核心蛋白多糖在瘢痕组织及成纤维细胞中的表达及意义

发布时间：2018-02-22 11:30

本文关键词： 瘢痕核心蛋白多糖成纤维细胞创伤愈合　出处：《昆明医学院》2010年硕士论文　论文类型：学位论文

【摘要】： 一、论文核心蛋白多糖在瘢痕组织及成纤维细胞中的表达及意义【目的】对病理性瘢痕组织及成纤维细胞中Decorin的含量及表达进行检测,深入探讨并揭示Decorin对瘢痕的作用及机制,为临床预防和治疗瘢痕提供分子及病理学水平的理论依据。【方法】(1)对病理性瘢痕成纤维细胞进行体外培养,采用光镜、扫描电镜观察成纤维细胞形态、活性及凋亡等;提取细胞RNA,应用RT-PCR对Decorin、TGF-β1的mRNA表达进行检测分析。(2)将病理性瘢痕组织进行固定、包埋、切片,采用免疫组织化学染色对Decorin、TGF-β1进行检测。通过以上实验观察及统计学分析,以研究Decorin对病理性瘢痕成纤维细胞生物学行为的影响及病理改变,Decorin在病理性瘢痕发生发展中的作用。【结果】(1)光镜下:成纤维细胞为贴壁生长,长梭形或多边形,呈栅栏状排列;与正常皮肤成纤维细胞相比,病理性瘢痕成纤维细胞形态不规则、体积较大,细胞排列较乱;(2)电镜下:与正常皮肤对照组相比,瘢痕成纤维细胞线粒体增多,粗面内质网增多并肿胀、扩张呈囊状,胞质内微丝微管增多,细胞核内常染色质丰富,表明其合成蛋白的功能活跃;(3)RT-PCR结果显示:瘢痕疙瘩成纤维细胞中Decorin mRNA含量较正常瘢痕或正常皮肤成纤维细胞降低;瘢痕疙瘩成纤维细胞中TGF-β1 mRNA表达较正常皮肤及瘢痕组织成纤维细胞升高;(4)免疫组化结果显示:Decorin主要表达于正常皮肤真皮层细胞外基质中及胶原纤维表面,与正常皮肤相比,Decorin在瘢痕组织中表达量明显降低;而TGF-β1在瘢痕组织中表达量明显高于正常皮肤中,具有统计学意义(P＜0.05)。【结论】Decorin在病理性瘢痕组织及成纤维细胞内含量较正常皮肤明显减少,提示在创面愈合早期由于Decorin含量降低,使TGF-β1活性上调,其对成纤维细胞的刺激作用也随之增加,引起成纤维细胞的大量增生、迁移,并合成过量胶原,这可能是病理性瘢痕形成的一个重要因素。
[Abstract]:1. Expression and significance of core proteoglycan in scar tissue and fibroblasts. [objective] to detect the content and expression of Decorin in pathological scar tissue and fibroblasts, and to explore the effect and mechanism of Decorin on scar in order to provide theoretical basis for clinical prevention and treatment of scar. [methods] the fibroblasts of pathological scar were cultured in vitro. The morphology, activity and apoptosis of fibroblasts were observed by light microscope and scanning electron microscope. The mRNA expression of TGF- 尾 1 was detected by RT-PCR. The pathological scar tissue was immobilized, embedded, sectioned and detected by immunohistochemical staining, and the expression of TGF- 尾 1 was detected by immunohistochemical staining. To study the effect of Decorin on the biological behavior of fibroblasts in pathological scar and the role of Decorin in the pathogenesis and development of pathological scar. [results] under the light microscope, fibroblasts were adherent, fusiform or polygonal, arranged in palisade shape. Compared with normal skin fibroblasts, pathological scar fibroblasts were irregular in shape and larger in volume. Compared with the normal skin control group, the mitochondria of scar fibroblasts increased, the rough endoplasmic reticulum increased and swollen, dilated in cystic shape, the cytoplasmic microfilament increased, and the nucleus was rich in chromatin. The results of RT-PCR showed that the content of Decorin mRNA in keloid fibroblasts was lower than that in normal scar or normal skin fibroblasts. The expression of TGF- 尾 1 mRNA in keloid fibroblasts was higher than that in normal skin and scar tissue fibroblasts. Compared with normal skin, the expression of TGF- 尾 1 in scar tissue was significantly lower than that in normal skin, and the expression of TGF- 尾 1 in scar tissue was significantly higher than that in normal skin (P < 0.05). [conclusion] the content of Decorin in pathological scar tissue and fibroblast is significantly lower than that in normal skin. It is suggested that in the early stage of wound healing, TGF- 尾 1 activity is upregulated due to the decrease of Decorin content, and the stimulating effect of TGF- 尾 1 on fibroblast is also increased. The proliferation and migration of fibroblasts and the synthesis of excessive collagen may be an important factor in the formation of pathological scar.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363

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