發(fā)布時間：2019-07-02 09:29

【摘要】：青光眼是世界第二大致盲性疾病,臨床表現(xiàn)為進行性視野丟失及視乳頭凹陷。高眼壓是青光眼視神經(jīng)病變最主要的病因,機械損傷可最終造成RGC凋亡、視神經(jīng)萎縮,即青光眼視神經(jīng)病變。視乳頭是眼球結(jié)構(gòu)的一個“薄弱點”,由眼內(nèi)壓產(chǎn)生的機械力作用于視乳頭周圍的眼球底部,并在致密的鞏膜和相對薄弱的視乳頭的移行處被放大,故視乳頭處的視神經(jīng)對眼內(nèi)壓的變化更為敏感,是青光眼視神經(jīng)病變首先累及的位點。視神經(jīng)篩板及球后環(huán)視乳頭組織內(nèi)的彈性纖維賦予了組織良好的順應(yīng)性和可塑性,可一定程度地緩沖眼內(nèi)壓波動對視神經(jīng)產(chǎn)生的影響。彈性蛋白是構(gòu)成彈性纖維的主要成分,LOXL1是催化彈性蛋白聚合再生的關(guān)鍵酶,對維持彈性纖維穩(wěn)態(tài)至關(guān)重要。假性色素剝脫青光眼(PEXG)患者的眼壓難控制并且對青光眼視神經(jīng)病變較敏感,在患者的篩板組織內(nèi)發(fā)現(xiàn)了廣泛的彈性纖維降解現(xiàn)象。近幾年的研究發(fā)現(xiàn)：LOXL1的基因表達缺陷與高眼壓同為PEXG的重要危險因素。那么,這兩個因素對青光眼視神經(jīng)病變的進展是否具有協(xié)同作用呢?彈性蛋白是否是這二者共同的作用點呢?作用的機制又是怎樣的呢?為了探討以上問題,我們采用了Loxl1基因敲除小鼠(簡稱Loxl1-/-)作為研究載體,WT小鼠作為對照,利用前房注射微粒珠技術(shù)給予慢性高眼壓損傷,比較小鼠對損傷的易感性,觀察彈性蛋白在病變中的表現(xiàn)。 本研究是中美聯(lián)合培養(yǎng)博士的合作項目,得到了國家留學基金委和美國國立衛(wèi)生院的大力支持。研究的貢獻是：鞏固了微粒珠手術(shù)建立小鼠慢性高眼壓這一新技術(shù)的應(yīng)用；豐富了對Loxl1-/-小鼠眼部表型的研究；為PEXG的病因機制提供了一個新的研究思路。 本論文由四部分組成。第一章前言首先提到彈性纖維對維系組織順應(yīng)性和可塑性的至關(guān)重要,概述了彈性纖維的合成降解過程以及相關(guān)的病理制,強調(diào)彈性纖維穩(wěn)態(tài)的維持需要LOXL1的參與。而后提出彈性纖維參與了青光眼視神經(jīng)病變的生物力學機制,LOXL1缺陷使彈性纖維從穩(wěn)態(tài)轉(zhuǎn)為紊態(tài),與高眼壓一同成為PEXG的主要病因。最后對Loxl1-/-小鼠以及小鼠高眼壓模型的發(fā)展概況進行了簡要介紹。而后,提出本文的研究方向和研究特色。 第二章所述工作為選擇前房注射微粒珠的手術(shù)技術(shù)建立小鼠慢性高眼壓模型。本實驗各選用25只C57/BL6(野生型和)Loxl1-/-小鼠,前房注射2μl微粒珠(6μm)+3μl粘彈劑,以tonolab小鼠專用眼壓測量機監(jiān)測小鼠術(shù)后眼壓,并且觀察眼部體征,觀察時間為術(shù)后56天。微粒珠手術(shù)成功建立了小鼠慢性高眼壓模型,兩組實驗動物術(shù)后眼壓變化的總體特征是相近的,WT眼壓的波動相對較大。 第三章中,我們在56天觀察期滿后取材,測量術(shù)眼眼軸變化,評估術(shù)眼視乳頭凹陷程度,并且對RGC軸突進行計數(shù),觀察了視神經(jīng)內(nèi)的病理形態(tài),明確微粒珠手術(shù)所造成的慢性高眼壓在小鼠眼內(nèi)導致了類青光眼視神經(jīng)病變。并且我們發(fā)現(xiàn)：Lox11-/-小鼠的眼球的擴張程度更明顯,眼軸的增長呈現(xiàn)高度眼壓依賴性,視乳頭凹陷、RGC軸突的病變程度都比WT嚴重,可見Lox11-/-小鼠對慢性高眼壓損傷更敏感。 第四章工作利用免疫熒光染色和蛋白印跡學技術(shù)對彈性蛋白在慢性高眼壓下的表現(xiàn)進行了研究。我們發(fā)現(xiàn)：Lox11-/-小鼠眼球后極部及球后環(huán)視乳頭組織內(nèi)的彈性纖維結(jié)構(gòu)排列疏松,在慢性高眼壓損傷下存在明顯的彈性蛋白再生障礙及廣泛降解現(xiàn)象,同時眼內(nèi)單體彈性蛋白及彈性蛋白降解產(chǎn)物堆積,印證了形態(tài)學的改變。 研究認為：Lox11-/-小鼠眼內(nèi)LOXL1表達缺陷使其在慢性高眼壓損傷下合并出現(xiàn)彈性蛋白的再生障礙以及異常降解,眼球壁以及環(huán)視乳頭組織的順應(yīng)性和可塑性下降,使得Lox11-/-小鼠的視神經(jīng)更易受到機械性損傷,彈性蛋白降解產(chǎn)物累積并發(fā)揮生物活性,可能進一步加劇視神經(jīng)病變程度。
[Abstract]:Glaucoma is the second-largest blindness disease in the world. The clinical manifestation is progressive visual field loss and papilla depression. High intraocular pressure (IOP) is the main cause of the optic neuropathy of the glaucoma, and the mechanical injury can ultimately cause the apoptosis of RGC and the atrophy of the optic nerve, that is, the optic neuropathy of the glaucoma. if the nipple is a "weak point" of the eye structure, the mechanical force generated by the internal pressure of the eye acts on the bottom of the eye around the nipple and is enlarged at the transitional portion of the dense sclera and the relatively weak visual nipple, so that the optic nerve at the nipple is more sensitive to the change in the internal pressure of the eye, It is the first involved site of the optic neuropathy of the glaucoma. The elastic fibers in the optic nerve screen plate and the post-spherical ring-optic papilla tissue give the tissue good compliance and plasticity, and the effect of the internal pressure fluctuation on the optic nerve can be buffered to a certain extent. The elastic protein is the main component of the elastic fiber, and the LOXL1 is the key enzyme for catalyzing the polymerization and regeneration of the elastic protein, and is essential for maintaining the steady state of the elastic fiber. It is difficult to control the intraocular pressure in the patients with pseudopigmentation glaucoma (PEXG) and is more sensitive to the optic neuropathy of the glaucoma, and a wide range of elastic fiber degradation is found in the patient's sieve plate tissue. In recent years, it has been found that LOXL1 gene expression defect and high intraocular pressure are an important risk factor for PEXG. So, does the two factors have a synergistic effect on the progression of the glaucoma optic neuropathy? Is the elastin a common point of action for both? What is the mechanism of action? In order to study the above problems, we used the Loxl1 knockout mouse (Loxl1-/-) as the research carrier and WT mice