^(-1))组。采用经鼓膜注入脂多糖(LPS)的方法建立小鼠OME模型,显微镜观察各组小鼠鼓膜结构变化,苏木精-伊红(HE)染色法评估中耳组织病理学改变,ELISA法检测中耳灌洗液中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平,Western blotting检测中耳黏膜组织AQP5及TLR4/MyD88/NF-κB信号通路相关蛋白的表达。结果与对照组比较,模型组小鼠鼓室积脓,鼓室黏膜增厚,有明显的炎症细胞浸润,中耳灌洗液中IL-6、TNF-α水平显著升高(P<0.01),AQP5蛋白表达显著降低(P<0.01),TLR4、MyD88、p-IκBα、p-p65蛋白表达显著升高(P<0.01)。与模型组比较,桉柠蒎肠溶软胶囊各剂量可以减轻鼓室积脓,减少炎症细胞浸润,明显降低中耳灌洗液中IL-6、TNF-α水平(P<0.01),显著升高AQP5蛋白表达(P<0.01),明显降低TLR4、MyD88、p-IκBα、p-p65蛋白表达(P<0.01)。结论桉柠蒎肠溶软胶囊通过提高AQP5蛋白的表达,抑制TLR4/MyD88/NF-κB信号通路的激活,减轻LPS诱导的OME。
Objective To investigate the therapeutic effect and mechanism of Eucalyptol,Limonene and Pinene Enteric Soft Capsules on otitis media with effusion(OME)model in mice based on aquaporin 5(AQP5)-Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear transcription factorκB(NF-κB)pathway.Method Kunming mice were randomly divided into control group,model group,dexamethasone group,low-,medium-and high-dose(55,110,and 220 mg·kg^(-1))groups of Eucalyptol,Limonene and Pinene Enteric Soft Capsules.The mice OME model was established by injecting LPS through the tympanic membrane.The structural changes of tympanic membrane in each group were observed microscopically.Hematoxylineosin(HE)staining was used to assess the histopathological changes of the middle ear.ELISA was used to detect the levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the middle ear lavage fluid.Western blotting was used to detect the expression of AQP5 and TLR4/MyD88/NF-κB signaling pathway-related proteins in the middle ear mucosa.Results Compared with the control group,the tympanic empyema,tympanic mucosa thickening,inflammatory cell infiltration,IL-6 and TNF-αlevels in the middle ear lavage fluid were considerably increased(P<0.01),AQP5 protein expression was significantly decreased(P<0.01),and TLR4,MyD88,p-IκBα,and p-p65 protein expression was significantly increased(P<0.01)in the model group.Compared with the model group,each dose of Eucalyptol,Limonene and Pinene Enteric Soft Capsules could reduce tympanic empyema,reduce inflammatory cell infiltration,considerably reduce IL-6 and TNF-αlevels in middle ear lavage fluid(P<0.01),significantly increase AQP5 protein expression(P<0.01),and significantly reduce TLR4,MyD88,p-IκBα,and p-p65 protein expression(P<0.01).Conclusion Eucalyptol,Limonene and Pinene Enteric Soft Capsules can reduce LPS-induced OME by increasing the expression of AQP5 protein and inhibiting the activation of TLR4/MyD88/NF-κB signaling pathway.
作者
何碧瑶
杨俊杰
党龙姣
孙萌萌
刘佳宇
辛文妤
HE Biyao;YANG Junjie;DANG Longjiao;SUN Mengmeng;LIU Jiayu;XIN Wenyu(School of Pharmacy,Binzhou Medical University,Yantai 264003,China;School of Pharmacy,Yantai University,Yantai 264005,China)
出处
《药物评价研究》
CAS
北大核心
2024年第8期1820-1826,共7页
Drug Evaluation Research
