本文選題：醛固酮 + Af9�。� 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:梅尼埃病(MD)的典型病理特征是內(nèi)淋巴積水(EH)。醛固酮能誘導(dǎo)EH形成,但其與MD的關(guān)系尚未闡明。因此,本實(shí)驗(yàn)通過檢測(cè)豚鼠耳蝸組織中Af9蛋白的表達(dá)情況,并且分析醛固酮對(duì)耳蝸功能及Af9表達(dá)的調(diào)控作用,從而探討醛固酮與Af9分子在內(nèi)淋巴積水中的作用機(jī)制。方法:將30只豚鼠隨機(jī)分為兩組,對(duì)照組腹腔注射0.05ml生理鹽水,實(shí)驗(yàn)組腹腔注射醛固酮0.1mg.kg-1.d-1(0.05ml),各5天。1月后通過聽性腦干反應(yīng)(ABR)檢測(cè)內(nèi)耳的功能學(xué)改變;應(yīng)用蘇木素-伊紅染色(HE)檢測(cè)兩組內(nèi)耳組織的形態(tài)學(xué)改變,利用免疫組織化學(xué)(IHC)及免疫熒光(IF)檢測(cè)Af9蛋白在耳蝸組織中的定位表達(dá),采用Western blot和熒光定量PCR方法從蛋白和mRNA水平上檢測(cè)醛固酮作用前后Af9分子表達(dá)水平的變化。結(jié)果:醛固酮作用后,ABR提示實(shí)驗(yàn)組(23.33±2.38 d B SPL)聽力閾值較對(duì)照組(18.33±2.73 dB SPL)升高(P0.05)。HE證實(shí)實(shí)驗(yàn)組豚鼠耳蝸內(nèi)淋巴積水率達(dá)到73%,而對(duì)照組未見內(nèi)淋巴積水形成。IHC及IF顯示Af9蛋白在血管紋、前庭膜、螺旋緣、Corti器、螺旋神經(jīng)節(jié)均有表達(dá),并且Western blot顯示實(shí)驗(yàn)組耳蝸Af9蛋白表達(dá)量(0.75±0.15)低于對(duì)照組(1.06±0.08);熒光定量PCR顯示實(shí)驗(yàn)組耳蝸Af9mRNA(0.36±0.07)表達(dá)量低于對(duì)照組(1.00±0.00),差異均有統(tǒng)計(jì)學(xué)意義(P0.05),表明醛固酮在蛋白及mRNA水平上均能有效下調(diào)Af9分子的表達(dá)。結(jié)論:耳蝸組織中存在與Na+代謝相關(guān)的Af9分子,醛固酮對(duì)內(nèi)淋巴的調(diào)控可能與耳蝸Af9分子密切相關(guān),即醛固酮通過下調(diào)Af9的表達(dá)來參與內(nèi)淋巴代謝,形成內(nèi)淋巴積水,進(jìn)而影響耳蝸功能。
[Abstract]:Objective: the typical pathological feature of MDD is endolymphatic hydronephrosis. Aldosterone can induce the formation of EH, but the relationship between aldosterone and MD has not been clarified. Therefore, by detecting the expression of Af9 protein in guinea pig cochlea, and analyzing the effect of aldosterone on cochlea function and Af9 expression, we investigated the mechanism of aldosterone and Af9 molecule in hydrolysate. Methods: thirty guinea pigs were randomly divided into two groups: the control group was injected with 0.05ml saline intraperitoneally, and the experimental group was injected with aldosterone 0.1 mg 路kg ~ (-1) 路d ~ (-1) 路d ~ (0.05) ml for 5 days. The functional changes of the inner ear were detected by auditory brainstem response (ABR) after one month. The morphological changes of inner ear tissues were detected by hematoxylin and eosin staining. The localization expression of Af9 protein in cochlea was detected by immunohistochemistry and immunofluorescence. Western blot and fluorescence quantitative PCR were used to detect the changes of Af9 expression before and after aldosterone treatment at the level of protein and mRNA. Results: after aldosterone treatment, ABR suggested that the hearing threshold of the experimental group (23.33 鹵2.38 d B SPL) was higher than that of the control group (18.33 鹵2.73 dB SPLL). It was confirmed that the rate of endolymphatic hydrops in the guinea pigs in the experimental group was 73%, while in the control group, there was no formation of the endolymphatic hydrops and if protein was in the stria vascularis. The vestibular membrane, corti organ and spiral ganglion were all expressed. Western blot showed that the expression of Af9 protein in cochlea of experimental group was 0.75 鹵0.15) lower than that of control group (1.06 鹵0.08), and the expression of Af9mRNA(0.36 鹵0.07 in cochlea of experimental group was lower than that of control group (1.00 鹵0.007). The difference was statistically significant (P 0.05), which indicated that aldosterone was both protein and mRNA level. Can effectively down-regulate the expression of Af9 molecules. Conclusion: there are Af9 molecules related to Na metabolism in cochlea. The regulation of Aldosterone on endolymph may be closely related to Af9 molecule in cochlea, that is, aldosterone participates in endolymphatic metabolism and forms endolymphatic hydrops by down-regulating the expression of Af9. And then affect the cochlear function.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R764.33

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本文編號(hào)：1895562