陳秀男臺灣大學：漁業科學研究所詹于萱Chan, Yu-HsuanYu-HsuanChan2007-11-282018-07-062007-11-282018-07-062005http://ntur.lib.ntu.edu.tw//handle/246246/59367多醣體 (β-Glucan ) 可以活化嗜中性白血球、巨噬細胞、自然殺手細胞等多種免疫細胞，刺激免疫反應的發生以及抵禦細菌的感染。最近的研究中指出多醣體能直接活化人類的纖維母細胞促進膠原蛋白的生成，進而幫助傷口的癒合。為了進一步證明多醣體的效能，本實驗利用菇菌多醣體溶液 ( Mushroonβ-Glucan solution ) 及羥乙基纖維素（Hydroxyethyl Cellulose；HEC）製成菇菌多醣體凝膠，作為傷口敷料使用，以進行大白鼠背部全深度傷口的治療評估。 實驗分成兩部分，第一部份實驗：定期觀察傷口外觀癒合的面積，結果顯示Control組、Glucan組、HEC組及HEC＋Glucan組的傷口分別於手術後第29天、第23天、第23天及第19天癒合，在四組處理組中以 HEC＋Glucan組的傷口癒合速度的最快速，相較於Control組明顯提早了10天。第二部分實驗：利用肉眼以及組織切片觀察傷口組織的生長情形，發現Glucan組和HEC＋Glucan組的傷口部位出現明顯的發炎反應及較多的纖維母細胞，產生較多的膠原蛋白。此外，四組處理組中以HEC＋Glucan組的傷口組織生長情況最佳，不僅在表皮細胞的增生和移行、纖維母細胞的聚集、肉芽組織的生長、血管新生等方面效果最為顯著，在傷口癒合後的膠原纖維的排列情況，也最接近正常皮膚組織。因此，本實驗推論菇蕈多醣體凝膠對於傷口癒合確實具有良好的促進效果。β-Glucan has been reported to be able to induce immune response and antibacterial activities through stimulating of neutrophils, macrophages, nature killer cells, and various immunocytes. Moreover, β-Glucan may improve wound healing of experimental animals through activating fibroblast and consequently enhance collagen biothynthses in the skin. In the present study, wound dressing containing mushroom （1→3）-β-D-glucan or（1→3）,（1→6） -β-D-glucan incorporated with hydroxyethyl cellulose (HEC ), were used for the investigation on the recovery process of skin following full-thickness dorsal skin defect of Wistar rat. The study was evaluated by measurment of recovery wound area after surgery and treatment. The average wound healing time for control, Glucan, HEC and HEC +Glucan were 29 days, 23 days, 23 days and 19 days respectively. It is suggested that Glucan, HEC and HEC +Glucan may significantly decrease the wound healing time in the experiment rats when compared with the control rats. The superior results on wound healing was obtained in the experiment using HEC +Glucan. The results on histopathological observasion showed that when Glucan and HEC +Glucan were used an obvious inflammation and significant fibroblast collagen biothynthesis were observed. Futhermore, the results revealed that HEC +Glucan exhibited the best efficiency on wound healing among the three treatments. Experiment on histolopathological observation comfirmed that HEC+Glucan may enhance to the fibroblast proliferation, keratinocyte proliferation and migration, granulation tissue formation, angiogenesis, and collagen arrangement.論文目次............................................... Ⅰ 圖目錄................................................. Ⅲ 表目錄................................................. Ⅴ 中文摘要............................................... 1 英文摘要............................................... 2 第一章 前言............................................ 4 第二章 文獻回顧........................................ 6 2-1 皮膚的功能及結構................................... 6 2-2 皮膚傷口的癒合過程................................. 10 2-3 製作傷口敷料的動機................................. 19 2-4 HEC 的特性介紹.................................... 23 2-5 多醣體 (β- Glucan ) .........,..................... 25 2-5-1 多醣體的生理活性................................. 25 2-5-2 多醣體與傷口復原................................. 27 2-5-3 多醣體的發展近況................................. 30 2-6 菇蕈多醣體（Mushroom β- Glucan）................... 31 第三章 材料與方法...................................... 34 3-1 實驗動物........................................... 34 3-2 實驗分組........................................... 34 3-3 多醣體的成分與含量測定............................. 35 3-3-1 多醣體的成分.................................. 35 3-3-2 多醣體的含量測定.............................. 36 3-4 實驗試劑的製備..................................... 38 3-5 製作大白鼠背部皮膚傷口模型......................... 40 3-6 評估傷口癒合過程的面積變化......................... 45 3-7 觀察傷口癒合過程的皮膚病理組織切片................. 46 3-7-1 製作新生皮膚組織切片.......................... 46 3-7-2 H&E 染色法.................................... 47 3-7-3 Masson 氏三色染色法........................... 49 3-7-4 觀察傷口癒合過程的組織生長變化................ 51 3-8 統計分析........................................... 51 第四章 實驗結果........................................ 52 4-1 多醣體含量的測定結果............................... 52 4-2 傷口癒合過程的外觀面積變化結果..................... 53 4-3 傷口癒合過程的外觀觀察及影像紀錄................... 57 4-4 傷口癒合過程的組織生長情形......................... 63 4-5 傷口癒合過程的膠原纖維生長及分布情................. 75 第五章 綜合討論........................................ 82 第六章 結論.............. .......................... 89 第七章 參考文獻........................................ 92 附錄一................................................ 1001646689 bytesapplication/pdfen-US大白鼠傷口癒合菇蕈多醣體羥乙基纖維素(HEC)RatWoud healingMushroom β- GlucanHydroxyethyl cellulose ( HEC )otherhttp://ntur.lib.ntu.edu.tw/bitstream/246246/59367/1/ntu-94-R92B45025-1.pdf