本文選題：老年性聾 + 晚期糖基化終產(chǎn)物受體　； 參考：《福建醫(yī)科大學(xué)》2012年碩士論文

【摘要】：老年性聾病因及發(fā)病機(jī)制復(fù)雜，主要是衰老、凋亡、氧化應(yīng)激和線粒體突變四大假說(shuō)，其中衰老的自由基學(xué)說(shuō)被較多人所接受。但產(chǎn)生自由基的原因甚多，分子機(jī)制尚未統(tǒng)一，因此目前臨床老年性聾的治療不容樂觀。RAGE與AGEs結(jié)合可引起細(xì)胞內(nèi)氧化應(yīng)激和轉(zhuǎn)錄因子NF-κB的激活，并可進(jìn)一步增強(qiáng)RAGE表達(dá)[15]，從而引起持續(xù)的細(xì)胞損傷和功能紊亂。RAGE介導(dǎo)的氧化應(yīng)激在中樞神經(jīng)系統(tǒng)損傷性疾病中至關(guān)重要，而在衰老的內(nèi)耳中研究較少。本研究擬通過(guò)老年性聾動(dòng)物模型，探討RAGE及其介導(dǎo)信號(hào)途徑與老年性聾發(fā)生發(fā)展的關(guān)系。主要內(nèi)容分以下三部分進(jìn)行。 第一部分：C57BL/6J小鼠年齡相關(guān)性聽功能和內(nèi)耳組織學(xué)的研究目的：研究不同月齡的C57BL/6J小鼠的聽功能和內(nèi)耳組織學(xué)特點(diǎn)，探討老年性聾發(fā)病的動(dòng)物模型。 方法：將C57BL/6J小鼠分為2月齡（幼年）、4月齡（中）、10月齡（老年）組，每組10只，分別行ABR閾值檢測(cè)聽力水平，組織學(xué)切片觀察耳蝸形態(tài)結(jié)構(gòu)，基底膜鋪片觀察耳蝸Corti器。 結(jié)果：（1）聽力學(xué)檢查示：C57BL/6J小鼠ABR反應(yīng)閾值在2月齡組為10±1.24dBnHL；4月齡組時(shí)ABR反應(yīng)閾值上升到18±1.87dBnHL，與2月齡組相比較，p0.05，有統(tǒng)計(jì)學(xué)意義；10月齡組聽力損失60±2.91dBnHL，，與4月齡組相比，p0.01，差異有顯著的統(tǒng)計(jì)學(xué)意義；(2) HE染色見螺旋神經(jīng)節(jié)細(xì)胞隨年齡增大逐漸減少，到10月齡時(shí)明顯較2、4月齡減少，內(nèi)外毛細(xì)胞不同程度丟失，血管紋在老年組的小鼠內(nèi)耳中變細(xì)窄；（3）基底膜鋪片示隨年齡增長(zhǎng)內(nèi)外毛細(xì)胞由低回逐漸向頂回有所缺失，老年鼠外毛細(xì)胞缺失先于內(nèi)毛細(xì)胞； 結(jié)論：C57BL/6J小鼠隨年齡增大呈老年性聾的聽力變化。 第二部分：C57BL/6J小鼠血清AGEs、內(nèi)耳S100B和RAGE表達(dá)的相關(guān)性研究目的：探討C57BL/6J小鼠內(nèi)耳RAGE表達(dá)和AGEs、S100B是否具有相關(guān)性。方法：將C57BL/6J小鼠分為2月齡、4月齡、10月齡組，每組10只，采用ELISA法檢測(cè)血清中AGEs含量，半定量RT-PCR檢測(cè)S100B在耳蝸中的表達(dá)，Real time PCR檢測(cè)RAGE在耳蝸中表達(dá)情況。 結(jié)果：（1）2月齡組小鼠血清AGEs濃度是71.33±7.09ng/L，4月齡組105.44±12.1ng/L，10月齡組226.67±7.63ng/L，經(jīng)兩兩比較，p0.05，差異有統(tǒng)計(jì)學(xué)意義；（2）S100B在2月齡小鼠耳蝸中的灰度值是31836±363,24.72,在10月齡小鼠耳蝸中的灰度值是67449.66±1988.61，P0.05，差異有統(tǒng)計(jì)學(xué)意義。（3）Real time PCR檢測(cè)RAGEmRNA表達(dá)結(jié)果示：將2月齡組小鼠內(nèi)耳RAGE表達(dá)量為1定為標(biāo)準(zhǔn)對(duì)照組，4月齡組小鼠內(nèi)耳RAGE相對(duì)量1.61±0.07，10月齡組RAGE相對(duì)量3.13±0.08，兩兩比較p0.05，P均有統(tǒng)計(jì)學(xué)意義。結(jié)論：C57BL/6J小鼠隨年齡增大血清AGEs含量增多，內(nèi)耳S100B表達(dá)增高，而促使它們的受體RAGE表達(dá)增多，說(shuō)明內(nèi)耳RAGE表達(dá)和配體含量具有相關(guān)性。 第三部分：RAGE介導(dǎo)的信號(hào)通路在老年性聾的作用研究 目的：通過(guò)研究RAGE、NF-κB、P21在小鼠耳蝸中的表達(dá)，探討RAGE信號(hào)途徑與老年性聾發(fā)生的關(guān)系。 