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  本文選題：D半乳糖 + 衰老��； 參考：《武漢大學》2012年博士論文

【摘要】：目的：研究D-半乳糖致衰老豚鼠和自然老年豚鼠喉部細胞外基質(zhì)成分和水通道蛋白AQP1-5的表達情況,探討D-半乳糖致衰老豚鼠用于老年嗓音研究的可行性及水通道蛋白在豚鼠喉部老化過程中的作用。 方法：選取耳廓反射靈敏、無噪聲暴露史和耳毒性藥物應用史的4月齡豚鼠32只,體重300～550g,隨機分為對照組和D-半乳糖致衰老模型組,每組16只。另選相同條件自然衰老的2年齡豚鼠16只,體重600～900g,作為老年組。對照組和老年組動物不作任何處理,模型組動物頸背部皮下注射D-半乳糖300mg/kg.d (40mg/ml),每天一次,連續(xù)6周,致其衰老。然后各組動物斷頭處死,快速取出喉部,分別應用Alcian染色法、Masson三色染色法及免疫組織化學染色法觀察三組豚鼠喉部透明質(zhì)酸、膠原蛋白及水通道蛋白1～5的表達情況,并用Image-Pro Plus6.0For Window (IPP6.0)圖像分析軟件進行分析。 結(jié)果：對照組、老年組、模型組透明質(zhì)酸表達的灰度值分別為0.798351、0.694163和0.724869,三組豚鼠喉部透明質(zhì)酸的含量依次為對照組模型組老年組；膠原蛋白表達的灰度值分別為0.632295、0.71147和0.768211,三組豚鼠喉部膠原蛋白的含量依次為對照組老年組模型組,三組間差異均有統(tǒng)計學意義(P0.05)。豚鼠喉部有AQP1～5表達,AQP1主要表達在喉粘膜假復層纖毛柱狀上皮、固有層的腺體(包括漿液性和粘液性腺體)以及小血管內(nèi)皮細胞,三組表達強度依次為對照組模型組老年組,老年組未見表達；AQP2主要表達于黏膜固有層的粘液性腺體和炎性浸潤細胞,黏膜上皮細胞無或弱表達,AQP3高表達于黏膜假復層纖毛柱狀上皮,腺體、血管內(nèi)皮細胞和肌纖維組織均無表達,三組間AQP2和AQP3的表達無明顯差異；AQP4主要表達于喉粘膜鱗狀上皮、假復層柱狀纖毛上皮、固有層粘膜腺體、肌纖維及部分浸潤細胞,血管內(nèi)皮細胞無表達,三組表達強度依次為對照組模型組老年組,其中老年組基本無表達或弱表達；AQP5主要表達于喉粘膜假復層纖毛柱狀上皮細胞、上皮基底細胞、黏膜下漿液性腺體及導管,粘液性腺體無表達,三組表達強度依次為對照組老年組模型組。 結(jié)論：與4月齡豚鼠相比較,D-半乳糖所致衰老豚鼠喉部透明質(zhì)酸含量有一定程度的降低,膠原蛋白含量有一定程度的增加,與自然衰老豚鼠喉部變化一致,D-半乳糖所致衰老豚鼠可能可以用于老年嗓音的實驗研究。豚鼠喉部有AQP1～5表達,老年豚鼠和D-半乳糖致衰老模型豚鼠喉部AQP1、AQP4的表達明顯低于4月齡豚鼠,AQP5的表達明顯高于4月齡豚鼠,AQP1、AQP4和AQP5可能與豚鼠喉部組織的老化改變有關(guān)。
[Abstract]:Objective : To study the expression of AQP1 - 5 in the extracellular matrix and AQP1 - 5 in guinea pig and naturally aged guinea pig , and to investigate the feasibility of D - galactose - induced aging guinea pig for the elderly voice study and the role of water channel protein in the aging of guinea pig ' s throat .

Methods : Thirty - four months old guinea pigs were randomly divided into control group and D - galactose - induced aging model group . The rats were randomly divided into two groups : control group and D - galactose - induced aging model group .

Results : The gray values of hyaluronic acid expression in the control group , the old group and the model group were 0.798351 , 0.694163 and 0.869 respectively , and the contents of hyaluronic acid in the laryngeal part of the three groups were in turn the control group model group and the old group ;
The gray values of the collagen expression were 0.632295 , 0.71147 and 0.768211 , respectively . The contents of collagen in the laryngeal part of the guinea pigs were statistically significant ( P0.05 ) . AQP1 was expressed in the laryngeal mucosa pseudostratified ciliated columnar epithelium , the lamina propria gland ( including serous fluid and mucinous gland ) and the small vascular endothelial cells .
AQP2 was mainly expressed in mucous glands of lamina propria and inflammatory infiltrate cells , and there was no or weak expression of AQP3 , AQP3 was expressed in the ciliated columnar epithelium , glandular , vascular endothelial cells and muscle fiber tissues of the mucosa pseudostratified ciliaris , and there was no significant difference between AQP2 and AQP3 in the three groups .
AQP4 was mainly expressed in laryngeal mucosa squamous epithelium , pseudostratified columnar ciliated epithelium , lamina propria muscularis propria , muscle fibers and partially infiltrated cells , and vascular endothelial cells were not expressed .
AQP5 was mainly expressed in recurrent laryngeal mucosa pseudostratified ciliated columnar epithelial cells , epithelial basement cells , submucous serous glands and catheters , and mucinous glands were not expressed .

Conclusion : Compared with 4 - month - old guinea pigs , the content of hyaluronic acid in the laryngeal part of guinea pig due to D - galactose is reduced to some extent , and the content of collagen is increased , and the aging guinea pig caused by D - galactose can be used in the experimental study of senile voice . AQP1 - 5 expression in the laryngeal part of guinea pig , AQP1 and AQP4 expression in guinea pig and D - galactose induced aging model are obviously lower than that of 4 - month - old guinea pigs , AQP1 , AQP4 and AQP5 may be related to the aging change of the laryngeal tissue of guinea pig .

【學位授予單位】：武漢大學
【學位級別】：博士
【學位授予年份】：2012
【分類號】：R-332

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