地骨皮甲素调节IL-33/ST2信号对中耳炎大鼠的研究

Effects of IL-33/ST2 signal on immune function and sodium channel in otitis media rats by using tiosteopantin

目的探讨地骨皮甲素调节IL-33/ST2信号对中耳炎大鼠免疫功能及钠通道的研究。方法50只SD大鼠,随机分为NO组(正常大鼠灌胃生理盐水),MO组(模型大鼠灌胃生理盐水),OS组(模型灌胃40 mg·kg^(-1)地骨皮甲素),ST组(模型灌胃0.2 mL 1 kg·L^(-1)IL-33/ST2信号抑制剂),UN组(模型灌胃40 mg·kg^(-1)地骨皮甲素和0.2 mL 1 kg·L^(-1)IL-33/ST2信号抑制剂)。听性脑干反应检测听力水平,HE染色检测中耳黏膜组织病理形态,ELISA法检测免疫功能相关指标,Real-time PCR法检测中耳黏膜组织上皮钠通道mRNA表达,免疫印迹法检测IL-33/ST2信号相关蛋白表达。结果与NO组相比,MO组纯音听力阈值、IL-4含量、IL-33、ST2蛋白表达升高(P<0.05),INF-γ含量、α-ENaC、β-ENaC、γ-ENaC mRNA表达降低(P<0.05),与MO组比较,OS组、ST组、UN组纯音听力阈值、IL-4含量、IL-33、ST2蛋白表达降低(P<0.05),INF-γ含量、α-ENaC、β-ENaC、γ-ENaC mRNA表达升高(P<0.05),且ST组与OS组相比无明显差异(P>0.05),UN组比ST组降低明显(P<0.05);与NO组相比,MO组中耳黏膜出现病理变化,与MO组相比,OS组、ST组、UN组病理症状明显改善。结论地骨皮甲素可显著改善中耳炎大鼠免疫功能,促进钠通道表达,其作用机制可能与抑制IL-33/ST2信号通路有关。

Aim To investigate the effects of IL-33/ST2 signal regulation by osteomortin on immune function and sodium channel in otitis media rats.Methods Fifty SD rats were randomly divided into NO group(normal rats with gavage of normal saline),MO group(model rats with gavage of normal saline),OS group(model rats with gavage of 40 mg·kg^(-1)desortolin),ST group(model rats with gavage of 0.2 mL 1 kg·L^(-1)IL-33/ST2 signal inhibitor),and UN group(model with administration of 40 mg·kg^(-1)desortin and 0.2 mL 1 kg·L^(-1)IL-33/ST2 signaling inhibitor).The auditory brain stem reaction was used to detect the hearing level,HE staining was used to detect the pathological morphology of middle ear mucosa,ELISA was used to detect the related indexes of immune function,Real-time PCR was used to detect the mRNA expression of sodium channel in middle ear mucosa,and Western blot was employed to detect the expression of IL-33/ST2 signal-related proteins.Results Compared with NO group,pure tone hearing threshold,IL-4 content,IL-33 and ST2 protein expression were up-regulated in MO group(P<0.05),while INF-γcontent,α-ENaC,β-ENaC andγ-ENaC mRNA expression were down-regulated in MO group(P<0.05).Compared with the MO group,pure tone hearing threshold,IL-4 content,IL-33 and ST2 protein expression in OS,ST and UN groups decreased(P<0.05),INF-γcontent,α-ENaC,β-ENaC andγ-ENaC mRNA expression increased(P<0.05),and there was no significant difference between the ST group and the OS group(P>0.05).The UN group was significantly lower than the ST group(P<0.05).Compared with NO group,the middle ear mucosa of MO group showed pathological changes,and the pathological symptoms of OS group,ST group and UN group were significantly improved compared with those of the MO group.Conclusion The immune function and sodium channel expression of otitis media rats can be significantly improved by corticosteroids,the mechanism of which may be related to the inhibition of IL-33/ST2 signal.