仙芪眩宁颗粒对豚鼠耳蜗膜迷路积水AQP2表达的影响

Effects of Xianqi Xuanning Granule on the expression of aquaporin-2 in the cochlea of guinea pig with endolymphatic hydrops

关键词:  仙芪眩宁颗粒  膜迷路积水  水通道蛋白2

Key words:  Xianqi Xuanning Granule  endolymphatic hydrops  aquaporin-2

摘要:

[目的] 观察仙芪眩宁颗粒对耳蜗膜迷路积水豚鼠模型水通道蛋白2（AQP2）表达的影响并探讨其治疗梅尼尔病的作用机制。[方法] 50只健康豚鼠随机平均分为5组：正常对照组、模型组、低、中、高剂量治疗组，除对照组外均采取腹腔注射醋酸去氨加压素构建膜迷路积水豚鼠模型，3个治疗组按豚鼠体质量胃饲，药物剂量分别为6.1、12.2、24.4 g/kg。听性脑干反应（ABR）检测豚鼠听功能；膜迷路积水程度通过耳蜗切片苏木精-伊红（HE）染色，计算中阶面积与中阶和前庭阶面积和的比值R；采用免疫组织化学和蛋白印迹（Western Blot）法检测豚鼠耳蜗AQP2表达水平。[结果] 模型组听力受损ABR阈值升高，各治疗组ABR阈值与对照组相近（P>0.05）；耳蜗切片模型组R值高于对照组（P<0.01），而各治疗组R值与对照组相比未见统计学差异（P>0.05）；扫描电镜结果显示模型组出现毛细胞损伤，治疗组趋于正常；免疫组织化学法显示模型组内耳AQP2表达水平高于对照组和各治疗组（P<0.01）；Western blot结果显示模型组、低剂量治疗组AQP2表达高于对照组，而其在高剂量治疗组的表达低于对照组（P<0.01）。[结论] 仙芪眩宁颗粒可减轻豚鼠膜迷路积水程度，其机制可能与AQP2表达的下调有关。

Abstract:

[Objective] To study the influence of Xianqi Xuanning Granule on the morphology of cochlea and the expression of aquaporin-2 (AQP2) in guinea pigs with endolymphatic hydrops, and to discuss the mechanism of treating meniere's disease.[Methods] Fifty guinea pigs were randomly divided into the control group, model group, low dose treatment group(LT), moderate dose treatment group (MT), high dose treatment group (HT) on average. All guinea pigs except those in the control group were used for making endolymphatic hydrops models with intraperitoneal injection of desmopressin acetate. Three treatment groups were treated with different dosage of Xianqi Xuanning Granule by feeding way, separately 6.1, 12.2, 24.4 g/kg, while those in the control and model groups were treated with physiological saline. Auditory function was tested by auditory brain response (ABR). The endolymphatic hydrops in cochlea were evaluated by hematoxylin-eosin (HE) staining, and the ratio of the scala media area to the total area of that and scala vestibuli was calculated. The expression of AQP2 protein in guinea pig cochlea was detected by immunohistochemistry and Western Blot.[Results] The ABR threshold in the model group increased because of hearing impairment, which of each treatment group was similar to that of the control group (P>0.05). R value in the model group was higher than that in the control group (P<0.01). There was no statistical difference between the treatment groups and the control group (P>0.05). The damage of cochlea hair cells occurred in model group by scanning electron microscope, and they appeared to be normal in the treatment groups. The AQP2 expression of the model group was higher than those of the control group and three treatment groups by immunohistochemistry (P<0.01). Western Blot results showed that the expression of AQP2 in the model group and LT group was higher than that in the control group, while it in HT group was significantly lower than that in the control group (P<0.01).[Conclusion] Xianqi Xuanning Granule could suppress the development of endolymphatic hydrops in guinea pigs, and it might be associated with the down-regulation of the AQP2 expression.