【摘要】：研究目的視網膜色素變性(retinitis pigmentosa,RP)是視網膜變性中最常見的一組以進行性光感受器細胞及視網膜色素上皮(retinitis pigment epithelium,RPE)功能喪失為共同表現的遺傳性疾病。而原發(fā)性視網膜色素變性合并視網膜血管進行性閉塞是近年來眼科醫(yī)師逐步認識的一種疾病。國內曾以原發(fā)性血管閉塞性視神經視網膜病變做個案報道,強調不明原因,原發(fā)性血管閉塞為特點而國外未見報道。在進一步完善資料的補充搜集之后,本研究對此病的臨床特點進行總結,并對部分患者進行既往未曾做過的全基因組外顯子測序以試圖闡明其發(fā)病機制并印證既往我們對此病的認識。研究方法回顧性分析23例(46只眼)原發(fā)性RP合并視網膜血管閉塞患者的臨床資料。其中男性12例,女性11例;年齡從6～70歲平均49.96±15.80歲�；颊呔p眼受累。患者均進行了包括眼底檢查、熒光素眼底血管造影、視網膜電圖等檢查。采集其中6個患者的血樣,還采集了其中1個患者父母(無臨床表現)血樣,共8個樣本。進行全基因組外顯子測序。測序主要包括目標區(qū)域序列的富集、DNA測序、生物信息學統(tǒng)計三個主要步驟。測序結果中對出現在2個及以上case的位點進行了分析,重點關注沒有注釋到單核苷酸多態(tài)性數據庫的位點,可能是潛在的與RP相關的新的位點;進一步關注篩選突變類型為錯義突變等能直接引起氨基酸改變的位點,這樣功能變化會相對比較明顯。這樣大概只有不到300個位點,81個基因;跟進位點depth≥10且quality≥20進行過濾。然后對這些潛在位點應用京都基因和基因組百科全書(Kyoto Encyclopedia of Genesand Genomes,KEGG)和基因本體論數據庫(gene ontology,GO)進了功能富集分析。并結合臨床背景、經驗,進行綜合篩選過濾,尋找潛在的位點和致病基因。研究結果原發(fā)性RP合并視網膜血管閉塞患者的臨床表現為眼前節(jié)均無炎性反應,眼壓正常,晶狀體混濁20只眼,未見虹膜新生血管;患者眼底視乳頭色淡,其中蒼白如月者17只眼;視網膜血管變細,鼻側血管更明顯,血管旁可見白鞘。由于患者病程不一,其視網膜血管閉塞程度亦不同,嚴重者血管可閉塞成白線。視網膜色素紊亂和脫色素點,無典型骨細胞樣色素沉著。未見視網膜新生血管,無增生性視網膜病變及玻璃體積血。20只眼眼底熒光血管造影示視乳頭始終呈低熒光或晚期淡熒光充盈,局限于有光感眼或無光感眼。視乳頭部位有一小段血管有熒光素充盈或完全無血管充盈者17只眼。整個眼底可見椒鹽狀熒光素充盈。吲哚青綠造影檢查表現為脈絡膜血管較細,熒光素充盈較弱,退行較快,視網膜血管閉塞表現同熒光素眼底血管造影所見。視網膜電圖檢查顯示晚期明視、暗視ERG的a,b波幅為無波型或近無波型�；颊叨嘤幸姑な�。8個樣本全基因組外顯子測序結果出現了已報道的RP相關致病基因的單核苷酸多態(tài)性位點,又在一些基因的編碼區(qū)發(fā)現了新的突變位點,并導致了氨基酸改變。基因注釋發(fā)現與組織凋亡,神經褪變,抑制細胞增殖等相關。應用KEGG和GO數據庫尋找生物學相關通路,顯示存在有凋亡通路,神經軸突導向通路,血管內皮生長因子信號通路,視黃醇代謝通路等。結論本組研究患者眼底均表現為視乳頭色淡,視網膜血管變細,廣泛視網膜色素上皮受累,可見脫色素斑點及有或無細小色素點;ERG檢查a、b波均為無波型或近無波型,支持本組原發(fā)性RP合并視網膜血管閉塞患者屬于原發(fā)性RP(毯層RP)范疇。同時也有其自身臨床特點:1.晚期血管可完全閉塞成白線或近完全閉塞,視神經明顯萎縮可以蒼白如月。2.視網膜色素上皮以進行性萎縮為主、無典型骨細胞樣色素增殖沉著,未見視網膜新生血管及玻璃體出血。3.此病病程進展速度似較原發(fā)性RP為快。全基因組外顯子測序結果中出現了已報道的RP致病基因的單核苷酸多態(tài)性位點,是否這些位點在疾病中高發(fā)并與RP相關有待驗證。在一些基因編碼區(qū)發(fā)現了引起氨基酸改變的很多未見報道的新突變位點,基因注釋發(fā)現與組織凋亡,神經褪變,抑制細胞增殖等相關。并有基因參與凋亡、神經軸突導向、血管內皮生長因子信號等生物學通路。若從理論上完全解釋還有待進一步的工作來證實。
[Abstract]:Retinitis pigmentosa (RP) is one of the most common genetic diseases in the degeneration of retina, which is characterized by progressive photoreceptor cells and retinal pigment epithelium (RPE). Primary retinal pigment degeneration and progressive occlusion of retinal vessels are a disease which has been gradually recognized by the ophthalmologist in recent years. A case report of primary angiitis obliterans optic neuropathy was reported in China. It was emphasized that it was not reported in foreign countries due to unknown reasons and primary vascular occlusion. After further improvement of the data collection, this study summarized the clinical features of the disease and sequenced all genome exons that had not been previously done by some patients in an attempt to elucidate the pathogenesis of the disease and confirm our knowledge of the disease. Methods The clinical data of 23 cases (46 eyes) with primary RP combined with retinal vessel occlusion were analyzed retrospectively. Among them, 12 males and 11 females were females, and the average age ranged from 6 to 70 years, and the average age ranged from 49. 96 to 15. 80 years. Both eyes of the patient were involved. All patients underwent examinations including fundus examination, fluorescein angiography, electroretinogram, etc. Blood samples were collected from six of these patients, and a total of 8 samples were collected from one of the patients (no clinical presentation). All genome exons were sequenced. The sequencing mainly includes three main steps: enrichment of target region sequence, DNA sequencing and bioinformatics statistics. The results of sequencing showed that the sites of 2 and more cases were analyzed, and the sites that were not annotated to the single-point polymorphism database might be potential new sites related to RP. Further attention is paid to screening mutation types such as missense mutation and so on, which can directly cause amino acid changes, so that the functional changes can be relatively obvious. Thus, there are only less than 300 sites, 81 genes, a follow-up site depth column 10, and a quality