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論文名稱(外文):Effects of Hyaluronan on the Gene Expression of Inflammatory Cells
指導教授(外文):Lynn L.H. Huang
外文關鍵詞:Wound HealingGrowth factorCytokineHyaluronan
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傷口癒合為人體受傷後皮膚組織再生的過程,由非常複雜的生理機制調控。許多研究指出,傷口初期的發炎反應是啟動傷口癒合的關鍵,發炎細胞在其中扮演相當重要的角色,除了進行病原菌的清除,也會分泌生長因子及細胞激素來加強發炎並活化組織細胞的增生、分化和細胞外基質的生合成。研究顯示,透明質酸在傷口癒合中能防止發炎反應的過度發生且有促進傷口癒合的功能。因透明質酸具有高度的生物相容性、可分解吸收性、低免疫性及保水性,所以常被使用於傷口敷材的研發上。至於透明質酸對於發炎細胞的影響,尤其在基因表現上的影響,目前為止仍沒有相關的研究報導。本研究目的在探討透明質酸對於發炎細胞在增生、趨化及相關基因表現的影響,藉以推論透明質酸促進傷口癒合的機制。結果發現,在不同分子量與不同濃度的透明質酸處理下,中、高分子量(780、1470、2000、2590 kDa)的透明質酸能促進發炎細胞的增生與趨化反應,且增加HB-EGF、FGF-7、PDGF、VEGF、TGF-β、MMP家族等生長因子及細胞激素基因的表現。因此從結果推論,在傷口癒合初期給予透明質酸,可驅使發炎細胞遷移至傷口處,且刺激其增生,並提升發炎細胞在生長因子及細胞激素基因的表現量,進而達到促進傷口癒合的效果。
Wound healing can be divided into the process of coagulation, inflammation, tissue proliferation, and wound remodeling, each stage of the process not only overlap each other, and by the very complex regulation of physiological mechanisms. Known in the early wound healing, the inflammatory cells such as neutrophil and macrophage are playing an important role to eliminate the invader but also trigger the healing process.
By secreting the specific growth factors and cytokines to activate the downstream cell (including fibroblasts, keratinocytes and epidermal cells) proliferation and differentiation, then to promote extracellular matrix synthesis to assist wound healing.
Extracellular matrix components - hyaluronan can prevent the excessively inflammatory response in the wound healing process. Because hyaluronan has a highly biocompatible, biodegradable absorbent, low immunogenicity, and the excellent water retentivity. Often used in the research and development of a wound dressing materials. There are supporting cells to migration and resist the mechanical stress. As regards the influence of hyaluronan for the inflammatory cells, especially in the influence on the gene expression, by far not related to the study reported. So the purpose of this study was to explore hyaluronan for cell proliferation, cell chemotaxis and gene expression and mechanisms in order to infer hyaluronan to promote wound healing.
The results showed that the treatment of the different molecular weights and different concentrations of hyaluronan, the high molecular weight of hyaluronan(780, 910, 1470, 2000, 2590 kDa) can promote the proliferation and chemotaxis of inflammatory cells, and increased HB-EGF, FGF-7, PDGF, VEGF, TGF-β, MMP family (Collagenase, MT-MMP) and other growth factor and cytokine gene expression. We conclude that the early stages of wound healing give extra hyaluronan, by trigger inflammatory cells migrate to the wound, and stimulate their proliferation and inflammatory cells in growth factor and cytokine gene expression, thus achieving promote wound healing. Since we understand the expression profile between extracellular matrix and inflammatory cell, it facilitating future development of related medical products industry to accelerate wound healing and can achieve the effect without scars.

中文摘要 1
Abstract 2
誌謝 3
目錄 4
表目錄 7
圖目錄 8
符號及名詞縮寫 10
一、研究背景與目的 13
1.1 傷口癒合 13
1.1.1 影響傷口癒合之因子 14
1.2 發炎細胞簡介 14
1.2.1 免疫細胞概述 14
1.2.2 發炎細胞與傷口癒合 15
1.3 細胞外基質簡介 15
1.3.1 細胞外基質在傷口癒合之研究 15
1.3.2 透明質酸(hyaluronan)概述 16
1.3.3 透明質酸與發炎反應之關係 17
1.4 生長因子、細胞激素以及基質金屬蛋白酶簡介 18
1.4.1 生長因子(growth factor) 18
1.4.2 細胞激素(cytokine) 19
1.4.3 基質金屬蛋白酶(matrix metalloproteinase,MMP) 20
1.5 研究目的 21
二、研究材料與方法 22
2.1 實驗藥品 22
2.2 實驗儀器與材料 24
2.3 實驗設計 27
2.3.1 發炎細胞之鑑定 27
2.3.2 發炎細胞分析技術之建立 27
2.4 實驗方法 29
2.4.1 發炎細胞之鑑定 29
2.4.2 發炎細胞分析技術建立 31
三、研究結果 41
3.1 實驗結果 41
3.1.1發炎細胞之鑑定 41
3.1.2 發炎細胞分析技術之建立 42
四、討論 50
4.1 實驗討論 50
4.1.1 發炎細胞之增生試驗 50
4.1.2 發炎細胞之基因表現量測定 51
五、參考文獻 78
六、附錄 82
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