本文關(guān)鍵詞： 三叉神經(jīng) 動(dòng)物模型 神經(jīng)營養(yǎng)性角膜炎 角膜神經(jīng) 共焦顯微鏡　出處：《鄭州大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：目的 建立小鼠三叉神經(jīng)損傷的動(dòng)物模型,模擬神經(jīng)營養(yǎng)性角膜炎的發(fā)病過程和轉(zhuǎn)歸；觀察去三叉神經(jīng)小鼠角膜的臨床改變和修復(fù)過程；探討去神經(jīng)支配對(duì)小鼠角膜神經(jīng)密度、角膜上皮完整性和功能的影響。 材料與方法 選取成年雄性C57BL/6小鼠70只,按隨機(jī)數(shù)字表分為正常對(duì)照組、Id、3d、1w、2w、3w、4w,共7組,每組各10只小鼠。選取左側(cè)為實(shí)驗(yàn)側(cè),無水乙醇法去除小鼠左側(cè)三叉神經(jīng)第一支眼神經(jīng)。分別對(duì)正常對(duì)照組和不同時(shí)間點(diǎn)的實(shí)驗(yàn)組進(jìn)行淚液分泌實(shí)驗(yàn)檢查、角膜熒光素染色檢查和活體共焦顯微鏡檢查,采集照片并記錄結(jié)果。然后,過量戊巴比妥鈉腹腔注射處死小鼠,分別取小鼠左側(cè)角膜和顱內(nèi)三叉神經(jīng)眼支。小鼠角膜神經(jīng)采用Tubulinβ-Ⅲ抗體標(biāo)記,離體共聚焦顯微鏡觀察,拍攝并保存圖片；三叉神經(jīng)采用麗春紅—亮綠法染色,免疫熒光顯微鏡下觀察,拍攝并保存圖片。對(duì)所得的結(jié)果和圖片進(jìn)行統(tǒng)計(jì)和分析。 結(jié)果 1.小鼠角膜熒光素染色并裂隙燈顯微鏡下鈷藍(lán)光觀察顯示,損毀三叉神經(jīng)后小鼠角膜起初呈現(xiàn)淺層點(diǎn)狀熒光素著色,隨著病程的發(fā)展,熒光素著色灶逐漸融合,范圍擴(kuò)大,于1周時(shí)熒光素滲透性達(dá)頂峰,而后呈現(xiàn)出緩慢的恢復(fù)跡象。 2.損傷小鼠三叉神經(jīng)后,小鼠的淚液分泌量迅速減少,在2周時(shí)降為最低值,而后呈現(xiàn)出緩慢的恢復(fù)過程。 3.活體共焦顯微鏡觀察可見,損傷三叉神經(jīng)后,小鼠角膜上皮細(xì)胞體積增大,多形性明顯增加；前彈力層神經(jīng)纖維分布稀疏或不可見；角膜基質(zhì)結(jié)構(gòu)被破壞,可見其中分布大量胞體較小的高反光的細(xì)胞形物；后彈力層可以觀察到明顯的細(xì)胞皺褶結(jié)構(gòu)。 4.小鼠角膜神經(jīng)Tubulinβ-Ⅲ抗體染色顯示,損傷小鼠三叉神經(jīng)后,中央部角膜神經(jīng)密度最敏感,顯著下降；旁中央及周邊部角膜神經(jīng)也有明顯缺失,但不如中央部位表現(xiàn)顯著；病程后期未見觀察到明顯的神經(jīng)纖維的修復(fù)及再生現(xiàn)象。 5.三叉神經(jīng)冰凍切片病理染色的觀察顯示,損毀小鼠三叉神經(jīng)后,無論是橫切面還是縱切面觀察,神經(jīng)髓鞘的密度均隨著病程的進(jìn)展不同程度下降,神經(jīng)髓鞘的形態(tài)也發(fā)生顯著性改變。 結(jié)論 1.三叉神經(jīng)損傷后,小鼠角膜基底下神經(jīng)纖維叢密度顯著下降,淚膜完整性和功能受到破壞,角膜熒光素滲透性增加；小鼠角膜呈現(xiàn)神經(jīng)營養(yǎng)性角膜炎的改變。 2.隨著病程的進(jìn)展,小鼠角膜的淚膜完整性及熒光素滲透性均呈現(xiàn)出緩慢的恢復(fù)過程,而小鼠角膜基底下神經(jīng)纖維叢密度沒有觀察到明顯的增加,其可能的原因是腦源性的神經(jīng)營養(yǎng)因子和神經(jīng)生長因子分泌增加,小鼠的角膜上皮修復(fù)較快而神經(jīng)纖維修復(fù)和再生周期較長。
[Abstract]:Purpose. To establish an animal model of trigeminal nerve injury in mice, to simulate the pathogenesis and outcome of neurotrophic keratitis; to observe the clinical changes and repair process of cornea in mice with trigeminal nerve removal; to explore the density of cornea nerve in mice with denervated innervation. The effect of corneal epithelial integrity and function. Materials and methods. Seventy adult male C57BL / 6 mice were randomly divided into 7 groups (10 mice in each group). The first branch of the left trigeminal nerve was removed by anhydrous ethanol method. Tear secretion tests, corneal fluorescein staining and confocal microscopy were performed on the normal control group and the experimental group at different time points, respectively. Then the mice were killed by intraperitoneal injection of excessive pentobarbital sodium, and the left cornea and intracranial trigeminal nerve branches were taken out respectively. The cornea nerves of mice were labeled with Tubulin 尾-鈪，

本文編號(hào)：1543505