臺灣博碩士論文加值系統

本研究主要目的為，探討發光二極體(Light Emitting Diode，LED)光照刺激對於高血糖鼠燙傷傷口中轉化生長因子β1(transforming growth factor - β1，TGF-β1)分泌量的影響。使用Wistar鼠做為實驗動物，隨機分成4個組別。高血糖鼠的誘導會先將老鼠禁食18小時後，採腹腔注射方式(intraperitoneal，i.p.)注射Alloxan monohydrate (80~ 100 mg/kg)，於第4、8天測量血糖，兩次血糖值都大於300 mg/dL後才開始進行後續實驗。燙傷傷口製造會使用加熱至100~110℃的黃銅塊接觸背部皮膚20秒，面積大小為1×1cm2。實驗所使用的LED波長為630 nm，功率密度14 mW/cm2，照射時間280秒，總能量為4 J/cm2，兩天照光一次。評估方式採用組織學分析，以及使用ELISA量測TGF-β1的含量。根據記數的的結果發現，正常鼠經過LED光照刺激後，其巨噬細胞和纖維母細胞最大值分別在第8天和第14天出現，比未照光的正常鼠提前兩天出現。而高血糖鼠經過LED光照刺激後，其巨噬細胞最大值出現在第8天，同樣也比未照光的高血糖鼠提早2天出現。而高血糖鼠照光組的纖維母細胞最大值出現在第14天，比未照光組的第18天提前了4天。在血管生成的部分，正常鼠和高血糖鼠經過LED光照刺激後，血管的最大值都同樣提前到第14天出現，比未照光組的第16天提早了2天。而在TGF-β1檢測的結果發現，正常鼠經過LED光照刺激後對於TGF-β1的分泌沒有明顯的效果，兩組之間幾乎沒有任何的差異。而在高血糖鼠的部份，高血糖鼠傷口的TGF-β1始終維持在很高的濃度，但是經過LED光照刺激後TGF-β1濃度會在第14天以後開始減少。由以上的結果顯示，LED光照刺激可使正常鼠和高血糖鼠傷口巨噬細胞、纖維母細胞和血管生成的最大值提前出現，並且可以讓高血糖鼠傷口TGF-β1濃度降低。 因此，我們得出以下結論，LED可使正常鼠傷口癒合時間縮短至少兩天，並且可以使高血糖鼠傷口癒合末期的TGF-β1濃度降低，進而促進傷口癒合。

The aim of this study was to determine effects of LED light irradiation on the secretion of TGF-β1 from the burns of hyperglycemic rats. We used Wistar rats for our experimental animals. All rats were devided into six groups. Hypergiycemic rats were induced with a single intraperitoneal dose of alloxan monohydrate (80~100 mg/kg) after an 18 hour overnight fast.The blood glucose was tested on 4 and 8 days after induced and blood glucose level more than 300 mg/dL were selected for the further study. The burned wound was created on the back of rats using a metal rod preheated to 100~110℃. Each wound size was 1×1cm2. The LED was given with these parameters: 630 nm of wavelength, 14 mW/cm2 of power density, 4 J/cm2 of energy density for 280 seconds, and once for two days.The wound healing were evaluated by histological analysis and the secretion of TGF-β1. According to the results of cell count, in the normal rats, LED could make the maxima of macrophage and fibroblast appear early two days. In the hypergiycemic rats, LED made the maxima of macrophage appear early two days, and the maxima of fibroblast appear early four days. In the part of angiogenesis, our result showed that LED could accelerate the rate of angiogenesis and made the maxima of angiogenesis appear early two days. The result of TGF-β1assay showed that the LED seem to have no significant influence on normal rats. In hyperglycemic rats, LED could reduce the concentration of TGF-β1 in the last phase of wound healing. Our result showed that LED could make the maxima of macrophage, fibroblast and vessel early appeared, and reduce the concentration of TGF-β1in hyperglycemic rats. In conclusion, LED could make the healing time of normal rats early two days, and reduce the concentration of TGF-β1 on the last phase of wound healing in hyperglycemic rats to promote wound healing.

