【摘要】：目的翼狀胬肉是我國常見眼表疾病之一，本病尚無有效的藥物治療，手術(shù)切除是目前治療翼狀胬肉的主要方法，但手術(shù)后并發(fā)癥多且手術(shù)后的復(fù)發(fā)率很高，所以治療相當棘手，因此進一步研究翼狀胬肉的發(fā)病機制十分必要。本實驗旨在研究翼狀胬肉中COX-2與新生血管的表達情況，探討COX-2在翼狀胬肉新生血管生成和發(fā)展中的作用，為探索翼狀胬肉的靶向治療提供新的理論依據(jù)。 方法使用蘇木素-伊紅染色（HE染色）法及免疫組化的方法對實驗標本進行研究。第一步：臨床收集組織標本用于切片制作。門診手術(shù)獲得正常球結(jié)膜組織23例，翼狀胬肉組織45例，術(shù)前裂隙燈檢查排除參與者其他角結(jié)膜性疾病。所有組織經(jīng)過固定劑固定，濃度依次遞升的乙醇脫水，使用石蠟包埋，組織切片，并做好標記并保存?zhèn)溆�。第二步：HE染色法觀察翼狀胬肉與正常球結(jié)膜組織的形態(tài)學(xué)差異。取切片，經(jīng)脫蠟后蘇木素浸染5min，將細胞核染成藍色，鹽酸酒精分色后伊紅染色10s，切片脫水、透明后中性樹膠封片，顯微鏡下觀察。第三步：免疫組織化學(xué)方法分別檢測翼狀胬肉和球結(jié)膜組織中COX-2、VEGF、PDGF與CD31分布及表達情況。取切片，經(jīng)脫蠟后置于PH6.0的抗原修復(fù)液中高熱修復(fù)6min，3％去離子水封閉內(nèi)源性抗原，在不同組織編號的玻片上分別加入一抗，4℃孵育過夜，沖洗后加入對應(yīng)二抗，DAB顯色，顯微鏡下觀察并計數(shù)。 結(jié)果（1）HE染色后正常球結(jié)膜組織與翼狀胬肉組織中形態(tài)學(xué)表現(xiàn)：正常球結(jié)膜組織表層上皮細胞排列整齊，基底上皮細胞邊界清晰，間質(zhì)血管少；翼狀胬肉翼狀胬肉組織表現(xiàn)為上皮層及基質(zhì)淺層成纖維細胞增殖，，表層上皮細胞排列紊亂，細胞體積增大，基底上皮邊界模糊，間質(zhì)中新生血管大量增生。（2）免疫組織化學(xué)方法分別檢測翼狀胬肉和球結(jié)膜組織中COX-2、VEGF、PDGF與CD31分布及表達情況：23例球結(jié)膜組織中均為呈陰性，翼狀胬肉組織樣本中有36例COX-2表達陽性(P 0.01)； CD31在兩種組織中均有表達，通過計數(shù)及統(tǒng)計，翼狀胬肉組織中MVD為20.3±4.4，球結(jié)膜組織中MVD為9.7±2.8(P0.01)； COX-2表達陽性的翼狀胬肉組織中MVD為19.06±1.84，表達陰性的翼狀胬肉組織中MVD為10.44±2.98(P0.01)。翼狀胬肉組織切片中34例PDGF表達陽性，36例COX-2表達陽性的胬肉組織中32例PDGF陽性；翼狀胬肉組織切片中39例VEGF表達陽性，36例COX-2陽性的胬肉組織中34例VEGF表達陽性；差異有統(tǒng)計學(xué)意義（P0.005）。在促進翼狀胬肉新生血管生成過程中COX-2的表達與血管相關(guān)因子VEGF和PDGF具有相關(guān)性(r＝0.458, r＝0.621)。 結(jié)論本實驗研究發(fā)現(xiàn)翼狀胬肉組織中MVD、VEGF、PDGF以及COX-2的表達明顯強于正常球結(jié)膜組織，另外，COX-2可能協(xié)同VEGF及PDGF共同刺激翼狀胬肉新生血管的發(fā)生。以上結(jié)果不僅證實COX-2在翼狀胬肉的發(fā)病中發(fā)揮重要的作用，同時進一步證實翼狀胬肉屬于紫外線誘導(dǎo)相關(guān)腫瘤性疾病。
[Abstract]:Objective pterygium is one of the common ocular surface diseases in China. There is no effective drug treatment for pterygium. Surgical resection is the main method to treat pterygium at present, but there are many complications after operation and the recurrence rate after operation is very high, so the treatment is quite difficult. Therefore, it is necessary to further study the pathogenesis of pterygium. The purpose of this study was to investigate the expression of COX-2 and neovascularization in pterygium and to explore the role of COX-2 in angiogenesis and development of pterygium, and to provide a new theoretical basis for the targeted treatment of pterygium. Methods hematoxylin-eosin staining (HE staining) and immunohistochemical method were used to study the experimental specimens. Step 1: clinical tissue specimens are collected for slice making. 23 cases of normal bulbar conjunctiva and 45 cases of pterygium were obtained by outpatient operation. All tissues were fixed with fixants, then dehydrated by ethanol with increasing concentration, paraffin embedded, tissue sections, labeled and preserved. Step 2: the morphological difference between pterygium and normal bulbar conjunctiva was observed by HE staining. The nuclei were stained blue after dewaxing and stained with hematoxylin for 5 min. The nuclei were stained with eosin for 10 s after separation of hydrochloric alcohol. The distribution and expression of COX-2,VEGF,PDGF and CD31 in pterygium and bulbar conjunctiva were detected by immunohistochemistry. After dewaxing, 3% deionized water was placed in the antigenic repair solution of PH6.0 for 6 min to seal the endogenous antigen. The first antibody was added to the different tissue numbered glass slides, incubated overnight at 4 鈩

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