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研究生:黃彥豪
研究生(外文):Yen-hao Huang
論文名稱:五-胺基酮戊酸誘發之光動力抗菌效益的探討
論文名稱(外文):5-Aminolevulinic acid induced photodynamic antimicrobial effect.
指導教授:蔡翠敏
學位類別:碩士
校院名稱:臺北醫學大學
系所名稱:生物醫學材料研究所
學門:工程學門
學類:生醫工程學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:68
中文關鍵詞:五-胺基酮戊酸喀紫質光動力治療動力抗菌效益
外文關鍵詞:δ-Aminolevulinic acidALAPorphyrinsPhotodynamic therapyPDTAntimicrobial effectCarbopol
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由於細菌對抗生素產生抗藥性,使醫學面臨著越來越沒有適當有效的抗生素來對抗治療微生物感染性疾病的窘困,找出新的抗微生物感染療法實為醫學界的當務之急。五-胺基酮戊酸(5-Aminolevulinic acid)為多數生物細胞可自行生成的物質,可在細胞內轉化成在適當波長的可見光照射下呈現光動力效應的吡喀紫質(Porphyrins),鑑於五-胺基酮戊酸在抗腫瘤上之光動力治療(Photodynamic therapy)的成效,本研究藉由科學文獻的分析及實際的實驗操作下,探討五-胺基酮戊酸所誘發的光動力抗菌效益。
本研究利用革蘭氏陽性菌之金黃色葡萄球菌(S. aureus)、與革蘭氏陰性菌之綠膿桿菌(P. aeruginosa)等菌,在添加五-胺基酮戊酸、賦形劑(Carbopol 971P;CP)後於370C培養1小時,再以波長630±5 nm的紅光照射,使用ELISA-reader及plate counting等實驗操作來觀察細菌生長情形,以驗證五-胺基酮戊酸之光動力抗菌效益。
實驗結果發現金黃色葡萄球菌及綠膿桿菌等菌在添加五-胺基酮戊酸在紅光照射下呈現生長抑制的情形,而且更在合併使用賦形劑(CP)時,細菌存活率呈現更明顯的下降情形。由此可證五-胺基酮戊酸所誘發的光動力效應確實具有抗菌效果,而賦形劑(CP)對五-胺基酮戊酸所誘發的光動力抗菌效應確實有所幫助。又對抗生素oxacillin具有半敏感性(intermediate)的金黃色葡萄球菌在添加五-胺基酮戊酸且在紅光照射下的抗生素感受性的試驗中(antibiotic susceptibility Testing)呈現敏感性(sensitive)的結果。
In the present study we examined the effects of the photodynamic antimicrobial action by various amounts of δ-aminolevulinic acid(ALA) on the microorganisms Staphylococcus aureus(S. aureus) and Pseudomonas aeruginosa(P. aeruginosa). This was performed by solely inducing the porphyrin biosynthesis pathway. All strains exponentially growing were incubated in nutrient broth with ALA at 370C in the dark for 1 hr and then irradiated with 630±5 nm red light. Incubation of ALA with these microorganisms did not slow down their growth. When ALA-induced photodynamic treatment(ALA-PDT) in light dose 162 J/cm2 was performed onto bacterial strains S. aureus and P. aeruginosa, decrease of bacterial viabilities in the orders of magnitudes 6.0 and 4.1, respectively was found with 1.0 mM ALA, and the total eradication could be achieved when ALA concentrations equal or higher than 2.5 mM was used. Furthermore, when ALA-PDT was combined with a model antibiotic and/or a macromolecular excipient (carbopol 971P;CP), the results of antimicrobes became even more promising. We observed that S. aureus been intermediately susceptible to oxacillin became sensitive to oxacillin when in combination with ALA-PDT, even became sensitive to CP only. Our results showed that the PDT effect was worth further investigation as a standard anti-infection treatment modality, and further combination of ALA-PDT with other antibiotics or excipients may result in even better anti-microbial effect.