方法：將C57BL/6J小鼠分為2月齡、4月齡、10月齡組，每組10只，免疫組化檢測(cè)RAGE、NF-κB、P21在耳蝸中的定位表達(dá)，采用圖像分析軟件計(jì)算所染螺旋神經(jīng)節(jié)細(xì)胞和血管紋的平均光密度值。 結(jié)果：免疫組化結(jié)果見RAGE、NF-κB、P21主要在小鼠耳蝸的螺旋神經(jīng)節(jié)細(xì)胞、Corti器和血管紋中表達(dá)；在耳蝸螺旋神經(jīng)節(jié)細(xì)胞和血管紋中10月齡組平均光密度值比4月齡組大，4月齡組平均光密度值比2月齡大，p0.05，差異有統(tǒng)計(jì)學(xué)意義。 結(jié)論：RAGE、NF-κB、P21在老化型小鼠耳蝸中均有表達(dá)且隨年齡增大表達(dá)增多，說(shuō)明RAGE、NF-κB、P21均參與老年性聾的發(fā)病過(guò)程，可能RAGE介導(dǎo)NF-κB、P21等相關(guān)蛋白導(dǎo)致聽覺細(xì)胞丟失或損傷，表明RAGE介導(dǎo)的信號(hào)途徑與老年性聾的發(fā)病過(guò)程有相關(guān)性。
[Abstract]:The etiology and pathogenesis of senile deafness are complex, mainly four hypotheses of aging, apoptosis, oxidative stress and mitochondrial mutation, of which the free radical theory of aging is accepted by many people. However, there are many reasons for the generation of free radicals and the molecular mechanism is not unified. Therefore, the treatment of clinical senile deafness can not be optimistic about the combination of.RAGE and AGEs. The activation of intracellular oxidative stress and transcription factor NF- kappa B can further enhance the RAGE expression of [15], thus causing persistent cell damage and dysfunction of.RAGE mediated oxidative stress in the central nervous system injury diseases, but less in the aging inner ear. This study is to explore RA through the senile deafness animal model. The relationship between GE and its mediating signal pathway and the occurrence and development of senile deafness is divided into three parts.
The first part: the study of age related auditory function and inner ear histology in C57BL/6J mice: To study the auditory and inner ear histology of C57BL/6J mice of different months of age and to explore the animal models of the onset of senile deafness.
Methods: the C57BL/6J mice were divided into 2 month old (young), 4 month old (middle), 10 month old (old) group and 10 in each group. The hearing level was measured by ABR threshold, and the cochlear morphology was observed by histological section. The cochlear Corti was observed by the basement membrane sheet.