as the control, and using the anterior chamber injection of the particle bead technique to give the chronic high intraocular pressure injury, and compared the susceptibility of the mouse to the injury, The expression of elastin in the lesion was observed. The research is a cooperation project of the joint culture of China and the United States, and is supported by the NSFC and the National Health Center of the United States. The contribution of the study is to consolidate the application of the new technique for establishing the chronic high intraocular pressure in mice by the microbead operation, and to enrich the research on the eye phenotype of the Loxl1-/-mouse, and to provide a new research for the etiological mechanism of the PEXG. Road. This paper is composed of four parts The first part of the first chapter is to mention the importance of the elastic fiber in the maintenance of the compliance and plasticity of the tissue. The process of the synthesis and degradation of the elastic fiber and the related pathological system are introduced, and the maintenance of the steady state of the elastic fiber is emphasized. The LOXL1 is emphasized. The results show that the elastic fiber is involved in the biomechanical mechanism of the optic neuropathy of the glaucoma. The LOXL1 defect changes the elastic fiber from the steady state to the low-pressure state, and becomes the main body of the PEXG with the high intraocular pressure. The development of the high intraocular pressure model of Loxl1-/-mouse and mouse was also reviewed. In this paper, the research direction and the research of this paper are put forward. To study the features of the study. In the second chapter, the operative technique for the selection of the anterior chamber injection of the microbeads was used to establish the mice's chronic In this experiment,25 C57/ BL6 (wild type and) Loxl1-/-mice were selected, and the anterior chamber was injected with 2. m The model of chronic high intraocular pressure in mice was successfully established after operation for 56 days. The overall characteristics of the changes of intraocular pressure in two groups of experimental animals were similar, and the wave of intraocular pressure in the two groups was similar. In the third chapter, we observed the changes of the eye-eye axis after the expiration of the 56-day observation, and evaluated the degree of the eye-to-eye degree of the eye, and the RGC axons were counted, and the visual and visual effects were observed. The internal pathological form, the chronic high intraocular pressure caused by the fine particle-bead operation resulted in the blue-green in the mouse's eye. Optic neuropathy was found. And we found that the degree of dilation of the eye of the Lox11-/-mouse is more pronounced, the growth of the eye is highly dependent on the ocular pressure, the degree of lesion of the RGC axon is more severe than that of the WT, and the Lox11-/-mice are chronic high. Chapter 4: Immunofluorescence and Western Blot in the Treatment of Chronic High-intraocular Pressure The results of this study were studied. We found that the elastic fiber structure in the posterior polar part of the eye and the post-spherical ring of the eye of the mouse was loose, and there were obvious elastic protein regeneration and extensive degradation under the condition of chronic high intraocular pressure, and the elastic protein and the elastic protein of the intraocular monomer. the degradation products of the sex protein are stacked, The changes of the morphology were confirmed. The study found that the expression of LOXL1 in the Lox11-/-mice caused the combination of the regeneration barrier and the abnormal degradation of the elastin under the injury of chronic high intraocular pressure, the eye wall and the group of the ring-optic papilla. The compliance and plasticity of the woven fabric are reduced, so that the optic nerve of the Lox11-/-mouse is more susceptible to mechanical damage, the degradation products of the elastin are accumulated and the biological activity is exerted,
【學位授予單位】：復旦大學
【學位級別】：博士
【學位授予年份】：2011
【分類號】：R775

