基于TLR4/MyD88/NF-κB通路研究愈疡胶囊抗中耳炎的作用  

Study on the effects of Yuyang Capsules on otitis media in rats based on TLR4/MyD88/NF-κB pathway

作　　者：罗伦才 童妍 张兴国 陈礴 钟振东 LUO Luncai;TONG Yan;ZHANG Xingguo;CHEN Bo;ZHONG Zhendong(The Second People′s Hospital of Liangshan Yi Autonomous Prefecture,Xichang 615000,China;Department of Pharmacy,School of Life Science and Engineering,Southwest Jiaotong University,Chengdu 610031,China;Institute of Laboratory Animals,Sichuan Academy of Medical Science,Sichuan Provincial People′s Hospital,Chengdu 610000,China)

机构地区：[1]凉山彝族自治州第二人民医院,西昌615000 [2]西南交通大学生命科学与工程学院药学系,成都610031 [3]四川省医学科学院,四川省人民医院实验动物研究所,成都610000

出　　处：《西北药学杂志》2021年第1期66-71,共6页Northwest Pharmaceutical Journal

摘　　要：目的构建肺炎链球菌诱导大鼠中耳炎(OM)模型,探讨愈疡胶囊延缓OM炎的作用与机制。方法体外抑菌实验测定愈疡胶囊对肺炎链球菌的最低抑菌质量浓度(MIC)与最低杀菌质量浓度(MBC)。将50只大鼠均分为模型组、阳性对照组(左氧氟沙星20 mg·kg^-1)以及愈疡胶囊高、中、低剂量组(600,300,150 mg·kg^-1),每组10只。采用鼓室内注射菌液构建模型,灌胃给药,每日1次(20 mL·kg^-1)。第7天和第14天观察鼓膜情况并收集眼眶静脉血,使用Elisa法检测血清中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、免疫球蛋白A(IgA)和免疫球蛋白G(IgG)的含量。第14天后处死大鼠,采集中耳鼓室,使用HE染色与扫描电镜观察鼓室黏膜,用Western Blot法检测鼓室Toll-样受体4(TLR4)、髓样分化蛋白-88(MyD88)和磷酸化核因子-κB(p-NF-κB)的含量。结果体外MIC与MBC分别为12.5,25.0 g·L^-1;与模型组相比,愈疡胶囊高剂量组大鼠血清中IL-1β、IL-6、TNF-α、IgA和IgG含量均降低,TLR4、MyD88与p-NF-κB蛋白含量均降低(P<0.05),鼓膜观察、HE染色与电镜观察均显示细菌感染情况减轻。结论愈疡胶囊对大鼠细菌感染OM具有抑菌抗炎作用,主要表现为减少炎症与免疫反应,其机制可能与抑制TLR4/MyD88/NF-κB信号通路相关。Objective To establish a model of otitis media(OM)in rats induced by Streptococcus pneumoniae,and to investigate the effect and mechanism of Yuyang Capsules in delaying OM.Methods The minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of Yuyang Capsules against Streptococcus pneumoniae in vitro were determined by bacteriostatic assay.50 rats were divided into model group,positive control group(levofloxacin,20 mg·kg^-1),Yuyang Capsules high,middle and low dose groups(600,300 and 150 mg·kg^-1),each group 10 rats.The model was constructed by injecting bacterial solution into the tympanic cavity,and administered by intragastric administration once a day(20 mL·kg^-1).On the 7th and 14th day,the tympanic membrane was observed and orbital venous blood was collected.The contents of nterleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),immunoglobulin A(IgA)and immunoglobulin G(IgG)in serum were detected by Elisa.The rats were sacrificed on the 14th day,and the tympanum was collected.The tympanum mucosa was observed by HE staining and scanning electron microscopy.The protein contents of toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88)and phosphorylates-nuclear factor-κB(p-NF-κB)in the tympanum were determined by Western Blot.Results The MIC and MBC were 12.5 and 25.0 g·L^-1,respectively.The contents of IL-1β,IL-6,TNF-α,IgA,IgG and protein contents of TLR4,MyD88,p-NF-κB in the Yuyang Capsules high dose group were lower than those in the model group(P<0.05).The observation of tympanic membrane,HE staining and electron microscopy all showed that the bacterial infection was alleviated.Conclusion Yuyang Capsules has bacteriostatic and anti-inflammatory effects on OM with bacterial infection in rats,which is mainly manifested by reducing inflammation and immune response.The mechanism may be related to the inhibition of TLR4/MyD88/NF-κB signaling pathway.

关 键 词：愈疡胶囊 中耳炎(OM) 抗炎 抗菌 TLR4/MyD88/NF-κB 

分 类 号：R965[医药卫生—药理学]