發(fā)布時(shí)間：2018-03-15 00:08

  本文選題：角膜營(yíng)養(yǎng)不良　切入點(diǎn)：TACSTD2基因　出處：《鄭州大學(xué)》2010年博士論文　論文類型：學(xué)位論文

【摘要】： 目的：觀察膠樣滴狀角膜營(yíng)養(yǎng)不良和施奈德角膜營(yíng)養(yǎng)不良這兩種罕見的角膜基質(zhì)營(yíng)養(yǎng)不良的臨床表現(xiàn),研究患者角膜組織病理學(xué)及活體角膜共焦顯微鏡和眼前節(jié)光學(xué)斷層掃描的圖像特征,同時(shí)分析膠樣滴狀角膜營(yíng)養(yǎng)不良致病基因TACSTD2基因和施奈德角膜營(yíng)養(yǎng)不良致病基因UBIAD1基因的突變情況,研究這兩種角膜基質(zhì)營(yíng)養(yǎng)不良的分子遺傳學(xué)機(jī)制。 方法：收集獨(dú)立的膠樣滴狀角膜營(yíng)養(yǎng)不良家系3個(gè)和施奈德角膜營(yíng)養(yǎng)不良家系1個(gè),對(duì)4個(gè)家系中的4例角膜營(yíng)養(yǎng)不良患者行以新鮮角膜作為供體的深板層角膜移植術(shù),將患者病變角膜組織做病理組織學(xué)、透射及掃描電鏡檢查,并在術(shù)前對(duì)患者角膜分別進(jìn)行裂隙燈拍照、活體共焦顯微鏡和眼前節(jié)光學(xué)相干斷層掃描,同時(shí)從患病者及其家系成員的外周靜脈血中提取基因組DNA,用特異性引物分別將致病基因TACSTD2和UBIAD1基因行PCR擴(kuò)增,PCR產(chǎn)物純化后進(jìn)行雙向測(cè)序,設(shè)50例正常人作為對(duì)照,分析患者基因型的改變。 結(jié)果：膠樣滴狀角膜營(yíng)養(yǎng)不良患者角膜表面可以見到呈半球形隆起的灰白色膠滴狀結(jié)節(jié)；病變角膜片在剛果紅染色下顯示特征性的角膜上皮下紅染的淀粉樣物質(zhì)沉積,累及前部角膜基質(zhì),角膜上皮基底膜和前彈力層結(jié)構(gòu)消失；掃描電鏡下可見角膜表層上皮細(xì)胞之間間隙增大,透射電鏡下可見角膜上皮基底細(xì)胞的基底面出現(xiàn)大量釘突狀突起結(jié)構(gòu)并向下插入其下的淀粉樣沉積物中,沉積物的超微結(jié)構(gòu)呈典型的淀粉樣纖維外觀；活體共焦顯微鏡檢查見患者角膜表層上皮細(xì)胞間連接異常,細(xì)胞間有較大間隙,上皮基底細(xì)胞層次不清、胞體腫脹,上皮基底細(xì)胞層下可見無(wú)定形的高反光物質(zhì)沉積,累及前部基質(zhì)；在眼前節(jié)光學(xué)相干斷層掃描圖像上,患者的角膜病變主要位于上皮細(xì)胞層和前部基質(zhì),角膜表面凹凸不平,呈波浪樣,角膜基質(zhì)的膠原板層規(guī)則排列被破壞,基質(zhì)內(nèi)呈強(qiáng)弱不一致的回聲信號(hào)。施奈德角膜營(yíng)養(yǎng)不良結(jié)晶型患者右眼角膜中央?yún)^(qū)基質(zhì)層混濁,角膜上皮下有散在點(diǎn)狀結(jié)晶,左眼角膜中軸部的前基質(zhì)層內(nèi)有一環(huán)形或邊緣不規(guī)整的、略呈盤狀的黃白色混濁,混濁由無(wú)數(shù)細(xì)小、針狀結(jié)晶組成；施奈德角膜營(yíng)養(yǎng)不良無(wú)結(jié)晶型患者可見雙眼角膜中軸部可見一無(wú)結(jié)晶圓盤狀混濁；透射電鏡下可見角膜基質(zhì)淺層與Bowman層中有膽固醇結(jié)晶沉積；活體共焦顯微鏡檢查見結(jié)晶型患者角膜前彈力層及淺基質(zhì)層內(nèi)可見到大量點(diǎn)狀、散在分布的結(jié)晶物,無(wú)結(jié)晶型患者則未見到針狀或方形的膽固醇結(jié)晶物質(zhì)存在于角膜組織內(nèi)；在眼前節(jié)光學(xué)相干斷層掃描圖像上,施奈德角膜營(yíng)養(yǎng)不良患者的角膜病變主要位于前部基質(zhì),顯示為高信號(hào)強(qiáng)度的致密物質(zhì)。分子遺傳學(xué)的研究發(fā)現(xiàn)3個(gè)膠樣滴狀角膜營(yíng)養(yǎng)不良家系的患者TACSTD2基因分別出現(xiàn)c.526 576de151、C.509CA和c.356GA突變。在患者中突變以純合子存在,在患者父母和其他未患病的親屬中以雜合子存在,50例正常對(duì)照者中未發(fā)現(xiàn)TACSTD2基因突變；施奈德角膜營(yíng)養(yǎng)不良家系的患者UBIAD1基因出現(xiàn)c.292 GA突變,在患者中突變以雜合子存在,50例正常對(duì)照者中未發(fā)現(xiàn)UBIAD1基因突變。 結(jié)論：膠樣滴狀角膜營(yíng)養(yǎng)不良特征性的臨床表現(xiàn)是角膜表面顆粒狀隆起物伴基質(zhì)混濁,病理組織學(xué)特征是角膜上皮下的淀粉樣物質(zhì)沉積；施奈德角膜營(yíng)養(yǎng)不良具有結(jié)晶型和非結(jié)晶型兩種臨床表現(xiàn),病理組織學(xué)特征是角膜基質(zhì)內(nèi)的膽固醇結(jié)晶物沉積；我們首次報(bào)道了膠樣滴狀角膜營(yíng)養(yǎng)不良和施奈德角膜營(yíng)養(yǎng)不良的活體共焦顯微鏡及眼前節(jié)相干光學(xué)斷層掃描圖像特征,這兩種檢查可發(fā)現(xiàn)角膜營(yíng)養(yǎng)不良角膜內(nèi)異常物質(zhì)的沉積和細(xì)胞結(jié)構(gòu)的改變,在疾病的隨訪和手術(shù)療效評(píng)估中有重要意義；分子遺傳學(xué)的研究發(fā)現(xiàn)膠樣滴狀角膜營(yíng)養(yǎng)不良和施奈德角膜營(yíng)養(yǎng)家系的患者分別呈現(xiàn)TACSTD2和UBIAD1基因突變,基因突變分析為疾病的確診提供可靠的依據(jù)并有助于對(duì)這兩種罕見角膜基質(zhì)營(yíng)養(yǎng)不良發(fā)病機(jī)制的進(jìn)一步認(rèn)識(shí)。
