目的 探究牡荊素對分泌性中耳炎大鼠的影響及調控機制。方法 將50只雄性SD大鼠，隨機分為對照組、模型組、牡荊素（3、12 mg/kg）組、牡荊素高劑量＋AMPK抑制劑（Compound C）組，每組各10只。除對照組外，其余各組大鼠均采取內毒素法復制分泌性中耳炎大鼠模型。造模成功后，牡荊素3、12 mg/kg組分別ip相應劑量的牡荊素；牡荊素＋Compound C組ip 12 mg/kg的牡荊素后，立即ip 20 mg/kg Compound C；對照組和模型組分別ip等量生理鹽水。連續(xù)給藥21 d后，蘇木素–伊紅（HE）染色觀察中耳黏膜組織病理學改變；酶聯(lián)免疫吸附法（ELISA）測定血清腫瘤壞死因子-α（TNF-α）、白細胞介素-6（IL-6）、白細胞介素-1β（IL-1β）、白細胞介素-18（IL-18）水平；聽覺腦干誘發(fā)電位儀測定大鼠聽力功能；Western blotting檢測中耳黏膜組織腺苷酸活化蛋白激酶/NOD樣受體蛋白3（AMPK/NLRP3）通路及細胞焦亡相關蛋白表達。結果 與模型組相比，牡荊素3、12 mg/kg組大鼠中耳黏膜組織病理損傷減輕，中耳黏膜厚度、聽力反應閾值、血清TNF-α、IL-6、IL-1β、IL-18水平均降低（P＜0.05），細胞焦亡相關蛋白[Caspase-1 p20、焦亡執(zhí)行蛋白消皮素D-N（GSDMD-N）、IL-1β、IL-18]及NLRP3蛋白表達下調，p-AMPK蛋白表達上調（P＜0.05），且呈劑量相關性。與牡荊素12 mg/kg組相比，Compound C能夠逆轉牡荊素對上述指標的改變。結論 牡荊素能夠抑制分泌性中耳炎大鼠炎癥反應，減輕中耳黏膜病理性損傷，其機制與抑制AMPK/NLRP3通路介導的細胞焦亡相關。

Objective To investigate the effect and regulatory mechanism of vitexin on otitis media with effusion rats. Methods Fifty male SD rats were randomly divided into control group, model group, vitexin (3 and 12 mg/kg) groups, and vitexin + AMPK inhibitor group (Compound C), with 10 rats in each group. Except for the control group, all other groups of rats were used to replicate a rat model of otitis media with effusion using the endotoxin method. After successful modeling, vitexin (3 and 12 mg/kg) groups of vitexin were injected intraperitoneally with different doses of 3 and 12 mg/kg of vitexin. After intraperitoneal injection 12 mg/kg of vitexin, immediately injection 20 mg/kg of Compound C into the vitexin + Compound C group. The control group and model group were injected with equal amounts of physiological saline intraperitoneally. After continuous administration for 21 days, hematoxylin eosin (HE) staining was used to observe pathological changes in the middle ear mucosa tissue, Enzyme linked immunosorbent assay (ELISA) for the determination of serum TNF-α, IL-6, IL-1β, and IL-18 levels. The auditory brainstem response instrument was used to measure the hearing function of rats, Western blotting was used to detect the expression of AMPK/NLRP3 pathway and pyroptosis related proteins in the middle ear mucosa tissue. Results Compared with the model group, the pathological damage of the middle ear mucosa tissue was reduced in the vitexin (3 and 12 mg/kg) groups, and the thickness of the middle ear mucosa, hearing response threshold, and serum TNF-α, IL-6, IL-6, IL-18 levels were decreased (P < 0.05), and pyroptosis related proteins (caspase-1 p20, GSDMD-N,IL-1β, IL-18) and NLRP3 protein expression were downregulated, while p-AMPK protein expression was upregulated (P<0.05), and there was a dose-dependent relationship. Compared with the vitexin 12 mg/kg group, Compound C can reverse the changes in the above indicators caused by vitexin. Conclusion Vitexin can inhibit the inflammatory response and alleviate pathological damage to the middle ear mucosa in rats with otitis media with effusion, and its mechanism is related to the inhibition of AMPK/NLRP3 pathway mediated pyroptosis.

R285

河南省高等學校重點科研項目(20B320117)