臺灣博碩士論文加值系統

近年來對於抗生素的不當使用導致細菌具有抗藥性的機率逐年上升，然而抗生素開發的速度卻比不上細菌演化的速度，使得可以使用的抗生素正在減少中，但在第一個抗生素上市以前，就存在著使用光治療細菌感染的方法－光動力療法，光動力療法透過可見光與光敏劑反應後與將能量傳遞至周圍的氧分子，產生出具有破壞性的自由基，攻擊菌體細胞膜使其變形或分解，再者造成遺傳物質變異而達到殺菌的效果。
本研究使用波長為450nm至460nm之藍光進行光動力療法，搭配黃素單核苷酸、黃素腺嘌呤二核苷酸以及薑黃素三種具有吸收可見藍光波長能量的物質作為光敏劑，以上三種光敏劑分別在不同藍光能量的曝照下可對臨床上常見的抗藥性菌種革蘭氏陰性之綠膿桿菌以及革蘭氏陽性之金黃色葡萄球菌產生完全失活的效果。而光動力療法目前可行之治療對象為人體之上皮層部分，目前最常見的上皮感染則為青春痘之病原菌痤瘡丙酸桿菌，而在藍光光動力療法搭配上薑黃素，在藍光能量144焦耳時就可將痤瘡丙酸桿菌完全失活。
透過液相層析質譜儀的分析，可以看到光敏劑在光照之下產生了化學反應，同時在掃描式電子顯微鏡下也可觀察到菌體隨著光照時間的增加出現更嚴重的表面損傷，由此可以證明光敏劑確實可以增進光動力療法之療效，且進一步的釐清了光動能療法的抑菌機制。

The excessive and improper using of antibiotics makes more and more bacteria have carried the resistance of antibiotics which makes less available antibiotics. Researchers are investigating alternative anti-infective strategies to which it is supposed microorganisms will unable to develop resistance. Prominent among these strategies is a group of approaches which rely on light. One alternative way to use light as a therapy is photodynamic antimicrobial therapy (PDT). Photodynamic therapy relies on the interaction between harmless visible light, oxygen and photosensitizers to produce abundant destructive reactive oxygen species (ROS).
We choose the visible blue light (450nm-460nm), and select three photosensitizers Curcumin、FAD (Flavin adenine dinucleotide) and FMN (Flavin mononucleotide). After irradiate with different blue light energy, these photosensitizers can inactivation of G (+) bacteria S. aureus and G (-) bacteria P. aeruginosa. And inactivate the most common skin infection bacteria P. acnes by PDT combine with Curcumin. We found the most effective irradiation energy was 144 J
with P. acnes treated by PDT combine with curcumin.
In the end, we verified our result by Liquid Chromatography-Mass Spectrometry and Scanning Electron Microscope. In the total ion chromatogram, the intensity of product which comes from curcumin after blue light irradiation was increase with the time blue light irradiated. Under Scanning Electron Microscope, the bacteria cell wall appeared more damage when more blue light was irradiated.

目錄
致謝 I
中文摘要 II
Abstract III
目錄 IV

第一章 緒論 1
一、 光動力療法抑制致病菌之優勢 1
1.1、 抗生素療法之現況 1
1.2、 光動力療法 2
1.3、 光敏劑的選擇 3
1.4、 光敏劑之光化學反應 4
二、 光動力療法於抗菌之應用 5
2.1、 光動力療法之優勢 5
2.2、 光動力療法於微生物治療之應用 5
2.3、 痤瘡丙酸桿菌簡介 6
2.4、 皮癬菌病簡介 6
三、 液相層析質譜鑑定 6
3.1、 液相層析 6
3.2、 電噴灑游離法 7
第二章 研究方法及步驟 9
一、 實驗藥品 9
二、 儀器裝置 9
三、 菌株培養與收集 10
四、 菌量測定 10
五、 藍光光動力療法抑制菌體活性實驗 10
六、 液相層析質譜儀分析 11
第三章 實驗結果與討論 12
一、 藍光光動力療法對病原菌之殺菌效果評估 12
二、 藍光光動力療法搭配FMN對皮膚病原菌之殺菌效果評估 12
三、 藍光光動力療法搭配薑黃素對皮膚病原菌之殺菌效果評估 13
四、 藍光光動力療法搭配FAD對皮膚病原菌之殺菌效果評估 14
五、 藍光光動力療法與綠光光動力療法之療效比較 14
六、 Curcumin 之藍光化學反應 15
七、 電子顯微鏡觀察藍光光動力療法對於痤瘡桿菌之菌體影響 15
第四章 結論 17
第五章 圖表附錄 19
第六章 參考文獻 28

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