老年性聋大鼠耳蜗中AQP1，AQP4的表达

发布时间：2018-06-26 19:12

本文选题：老年性聋 + 耳蜗　；参考：《郑州大学》2010年硕士论文

【摘要】： 目的老年性聋的患者日益增多,严重影响了老年人的生活质量,但老年性聋的发病机制尚不完全清楚,而水通道蛋白(Aquaporin, AQP)作为水通道主要作用在于参与水分子的跨膜转运,且已被证实在哺乳动物内耳有多种亚型的分布,而且内淋巴容积及内环境的稳定对正常的听力及平衡功能的维持是至关重要的,所以推测AQP对听力功能可能起着重要的作用。本实验通过检测水通道蛋白1 (AQP1)及水通道蛋白4 (AQP4)在老年性聋大鼠耳蜗中的表达情况,以期探讨老年性聋可能的发病机制,为今后研究老年性聋的治疗方法提供科学依据。材料和方法将30只成年、健康的Wistar大鼠随机分为两组：实验组和对照组,每组15只。称取每只动物的体重,实验组按照300mg/Kg的计量标准,腹腔注射D-半乳糖溶液(40mg/ml),每天2次,连续4周,对照组按同样的方法每天注射等量的生理盐水。用药前后各组动物都分别进行听性脑干反应(ABR)测试。大鼠腹腔内注射10%水合氯醛,剂量为0.1ml/300g,麻醉成功后,用生理盐水及4%多聚甲醛PBS液依次全身灌注,然后迅速断头,取出听泡并去除大部分多余骨质,在解剖过程中发现有中耳炎病史的大鼠的耳蜗标本弃用,然后将耳蜗标本置于10%EDTA液中脱钙10天,每天更换脱钙液直至耳蜗标本脱钙完全,修剪标本后用石蜡包埋,连续切片,用AQP抗体行免疫组化染色。计算机图像处理系统进行分析,SPSS统计软件进行统计学分析。结果实验组于用药后ABR反应阈值明显提高,潜伏期延长。各组动物耳蜗中AQP4于螺旋神经节细胞及Cortis'器Hensen细胞和Claudius细胞中均有分布,且实验组免疫组化阳性反应物的IOD值(积分光密度)比对照组降低,差异有统计学意义(仄O.05)。AQPt则在血管纹处呈阳性反应,但实验组IOD值较之对照组无明显改变(p0.05)。结论 AQP1、AQP4在大鼠耳蜗中有明确的分布,AQPl在血管纹处呈阳性反应,而AQP4则存在于螺旋神经节细胞及Cortis'器的Hensen细胞和Claudius细胞中。在老年性聋大鼠的耳蜗中,AQP4呈下调的趋势,而AQPl无明显变化。AQP亚型在耳蜗表达水平的变化可能改变耳蜗中的水平衡及内环境的稳定,从而影响其听力功能。
[Abstract]:Objective the increasing number of patients with presbycusis has seriously affected the quality of life of the elderly, but the pathogenesis of presbycusis is not completely clear. Aquaporin (AQP), as a water channel, is mainly involved in the transmembrane transport of water molecules, and it has been proved that there are many subtypes in mammalian inner ear. Moreover, the stability of endolymphatic volume and endolymphatic environment is very important for the maintenance of normal hearing and balance function, so we speculate that AQP may play an important role in hearing function. In this study, the expression of aquaporin-1 (AQP1) and aquaporin-4 (AQP4) in cochlea of presbycusis rats was detected in order to explore the possible pathogenesis of presbycusis and provide scientific basis for the treatment of presbycusis. Materials and methods Thirty adult and healthy Wistar rats were randomly divided into two groups: experimental group and control group with 15 rats in each group. The body weight of each animal was measured according to 300mg / kg, the experimental group received intraperitoneal injection of D-galactose solution (40mg/ml) twice a day for 4 weeks, and the control group received the same amount of normal saline daily according to the same method. Auditory brainstem response (ABR) was measured before and after treatment. Rats were injected with 10% chloral hydrate at a dose of 0.1 ml / 300 g. After anesthesia, the rats were perfused with normal saline and 4% paraformaldehyde PBS solution in turn, then the head was cut off quickly, the auditory bubble was removed and most of the excess bone was removed. The cochlear specimens of rats with history of otitis media were found to have been discarded during the anatomic process. The cochlea specimens were then decalcified in 10TA solution for 10 days. The decalcification solution was replaced every day until the cochlear specimens were decalcified completely, and the specimens were trimmed with paraffin wax. Serial sections were stained with AQP antibody. The computer image processing system was analyzed by SPSS statistical software. Results the ABR response threshold was significantly increased and the latency was prolonged in the experimental group. AQP4 was distributed in spiral ganglion cells, Hensen cells and Claudius cells in the cochlea of each group, and the IOD (integral optical density) of the immunocytochemical positive compounds in the experimental group was lower than that in the control group. The difference was statistically significant (P < 0.05). AQPt was positive in stria vascularis, but the IOD in the experimental group was not significantly different from that in the control group (p0.05). Conclusion AQP4 is positively distributed in rat cochlea, and AQP4 is present in Hensen cells and Claudius cells in spiral ganglion cells and Cortis' organ. In the cochlea of presbycusis rats, AQP4 was down-regulated, while AQPl had no obvious change. The change of AQP subtype in cochlea might change the water balance in cochlea and the stability of internal environment, thus affecting its hearing function.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R764

【参考文献】