Results: (1) the audiological examination showed that the threshold of ABR reaction in C57BL/6J mice was 10 + 1.24dBnHL, and the threshold of ABR reaction increased to 18 + 1.87dBnHL in the 4 month old group. Compared with the 2 month old group, P0.05 was statistically significant; the 10 month old group of hearing loss was 60 + 2.91dBnHL, and the difference was statistically significant compared with the 4 month old group, and (2) H. E staining showed that the spiral ganglion cells decreased gradually with age, at 10 month old, obviously less than 2,4 months, the internal and external hair cells were lost in different degrees, and the vascular lines were narrowed in the inner ear of the old mice. (3) the basal membrane sheets showed the loss of the inner and outer hair cells from the low back to the top, and the loss of outer hair cells in the aged rats. Prior to internal hair cells;
Conclusion: C57BL/6J mice showed hearing loss with age.
The second part: the correlation of C57BL/6J mice serum AGEs, S100B and RAGE expression in the inner ear: To investigate the expression of RAGE in the inner ear of C57BL/6J mice and the correlation between AGEs and S100B. Methods: C57BL/6J mice were divided into 2 month old, 4 month old, 10 month old, and 10 groups. The AGEs content in the serum was detected by ELISA method, and the semi quantitative RT-PCR was detected. The expression of B in the cochlea and Real time PCR were used to detect the expression of RAGE in the cochlea.
Results: (1) the concentration of AGEs in the 2 month old groups of mice was 71.33 + 7.09ng/L, 4 month old 105.44 + 12.1ng/L, 10 month old groups 226.67 + 7.63ng/L, and the difference was statistically significant after 22. (2) the gray value of S100B in the cochlea of 2 month old mice was 2 month old 363,24.72, and the gray value in the cochlea of 10 month old mice was 67449.66 + 1988.61, P0.05, The difference was statistically significant. (3) Real time PCR detection RAGEmRNA expression results showed that the 2 month old groups of mice internal ear RAGE expression of 1 was set as the standard control group, 4 month old groups of mice inner ear RAGE relative quantity 1.61 + 0.07,10 month group RAGE relative quantity 3.13 + 0.08, 22 P0.05, P all statistical significance. Conclusion: C57BL/6J mice with age increases. The serum AGEs level increased, the expression of S100B increased in the inner ear, and the expression of RAGE increased, indicating that the expression of RAGE in the inner ear was related to the ligand content.
The third part: the role of RAGE mediated signal transduction pathway in senile deafness.
Objective: To study the expression of RAGE, NF- kappa B and P21 in mouse cochlea, and to explore the relationship between RAGE signal pathway and the occurrence of senile deafness.
Methods: C57BL/6J mice were divided into 2 month old, 4 month old, 10 month old groups, each group of 10. Immunohistochemistry was used to detect RAGE, NF- kappa B, P21 in the cochlea, and the mean optical density of the infected spiral ganglion cells and vascular lines was calculated by the image analysis software.
Results: the results of immunohistochemistry were RAGE, NF- kappa B, P21 mainly in the spiral ganglion cells of the cochlea, Corti and vascular lines, and the average optical density of 10 month old groups in the cochlear spiral ganglion cells and vascular lines was larger than that of the 4 month old groups, and the average optical density of the 4 month old groups was larger than that of 2 month old, and the difference was statistically significant.
Conclusion: RAGE, NF- kappa B and P21 are expressed in the cochlea of aging mice and increase with age, indicating that RAGE, NF- kappa B, P21 are all involved in the pathogenesis of senile deafness, and RAGE mediates NF- kappa B, P21 and other related proteins lead to the loss or damage of auditory cells, indicating that the signaling pathway is related to the process of senile deafness. Sex.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R764.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 李明棟;劉兆華;;慶大霉素作用下豚鼠耳蝸螺旋神經(jīng)節(jié)細(xì)胞bcl-2及Bax的表達(dá)[J];重慶醫(yī)學(xué);2006年05期

2 樊建剛;唐s

本文編號(hào)：1954190