table 20 for filtering. These potential sites were then subjected to functional enrichment analysis using Kyoto Encyclopedia of Genesand Genes (KEGG) and Gene Ontology Database (GO). Combined with the clinical background and experience, the comprehensive screening and filtration were carried out to find the potential sites and pathogenic genes. Results The clinical manifestations of primary RP-combined retinal vascular occlusion were no inflammatory reaction in anterior segment, normal intraocular pressure, 20 eyes with posterior lens opacity, and no iris neovascularization. The blood vessels at the nose side are more obvious, and white blood vessels can be seen near the blood vessels. Because the course of the patient is not one, the degree of retinal blood vessel occlusion is different, and the blood vessel of the serious person can be blocked into white line. Retinal pigment disorder and depigmentation point, no typical osteoclast-like pigmentation. No retinal neovascularization, no proliferative retinopathy and vitreous hemorrhage were seen. 20 eyes with fundus fluorescein angiography showed that the nipple was always low or late fluorescent filling, limited to light-sensitive eyes or no light-sensitive eyes. There is a small segment of blood vessel at the papillary site with fluorescein filling or no blood vessel filling 17 eyes. The whole fundus of the fundus can be filled with salt-like fluorescein. Angiographic examination showed that the choroid vessels were thinner, the fluorescein filling was weak, the regression line was faster, and the retinal vessel occlusion was seen with fluorescein angiography. The examination of electroretinogram showed that the amplitude of a and b of dark ERG was wave-free or nearly wave-free. The results showed that there was a single polymorphic site of the reported RP-related pathogenic gene, and a new mutation site was found in the coding region of some genes, which led to amino acid change. Gene annotation has been found to be related to tissue apoptosis, neurodegeneration, inhibition of cell proliferation, and the like. The KEGG and GO databases were used to find biological correlation pathways, and there were apoptotic pathways, neurite-directed pathways, vascular endothelial growth factor signaling pathways, retinal metabolism pathways, and the like. Conclusion The fundus of the patients in this group is characterized by pale nipple color, thinning retinal vein, extensive retinal pigment epithelium involvement, visible depigmentation spot and or without fine pigment spot, ERG examination a and b wave are wave-free or near-wave type, The patients with primary RP combined with retinal vascular occlusion belong to the category of primary RP (blanket layer RP). At the same time, it has its own clinical features: 1. Late blood vessels may be completely occluded into white lines or near complete occlusion, and the optic atrophy may pale as pale as month. Retinal pigment epithelium is mainly composed of progressive atrophy, no typical osteoclast-like pigment proliferation, no retinal neovascularization and vitreous hemorrhage. The progress of the disease progression seems to be faster than that of the primary RP. The results of all-genome exon sequencing showed the reported single-site polymorphic sites of RP-pathogenic genes, and whether these sites were high in the disease and associated with RP-related needs to be verified. In some gene coding regions, many novel mutation sites have been found to cause amino acid changes, and gene annotation has been found to be related to tissue apoptosis, neurodegeneration, inhibition of cell proliferation, and the like. and has genes involved in biological pathways such as apoptosis, neurite outgrowth, vascular endothelial growth factor signaling, and the like. If the theory is fully explained and the work to be further worked out, it is proved.
【學位授予單位】：天津醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R774.1

