半乳糖β-1，4-糖苷键在瘢痕疙瘩组织中的定位与表达  

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

作　　者：杨圣菊[1] 孟国梁[2] 陈晓栋[1] 

机构地区：[1]南通大学附属医院皮肤性病科,226001 [2]南通大学医学院药理学系

出　　处：《中华皮肤科杂志》2013年第7期475-479,共5页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81041076）;江苏省“六大人才高峰”第七批项目（2010-WS039）;南通科技项目（BK2011044）;南通大学自然科学基金（112015）

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Leetin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；饱和苦味酸一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d1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30000和40000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-1，4一糖苷键的表达变化，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。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 Galβ-1,4-GlcNAc and type Ⅰ procollagen 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 type Ⅰ collagen 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-GIcNAc in keloid tissue was notably stronger than that in normal skin tissue. There was a colocalization between Gal β-1,4-GlcNAc and type Ⅰ procollagen 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 re

关 键 词：瘢痕疙瘩 半乳糖β--1 4一糖苷键 胶原I型 蓖麻蛋白 

分 类 号：R622[医药卫生—整形外科]