【相似文獻】

相關(guān)期刊論文 前10條

1 陳倩;孫興懷;俞道義;郭文毅;余曉波;戴毅;金曉紅;;腦源性神經(jīng)營養(yǎng)因子及其受體酪氨酸激酶B在慢性高眼壓獼猴初級視皮層的表達[J];中國眼耳鼻喉科雜志;2007年05期

2 李志勇,劉巖,張廣慶;針刺降低兔慢性高眼壓機理的實驗研究[J];中國中醫(yī)眼科雜志;2003年04期

3 彭清華;活血利水法治療慢性高眼壓臨床體會[J];實用中醫(yī)藥雜志;1994年06期

4 蔡豐英;慢性高眼壓對兔眼視神經(jīng)的影響[J];中國實用眼科雜志;1995年02期

5 朱海;龍心光;陳少基;葉靜;周蔚;李志勇;詹宇堅;;針刺對慢性高眼壓家兔眼壓及前房角組織病理變化的影響[J];中國中醫(yī)眼科雜志;2008年04期

6 彭清華;活血利水治療慢性高眼壓的臨床體會[J];遼寧中醫(yī)雜志;1995年02期

7 宋宗明,崔守信,張德秀;兔眼慢性高眼壓下內(nèi)核層細胞改變[J];眼科研究;2000年02期

8 杜兆江,劉杰,楊新光,王百忍,鄺芳,段小莉;白細胞介素-1β對慢性高眼壓狀態(tài)下大鼠視網(wǎng)膜的損傷作用[J];第四軍醫(yī)大學學報;2005年02期

9 朱海;龍心光;吳海科;周蔚;楊雪艷;陳少基;葉靜;李志勇;詹宇堅;;針刺對慢性高眼壓家兔前房角組織超微結(jié)構(gòu)的影響[J];新中醫(yī);2009年11期

