实时荧光定量聚合酶链式反应检测瘢痕疙瘩中核心蛋白多糖的表达  

Detection of decorin expression in keloid with fluorescent quantitative-PCR

作　　者：刘流[1] 赵娴[1] 赫佳[1] 袁瑞红[1] 赵德萍[1] 韩雪松[1] 

机构地区：[1]昆明医科大学第一附属医院整形外科,650032

出　　处：《中华医学美学美容杂志》2013年第4期287-290,共4页Chinese Journal of Medical Aesthetics and Cosmetology

基　　金：云南省自然科学基金资助项目（编号：2005c0072M）;云南省社会发展攻关项目（编号：2007CA006）

摘　　要：目的探讨瘢痕疙瘩成纤维细胞中核心蛋白多糖的表达、含量，及其在瘢痕疙瘩形成中的作用和机制。方法对瘢痕疙瘩、正常瘢痕以及正常皮肤成纤维细胞进行体外培养，采用光镜、透射电镜观察成纤维细胞形态、活性及凋亡；应用实时荧光定量聚合酶链式反应（FQ-PCR）对核心蛋白多糖以及B1转化生长因子（TGF-β1）的mRNA表达进行检测、分析。结果瘢痕疙瘩成纤维细胞形态不规则、排列紊乱，线粒体增多，粗面内质网扩张呈囊，细胞核常染色质丰富，表明其合成蛋白的功能活跃；瘢痕疙瘩成纤维细胞中核心蛋白多糖mRNA含量较正常瘢痕或正常皮肤成纤维细胞降低，而TGF-β1 mRNA表达则较正常皮肤及瘢痕组织成纤维细胞升高。结论核心蛋白多糖在瘢痕疙瘩成纤维细胞内含量较正常皮肤明显减少，提示其对成纤维细胞增殖、合成的抑制作用随之减弱，同时使TGF-β1表达上调，导致成纤维细胞的大量增生、迁移，合成过量胶原。表明核心蛋白多糖是抑制瘢痕疙瘩形成的重要因子。Objective To detect the expression and content of decorin in fibroblasts of keloid to deeply reveal the mechenism and the role of decorin plays in scar formation. Methods Fibroblasts of keloid, normal scar and normal skin were cultured in vitro, and the morphology, activity, apoptosis of fibroblast were observed under light microscope and electron microscope; the mRNAs of decorin and TGF-β1 were detected and analyzed with real-time fluorescent quantitative-PCR （FQ-PCR）. Results Fibroblasts of keloid showed irregular morphology, larger size and disorder arrangement. There were a large number of mitochondria, swelling rough endoplasmic reticulum, and euchromatin-rich in nucle- us of fibroblasts, suggesting the protein synthesis of keloid fibroblast was very active. Compared with normal skin, the expression of decorin was significantly lower in keloid fibroblast; On the contrary, the expression of TGF-β1 was significantly higher in keloid fibroblast than in normal scar and normal skin. Conclusions Compared with normal skin, the expression of decorin in keloid fibroblast is signif- icantly lower. Lower content of decorin in early stage of wound healing may induce weakly suppression of proliferation and synthesis of fibroblast, and up-regulate the activity of TGF-β1, which promotes the proliferation, migration and excessive collagen synthesis of the fibroblast of keloid. Thus, decorin is an suppressor factor of keloid formation.

关 键 词：瘢痕疙瘩 核心蛋白多糖 81转化生长因子 成纤维细胞 实时荧光定量聚 合酶链式反应 

分 类 号：R622[医药卫生—整形外科]