半乳糖β-1,4-糖苷键在瘢痕疙瘩组织中的定位与表达

中华皮肤科杂志 ›› 2013, Vol. 46 ›› Issue (7): 475-479.

半乳糖β-1,4-糖苷键在瘢痕疙瘩组织中的定位与表达

杨圣菊¹,孟国梁²,陈晓栋³

1. 南通大学附属医院
2. 南通大学
3. 南通大学附属医院皮肤科

收稿日期:2012-09-21 修回日期:2012-11-07 出版日期:2013-07-15 发布日期:2013-07-01
通讯作者: 陈晓栋 E-mail:dermatochen@163.com
基金资助:
应用比较蛋白质组学技术探讨非家族性瘢痕疙瘩的发病机制;应用比较蛋白质组学技术探讨非家族性瘢痕疙瘩的发病机制;糖基化终末产物在糖尿病大鼠皮肤纤维化中的作用

Location and expression of galactose β-1,4-glycosidic bonds in keloid tissue

Received:2012-09-21 Revised:2012-11-07 Online:2013-07-15 Published:2013-07-01
Contact: Chen Xiao-Dong E-mail:dermatochen@163.com

摘要/Abstract

摘要： 目的探讨半乳糖β-1,4-糖苷键在瘢痕疙瘩组织中的合成、定位及糖蛋白半乳糖基化在瘢痕疙瘩形成机制中的作用。方法 Lectin 印迹法观察瘢痕疙瘩、增生性瘢痕和正常皮肤组织中糖蛋白的糖基化水平；饱和苦味酸-天狼猩红偏振光法观察组织学结构及胶原的形态与分布。分析组织中Ⅰ、Ⅲ型胶原的比例；蓖麻凝集素-Ⅰ免疫荧光组化法分析组织中半乳糖β-1,4-糖苷键的表达与定位，并与Ⅰ型前胶原α1共定位。结果经糖链染色，与正常皮肤组织相比，瘢痕疙瘩组织在约30 000和40 000 处糖蛋白的半乳糖β-1,4-糖苷键表达增高。偏振光显微镜显示，瘢痕疙瘩组织中含有大量Ⅰ型胶原纤维，约占（71.53 ± 4.03）%，Ⅰ、Ⅲ型胶原比例为2.56 ± 0.53，高于正常皮肤组织（P < 0.05）。免疫荧光发现，半乳糖β-1,4-糖苷键均匀分布在瘢痕疙瘩组织成纤维细胞胞膜和胞质内，与正常皮肤组织相比其表达量明显增加，且与Ⅰ型前胶原α1存在共定位。结论瘢痕疙瘩组织中存在着半乳糖β-1,4-糖苷键的表达变化，且主要定位于成纤维细胞，提示半乳糖β-1,4-糖苷键可能参与瘢痕疙瘩修复过程中纤维过度增生相关因子的修饰调控。【关键词】瘢痕疙瘩；半乳糖β-1,4-糖苷键；胶原Ⅰ型；蓖麻蛋白

关键词: 瘢痕疙瘩, 半乳糖β-1,4-糖苷键, 蓖麻蛋白, 胶原Ⅰ型

Abstract: YANG Sheng-ju*, MENG Guo-liang, CHEN Xiao-dong. *Department of Dermatology and Venereology, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu, China Corresponding author: CHEN Xiao-dong, Email: dermatochen@163.com 【Abstract】 Objective To observe the expression and location of galactose β-1,4-glycosidic bonds （Gal β-1,4-GlcNAc） in keloid tissue, and to investigate the role of glycoprotein galactosylation in the formation of keloid. Methods This study included 10 keloid tissue specimens, 7 hyperplastic scar tissue specimens, and 6 normal skin specimens. Lectin blot analysis was performed to measure the glycosylation level of glycoproteins, saturated picric acid - Sirius red staining followed by polarization microscopy to observe the type, expression and distribution of collagens in these specimens. The type I/type III collagen ratio was calculated. Immunofluorescence-based histochemistry was carried out by using Ricinus communis agglutinin I to analyze the expression and location of Gal β-1,4-GlcNAc in these skin samples, and double immunofluorescent staining to observe the colocalization of Gal β-1,4-GlcNAc and type Ⅰprocollagen α1. Results Compared with the normal skin tissue, the keloid tissue showed a significant increase in the expression of Gal β-1,4-GlcNAc in glycoproteins with a relative molecular mass of 30 000 to 40 000. Polarization microscopy revealed that there was a considerable expression of typeⅠcollagen fibers, which amounted to （71.53 ± 4.03）% in all the collagen fibers. The type I/type III collagen ratio was significantly higher in keloid tissue specimens than in normal tissue specimens （2.56 ± 0.53 vs. 0.91 ± 0.11, P < 0.05）. Fluorescence microscopy showed that Gal β-1,4-GlcNAc was uniformly distributed in the membrane and cytoplasm of fibroblasts in keloid tissue, and the expression intensity of Gal β-1,4-GlcNAc in keloid tissue was notably stronger than that in normal skin tissue. There was a colocalization between Gal β-1,4-GlcNAc and typeⅠprocollagen α1 in keloid tissue. Conclusions The expression of Gal β-1,4-GlcNAc, which is mainly observed in fibroblasts, is upregulated in keloid tissue, suggesting that Gal β-1,4-GlcNAc may be involved in the modulation of factors responsible for excessive fibre formation during the repair process of keloid. 【Key words】 Keloid; Galβ-1,4GlcNAc; Collagen type Ⅰ; Ricin

Key words: Galβ-1,4GlcNAc

中图分类号:

R751

杨圣菊孟国梁陈晓栋. 半乳糖β-1,4-糖苷键在瘢痕疙瘩组织中的定位与表达[J]. 中华皮肤科杂志, 2013, 46(7): 475-479.

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