Immunohistochemical studies on the role of glycosaminoglycans in thepathogenesis of corneal disorders

糖胺聚糖在角膜疾病发病机制中作用的免疫组织化学研究

• 批准号: 11671732
• 负责人: HIRANO Koji
• 金额: $ 2.11万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671732/
• 关键词: proteoglycans; glycosaminoglycans; collagen fibrils; keratan sulphate; condroitin sulphate; corneal transporency; グリコサミノブリカン; ケラタン硫酸; コンドロイチシ硫酸; コラーゲン細線維; 角膜の透明性

To clarify the surface topography of D-periodic collagen fibrils, we applied atomic force microscopy (AFM) to the collagen fibrils of mouse corneal stroma and sclera. AFM showed that the depth of the groove in the D-periodicity of corneal fibrils was shallow compared with that of the sclera. Ruthenium red (RR) method showed that proteoglycans (PGs) or glycosaminoglycans (GAGs) distributed on the surface of collagen fibrils in the cornea and sclera. The difference in ultrastructure between cornea and sclera could explain the differences in the number or the distribution of molecule in both tissues.In the other experiment, we compared the surface structure of human corneal collagen before and after the digestion with chondroitinase ABC or kertanase by using corneal samples obtained from autopsy. The surface structure did not changed after the chondroitinase ABC digestion where as the groove of the D-periodicity changed to be much deeper after keratanase.This shows that keratan sulphate associate PGs should depress the groove of the surface of human corneal collagen fibrils, and these PGs may act for the regular distribution of collagen fibrils, and thus, may contribute for maintaining the transparency of the cornea. We further investigated the distribution of PGs or GAGs during corneal wound healing by the immunohistochemical study by using the human corneal graft obtained during regrafting, and showed that chondroitin sulphate A had an important role during wound healing.

为了阐明D-周期性胶原纤维的表面形貌，我们应用原子力显微镜（AFM）的小鼠角膜基质和巩膜的胶原纤维。AFM显示角膜原纤维D周期沟的深度较巩膜浅。钌红（RR）法显示，角膜和巩膜中胶原纤维表面分布有蛋白多糖（PGs）或糖胺多糖（GAGs）。角膜和巩膜超微结构的差异可以解释两者在分子数量和分布上的差异。在另一个实验中，我们比较了软骨素酶ABC和角蛋白酶消化前后的人角膜胶原的表面结构。软骨素酶ABC消化后，表面结构没有改变，而角蛋白酶消化后，D周期的凹槽变深，说明硫酸角蛋白结合的PG抑制了人角膜胶原原纤维表面的凹槽，使胶原原纤维规则分布，从而有助于保持角膜的透明性。我们进一步研究了PG或GAG的分布在角膜伤口愈合过程中的免疫组织化学研究，使用在再移植过程中获得的人角膜移植，并显示硫酸软骨素A在伤口愈合过程中有重要作用。

项目成果

Miyagawa A, Kobayashi M, Fujita Y, Nakamura M, Hirano K et al: "Surface topology of collagen fibrils associated with proteoglycans in mouse cornea and sclera."Jpn J Ophthalmol. 44. 591-595 (2000)

Miyakawa A、Kobayashi M、Fujita Y、Nakamura M、Hirano K 等人：“与小鼠角膜和巩膜中的蛋白聚糖相关的胶原原纤维的表面拓扑。”Jpn J Ophamol。

Hirano K, Kojima T, Nakamura M and Hotta Y: "Triple Anterior Chamber after Full-Thickness Lamellar Keratoplasty for Lattice Corneal Dystrophy."Cornea. (In press).

Hirano K、Kojima T、Nakamura M 和 Hotta Y：“全层板层角膜移植术后三前房治疗格状角膜营养不良。”角膜。

溝口英里,平野耕治: "水疱性角膜症の経過中に細菌感染性角膜潰瘍を起こした2例"日本眼科紀要. 51. 837-840 (2000)

Eri Mizoguchi、Koji Hirano：“大疱性角膜病过程中细菌感染的角膜溃疡的两例”日本眼科通报 51. 837-840 (2000)。

Mizoguchi E, Hirano K: "Two cases of infections corneal ulcer in the course of bullous keratopathy"Ophthalmology (Jpn). 43. 183-186 (2001)

Mizoguchi E、Hirano K：“大疱性角膜病过程中感染性角膜溃疡的两例”眼科（日本）。

Miyagawa A,Kobayashi M,Fujita Y,Nakamura M,Hirano K et al: "Surface topology of collagen fibrils associated with proteoglycans in mouse cornea an sclera."Jpn J Ophthalmol. 44. 591-595 (2000)

Miyakawa A、Kobayashi M、Fujita Y、Nakamura M、Hirano K 等人：“与小鼠角膜和巩膜中的蛋白聚糖相关的胶原原纤维的表面拓扑。”Jpn J Ophamol。