【相似文獻】

相關期刊論文 前10條

1 鄭軍,孫佰山;視網膜出血48例臨床分析[J];職業(yè)與健康;2003年10期

2 楊艷麗,周炳華;老年性視網膜出血145例病因分析[J];長春中醫(yī)學院學報;2004年04期

3 Goldman M.;Dagan Z.;Yair M. ;朱新菊;;嬰幼兒嚴重咳嗽與視網膜出血[J];世界核心醫(yī)學期刊文摘(兒科學分冊);2006年11期

4 倪德明;中醫(yī)治療常見視網膜出血四法[J];江西中醫(yī)藥;1994年S2期

5 魏桂英，，徐克;輸血后視網膜出血一例[J];中華眼底病雜志;1994年04期

6 本刊編輯部;第5例報告——自發(fā)性視網膜出血[J];中華眼底病雜志;2001年03期

7 王溪;王致和醫(yī)師對視網膜出血癥的治療體會[J];蘭州醫(yī)學院學報;2003年03期

8 王永宏;王春芳;閆建林;王芳;;山西太原機關工作人員視網膜出血的影響因素流行病學調查[J];中國當代醫(yī)藥;2012年08期

9 黃肖梅;;中西醫(yī)結合治療視網膜出血的療效觀察[J];海峽藥學;2012年03期

10 鄭毓琳;鄭魁山;李志明;;針刺治療41例視網膜出血的初步觀察[J];中醫(yī)雜志;1957年06期

相關會議論文 前10條

1 楊艷麗;王奇;沃娜;路欣宇;;老年性視網膜出血145例病因分析[A];中華醫(yī)學會第十二屆全國眼科學術大會論文匯編[C];2007年

2 王錫夫;李峰;張健;張傳記;孫洪然;;中藥黃斑復原湯對視網膜挫傷的療效觀察[A];第三次全國中醫(yī)、中西醫(yī)結合眼科學術交流會論文匯編[C];2003年

3 張京紅;;視網膜中央動靜脈阻塞臨床治療觀察[A];中國眼底病論壇·全國眼底病專題學術研討會論文匯編[C];2008年

4 包中余;王玲;;挫傷性視網膜出血中西醫(yī)結合治療38例臨床觀察[A];浙江省第十二屆農村醫(yī)學暨鄉(xiāng)鎮(zhèn)衛(wèi)生院管理學術會議論文匯編[C];2004年

5 陳芳;糜漫天;;牛磺酸對大鼠視網膜缺氧適應調節(jié)的影響[A];首屆中國西部營養(yǎng)與健康、亞健康學術會議論文集[C];2005年

6 姜健麗;羅丹;;中醫(yī)藥治療視網膜靜阻塞38例臨床小結[A];第三次全國中醫(yī)、中西醫(yī)結合眼科學術交流會論文匯編[C];2003年

7 姚紅艷;;放射性視神經病變[A];浙江省中西醫(yī)結合學會眼科專業(yè)委員會第十一次學術年會資料匯編[C];2008年

8 馬向明;陳麗欣;;視網膜挫傷32例眼底熒光血管造影分析[A];2006年浙江省眼科學術會議論文集[C];2006年

9 郟從賢;符忠;黃亮錫;;視網膜鈍挫傷的生物力學研究[A];第五次全國法醫(yī)學術交流會論文集[C];1996年

10 張志芳;李慶生;潘琳;;不同中醫(yī)內治法對實驗性家兔視網膜血管病變的干預研究[A];第六屆全國中醫(yī)中西醫(yī)結合眼科學術交流會論文匯編[C];2007年

