多功能蛋白聚糖在瘢痕组织及成纤维细胞中的表达

发布时间：2018-02-28 21:19

本文关键词： 瘢痕成纤维细胞细胞外基质多功能蛋白聚糖转化生长因子β1　出处：《昆明医学院》2010年硕士论文　论文类型：学位论文

【摘要】： [目的]通过测定多功能蛋白聚糖(versican)在瘢痕组织及瘢痕成纤维细胞中的表达情况,探讨其在增生性瘢痕和瘢痕疙瘩形成中的作用,以及versican和TGF-β1的表达关系,为临床预防和治疗瘢痕提供病理学及基因水平的理论依据。 [方法](1)对正常皮肤及病理性瘢痕成纤维细胞进行体外培养,采用光镜观察、电镜检查对成纤维细胞形态、活性、凋亡,RNA提取分析,用实时荧光定量PCR检测veriscan以及其四种亚型和TGF-β1的mRNA的表达情况；(2)采用组织块石蜡切片对病理性瘢痕组织中的versican和TGF-β1进行免疫组化检测,并运用SPSS 17.0软件对使用数据进行t检验,通过以上实验观察分析,研究versican对增生性瘢痕、瘢痕疙瘩以及成纤维细胞生物学行为的影响及病理改变；(3)对照实时荧光定量PCR和免疫组化结果,分析versican和TGF-β1的表达关系。 [结果](1)光镜下与正常皮肤成纤维细胞相比,病理性瘢痕成纤维细胞排列极性消失,细胞体积较大,形态不规则；(2)扫描电镜下：病理性瘢痕成纤维细胞粗面内质网大量增多,并扩展成囊,胞质内微丝、微管增多,表明其合成蛋白及胶原纤维的功能活跃;(3)免疫组化提示病理性瘢痕组织切片中versican主要表达于胶原纤维及成纤维细胞中,而真皮组织中未见明显表达,病理性瘢痕组织中versican和TGF-β1表达量较正常皮肤明显升高,具有统计学意义(P0.05)；(4)实时荧光定量PCR检测结果示瘢痕疙瘩以及正常瘢痕成纤维细胞中versican及其四种亚型均有明显表达,瘢痕疙瘩versican和TGF-β1的mRNA表达均较正常瘢痕及正常皮肤成纤维细胞升高。 [结论]Versican及其四种亚型在瘢痕疙瘩及正常瘢痕成纤维细胞中有明显表达,可能对病理性瘢痕发生发展及其成纤维细胞的生长、增殖及胶原合成起促进作用；versican的表达与TGF-β1相关,共同促进病理性瘢痕的发生与发展。
[Abstract]:[objective] to investigate the expression of versican and TGF- 尾 1 in hypertrophic scar and keloid by detecting the expression of multifunctional proteoglycan in scar tissue and scar fibroblasts. To provide the theoretical basis of pathology and gene level for clinical prevention and treatment of scar. [methods] normal skin and pathological scar fibroblasts were cultured in vitro. The morphology, activity and apoptosis of fibroblasts were detected by light microscope and electron microscope. The expression of veriscan, its four subtypes and the mRNA expression of TGF- 尾 1 were detected by real-time fluorescence quantitative PCR. The tissue paraffin sections were used to detect versican and TGF- 尾 1 in pathological scar tissue by immunohistochemistry. T test was performed with SPSS 17.0 software. The effect of versican on the biological behavior of hypertrophic scar, keloid and fibroblasts and its pathological changes were studied. The expression of versican and TGF- 尾 1 were analyzed by real-time quantitative PCR and immunohistochemistry. [results] compared with normal skin fibroblasts under light microscope, pathological scar fibroblasts disappeared in polarity, larger in size and irregular in shape. Under scanning electron microscope, the rough endoplasmic reticulum of pathological scar fibroblasts increased significantly. The expression of versican in the sections of pathological scar tissues was mainly expressed in collagen fibers and fibroblasts, which indicated that the synthesis protein and the function of collagen fibers were active. However, the expression of versican and TGF- 尾 1 in pathological scar tissue was significantly higher than that in normal skin. The results of real-time fluorescence quantitative PCR analysis showed that versican and its four subtypes were significantly expressed in keloid and normal scar fibroblasts. The mRNA expression of versican and TGF- 尾 1 in keloid was higher than that in normal scar and normal skin fibroblasts. [conclusion] the expression of Versican and its four subtypes in keloid and normal scar fibroblasts may play an important role in the development of pathological scar and the growth of fibroblasts. The expression of TGF- 尾 1 may play a role in promoting proliferation and collagen synthesis in keloid and normal scar fibroblasts. To promote the occurrence and development 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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