本文選題：老年性聾 + 耳蝸��； 參考：《鄭州大學(xué)》2010年碩士論文

【摘要】： 目的 老年性聾的患者日益增多,嚴(yán)重影響了老年人的生活質(zhì)量,但老年性聾的發(fā)病機(jī)制尚不完全清楚,而水通道蛋白(Aquaporin, AQP)作為水通道主要作用在于參與水分子的跨膜轉(zhuǎn)運(yùn),且已被證實(shí)在哺乳動物內(nèi)耳有多種亞型的分布,而且內(nèi)淋巴容積及內(nèi)環(huán)境的穩(wěn)定對正常的聽力及平衡功能的維持是至關(guān)重要的,所以推測AQP對聽力功能可能起著重要的作用。本實(shí)驗(yàn)通過檢測水通道蛋白1 (AQP1)及水通道蛋白4 (AQP4)在老年性聾大鼠耳蝸中的表達(dá)情況,以期探討老年性聾可能的發(fā)病機(jī)制,為今后研究老年性聾的治療方法提供科學(xué)依據(jù)。 材料和方法 將30只成年、健康的Wistar大鼠隨機(jī)分為兩組：實(shí)驗(yàn)組和對照組,每組15只。稱取每只動物的體重,實(shí)驗(yàn)組按照300mg/Kg的計(jì)量標(biāo)準(zhǔn),腹腔注射D-半乳糖溶液(40mg/ml),每天2次,連續(xù)4周,對照組按同樣的方法每天注射等量的生理鹽水。用藥前后各組動物都分別進(jìn)行聽性腦干反應(yīng)(ABR)測試。大鼠腹腔內(nèi)注射10%水合氯醛,劑量為0.1ml/300g,麻醉成功后,用生理鹽水及4%多聚甲醛PBS液依次全身灌注,然后迅速斷頭,取出聽泡并去除大部分多余骨質(zhì),在解剖過程中發(fā)現(xiàn)有中耳炎病史的大鼠的耳蝸標(biāo)本棄用,然后將耳蝸標(biāo)本置于10%EDTA液中脫鈣10天,每天更換脫鈣液直至耳蝸標(biāo)本脫鈣完全,修剪標(biāo)本后用石蠟包埋,連續(xù)切片,用AQP抗體行免疫組化染色。計(jì)算機(jī)圖像處理系統(tǒng)進(jìn)行分析,SPSS統(tǒng)計(jì)軟件進(jìn)行統(tǒng)計(jì)學(xué)分析。 結(jié)果 實(shí)驗(yàn)組于用藥后ABR反應(yīng)閾值明顯提高,潛伏期延長。各組動物耳蝸中AQP4于螺旋神經(jīng)節(jié)細(xì)胞及Cortis'器Hensen細(xì)胞和Claudius細(xì)胞中均有分布,且實(shí)驗(yàn)組免疫組化陽性反應(yīng)物的IOD值(積分光密度)比對照組降低,差異有統(tǒng)計(jì)學(xué)意義(仄O.05)。AQPt則在血管紋處呈陽性反應(yīng),但實(shí)驗(yàn)組IOD值較之對照組無明顯改變(p0.05)。 結(jié)論 AQP1、AQP4在大鼠耳蝸中有明確的分布,AQPl在血管紋處呈陽性反應(yīng),而AQP4則存在于螺旋神經(jīng)節(jié)細(xì)胞及Cortis'器的Hensen細(xì)胞和Claudius細(xì)胞中。在老年性聾大鼠的耳蝸中,AQP4呈下調(diào)的趨勢,而AQPl無明顯變化。AQP亞型在耳蝸表達(dá)水平的變化可能改變耳蝸中的水平衡及內(nèi)環(huán)境的穩(wěn)定,從而影響其聽力功能。
[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.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R764

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