[Abstract]:Objective: To observe the gelatinous drop like corneal dystrophy and Schneider corneal dystrophy of the two rare corneal stromal dystrophy clinical manifestation, image characteristics and in vivo confocal microscopy and anterior segment optical tomography corneal tissue of patients with pathological study, mutation analysis of UBIAD1 gene of colloid droplet corneal dystrophy gene TACSTD2 gene and Schneider corneal dystrophy genes at the same time, study the two kinds of corneal dystrophy molecular mechanism.
Methods: collect separate colloid droplets of corneal dystrophies and Schneider 3 corneal dystrophies 1, of 4 families in 4 cases of corneal dystrophy patients with fresh corneal as deep lamellar keratoplasty donor, the lesions in patients with corneal tissue pathological histology. Transmission and scanning electron microscopy, and in preoperative patients cornea were examined by slit lamp photograph, coherence tomography in vivo confocal microscopy and anterior segment optical, genomic DNA was extracted from peripheral venous blood from patients and their family members, using specific primers respectively will TACSTD2 gene and UBIAD1 gene amplified by PCR and the purified PCR products were sequenced, 50 cases of normal people as control, analysis of the patient's genotype.
Results: the colloid droplet corneal dystrophy corneal surface can see a white gelatinous drop like nodules hemispherical bulge; corneal lesions showed amyloid deposition characteristics of corneal subepithelial red stained with Congo red staining, involving the anterior corneal stroma, corneal epithelial basement membrane disappeared and Bowman structure; scanning electron microscope showed that the corneal surface between the epithelial cell gap increased, visible basal corneal epithelial basal cells surface under transmission electron microscopy, the emergence of a large number of spike like protrusions and down into amyloid deposits under it, the ultrastructure of sediments showed the typical appearance of amyloid fibrils; in vivo confocal microscopy in patients with corneal epithelial surface. The connection between abnormal cells, intercellular large gap, epithelial basal cells and poorly, cell body swelling, epithelial basal cell layer showed amorphous high reflective 鐗╄川娌夌Н,绱強(qiáng)鍓嶉儴鍩鴻川錛涘湪鐪煎墠鑺傚厜瀛︾浉騫叉柇灞傛壂鎻忓浘鍍忎笂,鎮(zhèn)ｈ，

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