目的 探究牡荆素对分泌性中耳炎大鼠的影响及调控机制。方法 将50只雄性SD大鼠，随机分为对照组、模型组、牡荆素（3、12 mg/kg）组、牡荆素高剂量＋AMPK抑制剂（Compound C）组，每组各10只。除对照组外，其余各组大鼠均采取内毒素法复制分泌性中耳炎大鼠模型。造模成功后，牡荆素3、12 mg/kg组分别ip相应剂量的牡荆素；牡荆素＋Compound C组ip 12 mg/kg的牡荆素后，立即ip 20 mg/kg Compound C；对照组和模型组分别ip等量生理盐水。连续给药21 d后，苏木素–伊红（HE）染色观察中耳黏膜组织病理学改变；酶联免疫吸附法（ELISA）测定血清肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）、白细胞介素-18（IL-18）水平；听觉脑干诱发电位仪测定大鼠听力功能；Western blotting检测中耳黏膜组织腺苷酸活化蛋白激酶/NOD样受体蛋白3（AMPK/NLRP3）通路及细胞焦亡相关蛋白表达。结果 与模型组相比，牡荆素3、12 mg/kg组大鼠中耳黏膜组织病理损伤减轻，中耳黏膜厚度、听力反应阈值、血清TNF-α、IL-6、IL-1β、IL-18水平均降低（P＜0.05），细胞焦亡相关蛋白[Caspase-1 p20、焦亡执行蛋白消皮素D-N（GSDMD-N）、IL-1β、IL-18]及NLRP3蛋白表达下调，p-AMPK蛋白表达上调（P＜0.05），且呈剂量相关性。与牡荆素12 mg/kg组相比，Compound C能够逆转牡荆素对上述指标的改变。结论 牡荆素能够抑制分泌性中耳炎大鼠炎症反应，减轻中耳黏膜病理性损伤，其机制与抑制AMPK/NLRP3通路介导的细胞焦亡相关。

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

河南省高等学校重点科研项目(20B320117)