光化學動力療法對創傷弧菌在體內及體外之抗菌效果

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Title:	光化學動力療法對創傷弧菌在體內及體外之抗菌效果 In vitro and in vivo antimicrobial effect of photodynamic therapy on Vibrio vulnificus
Authors:	王尹怡 Yin-Yi Wang
Contributors:	莊銀清楊竹茂嘉南藥理科技大學：生物科技研究所
Keywords:	創傷弧菌革蘭氏陰性菌光化學動力療法甲苯胺藍 V. vulnificus Gram-negative in vivo Photodynamic therapy Toluidine blue O(TBO)
Date:	2005
Issue Date:	2008-10-31 16:14:40 (UTC+8)
Abstract:	創傷弧菌是一種具有獨特性致毒因子的，革蘭氏陰性菌、呈現桿狀且嗜鹽性的海洋弧菌。此細菌可使人類產生嚴重的皮膚軟組織感染及嚴重的菌血症。在體外試驗中，當細菌給予較低光能的照射(100 J/cm2、80 mW/cm2)及較低劑量的TBO (0.1 µg/ml)濃度時即能被完全殺死。在本研究中證實光化學動力療法能夠破壞細菌的細胞壁、降低細菌的活動力並降低其蛋白酶活性，而且光化學動力療法在毒殺細菌能力上與TBO的濃度及照射光的能量都具有線性關係，這些作用機制，部分是經由基態氧的作用而產生毒殺效果。另外，在體內試驗中，利用創傷弧菌以傷口感染的方式去感染老鼠，並以光化學動力療法和TBO治療並觀察其抗菌的效果。結果有53% (10/19)的老鼠存活下來，這些存活下來的老鼠是經過100 µg/ml的TBO及曝照在紅色光源中(150 J/cm2、80 mW/cm2)並曾經受過全身性感染而且感染的菌落數是一半致死劑量(LD50)的100倍。此試驗首次證實光化學動力療法能治療受創傷弧菌感染傷口的老鼠。經由這些結果，期望未來在臨床治療細菌感染上除了使用一般的化學治療外也能將此種治療方式作為另一個輔助療法，但此治療方式進入臨床前應再進一步的進行臨床試驗。 Vibrio vulnificus, a particularly virulence species of vibrio, is a gram-negative, rod-shaped, halophilic marine bacterium. In this thesis, though which the soft tissues of human skin could be severely infected and often cause fetal sepsis we have studied the antibacterial effects of toluidine blue O (TBO)-mediated photodynamic therapy (PDT) of on the wound with V. vulnificus infections in mice. In vitro, the bacteria were killed after exposure to a lower light intensity (100 J/cm2 at 80 mW/cm2) in the presence of low dose of TBO (0.1 µg/ml). PDT was found to severely damage the cell wall, markedly reduced cell motility and virulence. The cell-killing effects were dose-dependent on TBO concentration and light intensity, which were mediated partly through reactive oxygen species (ROS) and were generated during photodynamic reaction. In vivo, the mice were infected with a bacterial inoculation count even more 100 times LD50 and septicemia had been established. We found that 53% of the infected mice were survived after treatment with 100 µg/ml of TBO and irradiated with a broad spectrum red light (150 J/cm2, 80 mW/cm2). In conclusion, our study has demonstrated that PDT accompanying with TBO treatment can cure mice with otherwise fatal V. vulnificus wound infections. These promising results suggest the potential of this regimen as a possible alternative to antibiotics in future clinical applications.
Relation:	校內外完全公開
Appears in Collections:	[Dept. of Biotechnology (including master's program)] Dissertations and Theses

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