柴胡皂苷D调节Th1/Th2平衡对小鼠急性中耳炎的治疗作用及其机制研究  

Study on the Therapeutic Effect and Mechanism of Saikosaponin D Regulating Th1/Th2 Balance on Acute Otitis Media in Mice

作　　者：张萃 蒋正举 吴应玲 丁景菊 王胜 谷雨 何磊 ZHANG Cui;JIANG Zhengju;WU Yingling;DING Jingju;WANG Sheng;GU Yu;HE Lei(Liupanshui People's Hospital,Liupanshui Guizhou 553001,China;Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563001,China)

机构地区：[1]六盘水市人民医院,贵州六盘水553001 [2]遵义医科大学附属医院,贵州遵义563001

出　　处：《中医药导报》2023年第2期24-27,34,共5页Guiding Journal of Traditional Chinese Medicine and Pharmacy

摘　　要：目的:探究柴胡皂苷D对小鼠急性中耳炎的治疗作用,并探讨其可能机制。方法:采用听泡穿刺法建立急性中耳炎模型,以随机数字表法将30只急性中耳炎小鼠分为模型组、柴胡皂苷D组、联合组,每组各10只;另取10只小鼠设为假手术组。联合组小鼠腹腔注射柴胡皂苷D(2 mg/kg),尾静脉注射脂多糖[Toll样受体4(TLR4)/髓样分化因子88(MyD88)通路激活剂](0.58 mg/kg);柴胡皂苷D组小鼠腹腔注射柴胡皂苷D(2 mg/kg),尾静脉注射等体积生理盐水;假手术组、模型组腹腔注射等体积生理盐水,尾静脉注射等体积生理盐水。1次/d,持续3 d。检测中耳灌洗液(MELF)炎症细胞计数、肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)水平;流式细胞术检测外周血Th1、Th2构成比;Western bloting法检测中耳组织IL-33、TLR4、MyD88蛋白相对表达量。结果:柴胡皂苷D组小鼠MELF炎症细胞计数,TNF-α、IL-1β水平,外周血Th2构成比,以及中耳组织IL-33、TLR4、MyD88蛋白相对表达量均低于模型组,外周血Th1构成比及Th1/Th2均高于模型组,差异均有统计学意义(P<0.05);联合组小鼠MELF炎症细胞计数,TNF-α、IL-1β水平,外周血Th2构成比,以及中耳组织IL-33、TLR4、MyD88蛋白相对表达量均高于柴胡皂苷D组,外周血Th1构成比及Th1/Th2均低于柴胡皂苷D组,差异均有统计学意义(P<0.05)。结论:柴胡皂苷D可抑制炎症反应,调节Th1/Th2平衡,从而对急性中耳炎小鼠产生治疗作用。其作用机制可能与抑制TLR4/MyD88信号通路活性有关。Objective:To explore the therapeutic effect of saikosaponin D on acute otitis media in mice,and to explore the possible mechanism.Methods:The model of acute otitis media was established by acoustic vesicle puncture.30 mice with acute otitis media were randomly divided into model group,saikosaponin D group and combination group,10 in each group.Another 10 mice were set as sham operation group.The combination group was given intraperitoneal injection of saikosaponin D(2 mg/kg),and tail vein injection of lipopolysaccharide[Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)pathway activator](0.58 mg/kg).The saikosaponin D group was intraperitoneally injected saikosaponin D(2 mg/kg),and the same volume of normal saline was injected into the tail vein.The sham operation group and the model group were injected with the same volume of normal saline intraperitoneally,and the same volume of normal saline was injected into the tail vein.Once a day for 3 days.The number of inflammatory cells,the levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in middle ear lavage fluid(MELF)were detected.Flow cytometry was used to detect the composition ratio of Th1 and Th2 in peripheral blood.Western bloting was used to detect the protein expressions of IL-33,TLR4 and MyD88 in the middle ear tissue.Results:The number of MELF inflammatory cells,the level of TNF-αand IL-1β,the peripheral blood Th2 composition ratio,and the relative expressions of IL-33,TLR4,MyD88 protein in middle ear tissue in saikosaponin D group were lower than those in the model group,the peripheral blood Th1 composition ratio and Th1/Th2 were higher than that in the model group,with statistical significance(P<0.05).The number of MELF inflammatory cells,the level of TNF-αand IL-1β,the composition ratio of peripheral blood Th2,and the relative expression of IL-33,TLR4 and MyD88 protein in the middle ear tissue of mice in the combination group were higher than those in the saikosaponin D group.The composition ratio of peripheral blo

关 键 词：急性中耳炎 柴胡皂苷D 脂多糖 TH1/TH2 Toll样受体4 髓样分化因子88 小鼠 

分 类 号：R285.5[医药卫生—中药学]