醛固酮对豚鼠耳蜗功能及Af9表达的调控作用

发布时间：2018-05-16 05:05

  本文选题：醛固酮 + Af9　； 参考：《重庆医科大学》2017年硕士论文

【摘要】：目的:梅尼埃病(MD)的典型病理特征是内淋巴积水(EH)。醛固酮能诱导EH形成,但其与MD的关系尚未阐明。因此,本实验通过检测豚鼠耳蜗组织中Af9蛋白的表达情况,并且分析醛固酮对耳蜗功能及Af9表达的调控作用,从而探讨醛固酮与Af9分子在内淋巴积水中的作用机制。方法:将30只豚鼠随机分为两组,对照组腹腔注射0.05ml生理盐水,实验组腹腔注射醛固酮0.1mg.kg-1.d-1(0.05ml),各5天。1月后通过听性脑干反应(ABR)检测内耳的功能学改变;应用苏木素-伊红染色(HE)检测两组内耳组织的形态学改变,利用免疫组织化学(IHC)及免疫荧光(IF)检测Af9蛋白在耳蜗组织中的定位表达,采用Western blot和荧光定量PCR方法从蛋白和mRNA水平上检测醛固酮作用前后Af9分子表达水平的变化。结果:醛固酮作用后,ABR提示实验组(23.33±2.38 d B SPL)听力阈值较对照组(18.33±2.73 dB SPL)升高(P0.05)。HE证实实验组豚鼠耳蜗内淋巴积水率达到73%,而对照组未见内淋巴积水形成。IHC及IF显示Af9蛋白在血管纹、前庭膜、螺旋缘、Corti器、螺旋神经节均有表达,并且Western blot显示实验组耳蜗Af9蛋白表达量(0.75±0.15)低于对照组(1.06±0.08);荧光定量PCR显示实验组耳蜗Af9mRNA(0.36±0.07)表达量低于对照组(1.00±0.00),差异均有统计学意义(P0.05),表明醛固酮在蛋白及mRNA水平上均能有效下调Af9分子的表达。结论:耳蜗组织中存在与Na+代谢相关的Af9分子,醛固酮对内淋巴的调控可能与耳蜗Af9分子密切相关,即醛固酮通过下调Af9的表达来参与内淋巴代谢,形成内淋巴积水,进而影响耳蜗功能。
[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 卤2.38 d B SPL) was higher than that of the control group (18.33 卤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 卤0.15) lower than that of control group (1.06 卤0.08), and the expression of Af9mRNA(0.36 卤0.07 in cochlea of experimental group was lower than that of control group (1.00 卤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.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R764.33

【参考文献】

相关期刊论文 前4条

1 蒋子栋;高帆;孙程成;;不同方法构建豚鼠膜迷路积水模型的对比研究[J];中华耳科学杂志;2015年04期

2 Wenzheng Zhang;;Epigenetics of epithelial Na~+ channel-dependent sodium uptake and blood pressure regulation[J];World Journal of Nephrology;2015年03期

3 温雅;钟时勋;;去氨加压素对耳蜗功能及上皮钠通道表达的调节作用[J];中华耳科学杂志;2014年03期

4 蒋子栋,张连山;醛固酮诱发豚鼠双耳膜迷路积水[J];中国医学科学院学报;2002年05期

，

本文编号：1895562