多功能蛋白聚糖在瘢痕組織及成纖維細胞中的表達
本文關(guān)鍵詞: 瘢痕 成纖維細胞 細胞外基質(zhì) 多功能蛋白聚糖 轉(zhuǎn)化生長因子β1 出處:《昆明醫(yī)學(xué)院》2010年碩士論文 論文類型:學(xué)位論文
【摘要】: [目的]通過測定多功能蛋白聚糖(versican)在瘢痕組織及瘢痕成纖維細胞中的表達情況,探討其在增生性瘢痕和瘢痕疙瘩形成中的作用,以及versican和TGF-β1的表達關(guān)系,為臨床預(yù)防和治療瘢痕提供病理學(xué)及基因水平的理論依據(jù)。 [方法](1)對正常皮膚及病理性瘢痕成纖維細胞進行體外培養(yǎng),采用光鏡觀察、電鏡檢查對成纖維細胞形態(tài)、活性、凋亡,RNA提取分析,用實時熒光定量PCR檢測veriscan以及其四種亞型和TGF-β1的mRNA的表達情況;(2)采用組織塊石蠟切片對病理性瘢痕組織中的versican和TGF-β1進行免疫組化檢測,并運用SPSS 17.0軟件對使用數(shù)據(jù)進行t檢驗,通過以上實驗觀察分析,研究versican對增生性瘢痕、瘢痕疙瘩以及成纖維細胞生物學(xué)行為的影響及病理改變;(3)對照實時熒光定量PCR和免疫組化結(jié)果,分析versican和TGF-β1的表達關(guān)系。 [結(jié)果](1)光鏡下與正常皮膚成纖維細胞相比,病理性瘢痕成纖維細胞排列極性消失,細胞體積較大,形態(tài)不規(guī)則;(2)掃描電鏡下:病理性瘢痕成纖維細胞粗面內(nèi)質(zhì)網(wǎng)大量增多,并擴展成囊,胞質(zhì)內(nèi)微絲、微管增多,表明其合成蛋白及膠原纖維的功能活躍;(3)免疫組化提示病理性瘢痕組織切片中versican主要表達于膠原纖維及成纖維細胞中,而真皮組織中未見明顯表達,病理性瘢痕組織中versican和TGF-β1表達量較正常皮膚明顯升高,具有統(tǒng)計學(xué)意義(P0.05);(4)實時熒光定量PCR檢測結(jié)果示瘢痕疙瘩以及正常瘢痕成纖維細胞中versican及其四種亞型均有明顯表達,瘢痕疙瘩versican和TGF-β1的mRNA表達均較正常瘢痕及正常皮膚成纖維細胞升高。 [結(jié)論]Versican及其四種亞型在瘢痕疙瘩及正常瘢痕成纖維細胞中有明顯表達,可能對病理性瘢痕發(fā)生發(fā)展及其成纖維細胞的生長、增殖及膠原合成起促進作用;versican的表達與TGF-β1相關(guān),共同促進病理性瘢痕的發(fā)生與發(fā)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.
【學(xué)位授予單位】:昆明醫(yī)學(xué)院
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2010
【分類號】:R363
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