目錄
中文摘要…………………………………………………………………………I
Abstract…………………………………………………………………………III
誌謝…………………………………………………………………………………V
目錄………………………………………………………………………………VI
圖目錄……………………………………………………………………………VIII
第一章 緒論………………………………………………………………………1
1.1 前言……………………………………………………………………………1
1.2 研究動機和目的………………………………………………………3
第二章 文獻回顧…………………………………………………………………4
2.1 傷口癒合過程…………………………………………………………4
2.2 糖尿病對傷口癒合的影響……………………………………………………6
2.3 藥物誘發高血糖動物模式……………………………………………8
2.4 TGF-β1對傷口癒合的影響………………………………………………11
2.5 低能量雷射於傷口癒合的應用…………………………………………15
第三章 材料與方法…………………………………………………………………20
3.1 實驗儀設備………………………………………………………………20
3.2 實驗藥品…………………………………………………………………21
3.3 高血糖鼠的誘導…………………………………………………………22
3.4 燙傷傷口的製造…………………………………………………………23
3.5 實驗分組…………………………………………………………………24
3.6 LED光源設備與光刺激參數………………………………………………25
3.7 採樣與組織切片製作……………………………………………………26
3.8 TGF-β1含量檢測…………………………………………………………27
3.9 評估方式…………………………………………………………………28
3.10 統計方法……………………………………………………………29
第四章 實驗結果……………………………………………………………………30
4.1傷口外觀觀察…………………………………………………………30
4.2組織學分析……………………………………………………………33
4.3 TGF-β1檢測………………………………………………………………58
4.4 細胞與生長因子的關係…………………………………………………60
4.5 LED光照刺激對傷口癒合的影響…………………………………………63
4.6 高血糖對傷口癒合的影響………………………………………………64
4.7 實驗結果回顧……………………………………………………………68
第五章 結果討論…………………………………………………………………69
第六章 結論…………………………………………………………………………72
參考文獻…………………………………………………………………………73

圖目錄
圖2.1傷口癒合過程示意圖………………………………………………………5
圖2.2 Alloxan誘導糖尿病的流程………………………………………………10
圖2.3 TGF-β1對傷口癒合的影響………………………………………………14
圖3.1燙傷傷口圖示………………………………………………………………23
圖3.2 LED光源設備………………………………………………………………25
圖4.1 燙傷第0、2、4、6、8天傷口外觀……………………………………………31
圖4.2 燙傷第10、12、14、16、18、20天傷口外觀……………………32
圖4.3 正常鼠未燙傷皮膚組織……………………………………………………36
圖4.4 正常鼠未燙傷與燙傷後組織狀況…………………………………………37
圖4.5 正常鼠燙傷第8天組織狀況……………………………………………………38
圖4.6 正常鼠燙傷第10天組織狀況…………………………………………39
圖4.7 正常鼠燙傷第12天組織狀況…………………………………………40
圖4.8 正常鼠燙傷第14天組織狀況………………………………………………41
圖4.9 正常鼠燙傷第16天組織狀況………………………………………………42
圖4.10 正常鼠燙傷第18天組織狀況……………………………………………43
圖4.11 正常鼠燙傷第20天組織狀況……………………………………………44
圖4.12 正常鼠組織學分析結果………………………………………………………45
圖4.13 高血糖鼠未燙傷皮膚組織狀況………………………………………………48
圖4.14 高血糖鼠未燙傷與燙傷後組織狀況…………………………………………49
圖4.15 高血糖鼠燙傷第8天組織狀況…………………………………………50
圖4.16 高血糖鼠燙傷第10天組織狀況……………………………………………51
圖4.17 高血糖鼠燙傷第12天組織狀況……………………………………………52
圖4.18 高血糖鼠燙傷第14天組織狀況……………………………………………53
圖4.19 高血糖鼠燙傷第16天組織狀況……………………………………………54
圖4.20 高血糖鼠燙傷第18天組織狀況……………………………………………55
圖4.21 高血糖鼠燙傷第20天組織狀況……………………………………………56
圖4.22 高血糖鼠組織學分析結果…………………………………………………57
圖4.23 TGF-β1檢測結果……………………………………………………………59
圖4.24 正常鼠TGF-β1與其分泌細胞的關係……………………………………61
圖4.25 高血糖鼠TGF-β1與其分泌細胞的關係…………………………………62
圖4.26 燙傷組與高血糖燙傷組結果比較……………………………………………66
圖4.27 燙傷照光組與高血糖燙傷照光組結果比較…………………………67

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