相關重要報紙文章 前10條

1 李水銀;糖尿病視網膜出血的治療[N];中國中醫(yī)藥報;2002年

2 朱有章;全身用藥也可能傷視力[N];大眾衛(wèi)生報;2009年

3 婁底市中醫(yī)院眼科 羅建國 副主任醫(yī)師;胎頭受壓易致視網膜出血[N];大眾衛(wèi)生報;2001年

4 高彥彬;糖尿病視網膜病變[N];健康報;2003年

5 韓詠霞;云南白藥輔治視網膜出血[N];農村醫(yī)藥報（漢）;2004年

6 張明亮;哪些全身疾病要查眼底[N];家庭醫(yī)生報;2008年

7 張世節(jié);警惕年齡相關性黃斑變性[N];家庭醫(yī)生報;2006年

8 上海市第十人民醫(yī)院眼科 王方 博士;莫讓“三高”毀了你的心靈之窗[N];上海中醫(yī)藥報;2009年

9 尤志軍;小心藥物 害你眼[N];大眾衛(wèi)生報;2003年

10 李麗云;不要忽視藥物對眼睛的損害[N];中國中醫(yī)藥報;2003年

相關博士學位論文 前10條

1 虞竣;糖尿病引起的小鼠視網膜神經節(jié)細胞生理活動的變化[D];復旦大學;2013年

2 曹平;糖尿病微血管并發(fā)癥視網膜血管管徑的變化及芪明顆粒干預的影響[D];成都中醫(yī)藥大學;2016年

3 王明揚;原發(fā)性視網膜色素變性與視網膜血管閉塞的相關性研究[D];天津醫(yī)科大學;2014年

4 張杰;骨髓間充質干細胞分化為視網膜結構及其在重度視網膜損傷中的治療作用[D];中國人民解放軍軍事醫(yī)學科學院;2003年

5 余玲玲;“光復湯”對治療性視網膜激光光損傷修護作用的觀察[D];成都中醫(yī)藥大學;2008年

6 趙亮亮;視網膜鐵代謝與年齡相關性黃斑變性實驗研究[D];吉林大學;2015年

7 劉勇;人胚胎視網膜感光前體細胞移植治療視網膜色素變性的臨床前研究[D];第三軍醫(yī)大學;2009年

8 于磊;視網膜自適應光學成像質量的改進研究[D];中國科學院研究生院(長春光學精密機械與物理研究所);2015年

9 張世杰;Genistein影響視網膜趨化因子表達及血—視網膜屏障損害的實驗研究[D];復旦大學;2004年

10 李世迎;人胚胎視網膜移植到光誘導視網膜變性豬的視網膜下腔的實驗研究[D];第三軍醫(yī)大學;2006年

相關碩士學位論文 前10條

1 王永宏;常規(guī)體檢中年齡≥45歲人員視網膜出血的影響因素的研究[D];山西醫(yī)科大學;2012年

2 任韜冉;超廣角眼底熒光血管造影對視網膜中央靜脈阻塞的重新認知[D];河北醫(yī)科大學;2015年

3 顧倬;高海拔缺氧條件下HIF-1α在大鼠視網膜的表達與視網膜厚度變化的相關性研究[D];蘭州大學;2015年

4 徐軍;SD-OCT視網膜圖像神經纖維層的檢測與研究[D];南京理工大學;2017年

5 沈煒;轉化生長因子-β在正常和糖尿病大鼠視網膜中的基因表達研究[D];第二軍醫(yī)大學;2004年

6 曹丹;轉化生長因子-βⅠ型受體和Ⅱ型受體在正常及糖尿病大鼠視網膜中的基因表達研究[D];第二軍醫(yī)大學;2006年

7 劉攀;高危新生兒視網膜出血相關因素分析[D];中南大學;2013年

8 師燕蕓;兔眼局限性視網膜轉位術的實驗研究[D];山西醫(yī)科大學;2003年

9 鄧君;不同方法將骨髓間充質干細胞移植到大鼠視網膜下的比較[D];南方醫(yī)科大學;2014年

10 李倩;醋酸曲安奈德及其溶媒對視網膜形態(tài)學影響的實驗研究[D];首都醫(yī)科大學;2007年

本文編號：2283852