10 張兆康;倘孟瑩;滕月;張麗霞;;益精杞菊地黃顆粒對慢性高眼壓大鼠視網(wǎng)膜神經(jīng)節(jié)細胞凋亡的影響[J];中國中醫(yī)眼科雜志;2018年06期

相關(guān)會議論文 前4條

1 王影;唐由之;馮俊;巢國俊;;慢性高眼壓兔模型的實驗研究[A];中華中醫(yī)藥學會第五次眼科學術(shù)交流會論文匯編[C];2006年

2 張琪;李平華;;核轉(zhuǎn)錄因子NF—κB在兔慢性高眼壓視網(wǎng)膜上的表達及其與細胞凋亡的關(guān)系[A];中華醫(yī)學會第十二屆全國眼科學術(shù)大會論文匯編[C];2007年

3 楊新光;于敬妮;王百忍;;兔慢性高眼壓球后視神經(jīng)損害的超微結(jié)構(gòu)改變[A];中華醫(yī)學會第十二屆全國眼科學術(shù)大會論文匯編[C];2007年

4 段宣初;;中南大學湘雅二院青光眼基礎(chǔ)研究進展[A];中華醫(yī)學會第十二屆全國眼科學術(shù)大會論文匯編[C];2007年

相關(guān)博士學位論文 前6條

1 陳涵;Lox11基因敲除小鼠對慢性高眼壓損傷的易感性研究[D];復旦大學;2011年

2 崔馨;SD大鼠視網(wǎng)膜慢性高眼壓損傷差異表達基因的研究[D];第三軍醫(yī)大學;2006年

3 歸東梅;氧適應(yīng)基因的調(diào)控與大鼠慢性高眼壓視網(wǎng)膜損傷關(guān)系初步研究[D];中國醫(yī)科大學;2007年

4 聶慶珠;AG調(diào)控的信號轉(zhuǎn)導途徑在大鼠慢性高眼壓視網(wǎng)膜損傷中的保護作用[D];中國醫(yī)科大學;2007年

5 吳繼紅;慢性高眼壓視網(wǎng)膜神經(jīng)節(jié)細胞進行性死亡的線粒體機制[D];復旦大學;2013年

6 王楷迪;Sonic hedgehog信號通路在大鼠慢性高眼壓視網(wǎng)膜神經(jīng)節(jié)細胞損傷中的保護作用及機制研究[D];復旦大學;2010年

相關(guān)碩士學位論文 前10條

1 朱海;針刺對慢性高眼壓家兔前房角組織超微結(jié)構(gòu)的影響[D];廣州中醫(yī)藥大學;2008年

2 王秀青;兔實驗性慢性高眼壓視網(wǎng)膜損害相關(guān)基因的克隆及篩選[D];第三軍醫(yī)大學;2003年

3 王廣博;尿激酶對慢性高眼壓大鼠視網(wǎng)膜神經(jīng)節(jié)細胞凋亡的影響[D];新鄉(xiāng)醫(yī)學院;2013年

4 呂炳健;血管內(nèi)皮生長因子對大鼠慢性高眼壓狀態(tài)下視網(wǎng)膜神經(jīng)節(jié)細胞的保護作用[D];第四軍醫(yī)大學;2005年

5 梁靜;川芎嗪對慢性高眼壓大鼠視網(wǎng)膜神經(jīng)節(jié)細胞凋亡的影響[D];新鄉(xiāng)醫(yī)學院;2012年

6 劉玨;慢性高眼壓下兔視網(wǎng)膜組織蛋白質(zhì)組變化的初步研究[D];重慶醫(yī)科大學;2010年

7 李諾;白蒺藜皂苷對慢性高眼壓兔視網(wǎng)膜神經(jīng)節(jié)細胞及視神經(jīng)的保護作用[D];湖北中醫(yī)藥大學;2010年

8 王英爽;AQP4在慢性高眼壓大鼠視網(wǎng)膜中的表達[D];中國醫(yī)科大學;2007年

9 朱璇;腫瘤壞死因子受體在慢性高眼壓大鼠視網(wǎng)膜中的表達[D];復旦大學;2009年

10 陳嵐;糖原合酶激酶3β對青光眼視網(wǎng)膜神經(jīng)節(jié)細胞作用的研究[D];華中科技大學;2011年

，

本文編號：2508807