英文摘要(i)
中文摘要(iii)
謝誌(v)
目錄(vii)
圖目錄(viii)
表目錄(x)
第一章:緒論(1)
第二章:文獻探討(4)
2.1 五-胺基酮戊酸光動力抗菌效益(4)
2.2 五-胺基酮戊酸(5)
2.3 微生物防治(7)
2.4 光動力效應(12)
第三章:材料與方法(22)
3.1 材料(22)
3.2 儀器(26)
3.3 方法(27)
3.4 實驗目的(29)
第四章: 實驗結果與討論(30)
4.1 實驗一:葡萄球菌濃度對應ELISA-reader的檢量線實驗(30)
4.2 實驗二:葡萄球菌的ALA誘發光動力抗菌效應(30)
4.3 實驗三:綠膿桿菌濃度對應ELISA-reader的檢量線實驗(33)
4.4 實驗四:綠膿桿菌的ALA誘發光動力抗菌效應(33)
4.5 實驗五:賦形劑CP對ALA誘發光動力抗菌效應的影響(36)
4.6 實驗六:抗生素oxacillin合併ALA誘發光動力作用的抗菌效果(39)
第四章:結論與建議(40)
第五章:參考文獻(43)
第六章:附錄(45)
圖附錄(45)
表附錄(66)
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2.Jori G, Fabris C, Soncin M, Ferro S, Coppellotti O, Dei D, Fantetti L, Chiti G, Roncucci G. Photodynamic therapy in the treatment of microbial infections: basic principles and perspective applications. Lasers Surg Med. 2006 Jun;38(5):468-81.
3.Raab O. Uber die wirkung fluoriziender stoffe auf infusorien. Zeit Biol 1900;39;524-546
4.von Tappeiner H. Zur kenntis der lichtwirkenden(fluoreszierender) stoffe. Dtsch Med Wochen 1904;1:579-580
5.Kennedy JC, Pottier RH, Pross DC. Photodynamic therapy with endogenous protoporphyrin IX: basic principles and present clinical experience. J Photochem Photobiol B 1990;6:143–8.
6.Haydée Fukuda, Adriana Casas, Alcira Batlle. Molecules in focus Aminolevulinic acid: from its unique biological function to its star role in photodynamic therapy. The International Journal of Biochemistry & Cell Biology 37 (2005) 272–276
7.LiWei Ma﹐Saulius Bagdonas﹐Johan Moan. The photosensitizing effect of the photoproduct of protoporphyrin Ⅸ. J. Photochem. Photobiol.﹐B﹐2001;60:108-113.
8.Yeshayahu Nitzan, Mali Salmon-Divon, Einav Shporen and Zvi Malik. ALA induced photodynamic effects on Gram positive and negative bacteria. Photochem. Photobiol. Sci.;2004:3, 430–435
9.Reinhard Sailer , Wolfgang S.L. Strauss, Karsten Kiinig, Angelika Riick, Rudolf Steiner. Correlation between porphyrin biosynthesis and photodynamic inactivation of Pseudomonas aeruginosa after incubation with 5-aminolaevulinic acid. J. Photochem. Photobiol.﹐B﹐1997;39:236-242.
10.Lee CF, Lee CJ, Chen CT, Huang CT.δ-Aminolaevulinic acid mediated photodynamic antimicrobial chemotherapy on Pseudomonas aeruginosa planktonic and biofilm cultures. J. Photochem. Photobiol. B. 2004 Jul 19;75(1-2):21-5.
11.Hui-Chun Chen, Tsuimin Tsai, Design of a 5-Aminolevulinic Acid Delivery System for Photodynamic Diagnosis of Oral Premalignant and Malignant Lesions. 2002. Thesis Master of Sceince Grsduate Institute of Biomedical Materials, Taipei Medical University.
12.Jian-ming Li, Ormond Brathwaite,1 Sharon D. Cosloy,2 and C. S. Russell3. 5-Aminolevulinic Acid Synthesis in Escherichia coli. Journal of bacteriology, May 1989, p. 2547-2552
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15.George P. Tegos and Michael R. Hamblin1, Phenothiazinium Antimicrobial Photosensitizers Are Substrates of Bacterial Multidrug Resistance Pumps. Antimicrobial agent and chemotherapy, Jan. 2006, p. 196–203
16.Michael R. Detty, Scott L. Gibson, and tephen J. Wagner, Current Clinical and Preclinical Photosensitizers for Use in Photodynamic Therapy. Journal of medicinal chemistry. 2004, Volume 47, Number 16.
17.Z. Malik, J. Hanania and Y. Nitzan, Bactericidal effects of photoactivated porphyrins–an alternative approach to antimicrobial drugs, J. Photochem. Photobiol., B, 1990, 5, 281–93.
18.Theodrore M. Brody, Joseph Larner, Kenneth P. Human pharmcology: molecular to clinic.Third edition, 1998
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22.Mrinalini Sharma. Harsha Bansal and Pradeep Kumar Gupta, Photodynamic inactivation of antibiotic resistant strain of Pseudomonas aeruginosa by porphyrins induced by δ-aminolaevulinic acid, Indian J Med Res 116, September 2002, pp 99-105
23.K. Szocs, F. Gabor, G.. Csik, J. Fidy, δ-Aminolaevulinic acid-induced porphyrin synthesis and photodynamic inactivation of Escherichia coli B. J. Photochem. Photobiol. B:Biol. 50(1999)8-17
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