[關(guān)鍵詞]
[摘要]
目的 基于水通道蛋白5(AQP5)-Toll樣受體4(TLR4)/髓樣分化因子88(MyD88)/核轉(zhuǎn)錄因子κB(NF-κB)通路探討桉檸蒎腸溶軟膠囊對小鼠分泌性中耳炎(OME)模型的治療作用及機(jī)制。方法 昆明小鼠隨機(jī)分為對照組、模型組、地塞米松組和桉檸蒎腸溶軟膠囊低、中、高劑量(55、110、220 mg·kg-1)組。采用經(jīng)鼓膜注入脂多糖(LPS)的方法建立小鼠OME模型,顯微鏡觀察各組小鼠鼓膜結(jié)構(gòu)變化,蘇木精-伊紅(HE)染色法評估中耳組織病理學(xué)改變,ELISA法檢測中耳灌洗液中白細(xì)胞介素-6(IL-6)、腫瘤壞死因子-α(TNF-α)水平,Western blotting檢測中耳黏膜組織AQP5及TLR4/MyD88/NF-κB信號通路相關(guān)蛋白的表達(dá)。結(jié)果 與對照組比較,模型組小鼠鼓室積膿,鼓室黏膜增厚,有明顯的炎癥細(xì)胞浸潤,中耳灌洗液中IL-6、TNF-α水平顯著升高(P<0.01),AQP5蛋白表達(dá)顯著降低(P<0.01),TLR4、MyD88、p-IκBα、p-p65蛋白表達(dá)顯著升高(P<0.01)。與模型組比較,桉檸蒎腸溶軟膠囊各劑量可以減輕鼓室積膿,減少炎癥細(xì)胞浸潤,明顯降低中耳灌洗液中IL-6、TNF-α水平(P<0.01),顯著升高AQP5蛋白表達(dá)(P<0.01),明顯降低TLR4、MyD88、p-IκBα、p-p65蛋白表達(dá)(P<0.01)。結(jié)論 桉檸蒎腸溶軟膠囊通過提高AQP5蛋白的表達(dá),抑制TLR4/MyD88/NF-κB信號通路的激活,減輕LPS誘導(dǎo)的OME。
[Key word]
[Abstract]
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 factor κB (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-α levels 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-α levels 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.
[中圖分類號]
R285.5
[基金項(xiàng)目]