The study of in vitro and in vivo fermentation of bacteria in the skin microbiome

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施芯蒂(Shinta Marito) 查詢紙本館藏

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生命科學系

論文名稱

(The study of in vitro and in vivo fermentation of bacteria in the skin microbiome)

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摘要(中)

中文摘要
在皮膚上有許多不同微生物群存在，包括了細菌與真菌。菌與真菌的相互作用在人體中許多部份包括口腔和腸胃道都能找到。這樣物種間不同的交互作用在常見的丙酸桿菌屬能夠被觀察到。在正常皮膚與青春痘影響的的皮膚上，不同品系之丙酸桿菌分布有顯著的不同，這也顯示出不同品系間交流的可能性。紅色毛癬菌 (108 CFU/ml)與皮膚共生表皮葡萄球菌 (108 CFU/ml)在含有甘油條件下共同培養發現，能夠藉由甘油發酵並且產生短鍊脂肪酸。我們發現表皮葡萄球菌能夠干擾紅色毛癬菌生長。在丙酸痤瘡桿菌與表皮葡萄球菌共同培養的情況下，丙酸痤瘡桿菌的生長會被表皮葡萄球菌藉由蔗糖發酵之短鏈脂肪酸給抑制。在體內的實驗中我們發現，在老鼠耳朵的動物模式中表皮葡萄球菌藉由蔗糖發酵能夠改善丙酸痤瘡桿菌誘導的巨噬細胞發炎蛋白-2 (MIP-2)產生。此外，利用丙酸痤瘡桿菌之單株抗體能顯著的…減少老鼠耳朵模式的發炎反應及丙酸痤瘡桿菌之菌落。

關鍵字: 紅色毛癬菌、表皮葡萄球菌、丙酸痤瘡桿菌、短鏈脂肪酸、單株抗體

摘要(英)

Abstract

The skin is colonized by a diverse array of microorganisms including bacteria and fungi. The bacterium-fungal interaction is found in many parts of the human body including the oral cavity and gastrointestinal tract. Interaction between species of one common genus is observed in propionibacteria. The distribution of different Propionibacterium strains is significantly different on the skin of healthy subjects in comparison to acne-affected skin indicating a possible communication between bacterial strains. Co-culturing the Trichophyton rubrum (T. rubrum) (108 CFU/ml) with commensal Staphylococcus epidermidis (S .epidermidis) (108 CFU/ml) in presence of glycerol showed the production of short chain fatty acids (SCFAs) by fermentation of glycerol. We found that S. epidermidis interfered with the growth of T. rubrum. In co-culture of S. epidermidis and Propionibacterium acne (P. acnes), P. acnes growth was inhibited by production of SCFAs via the sucrose fermentation of S. epidermidis. In vivo studies showed that S. epidermidis sucrose fermentation can ameliorate the P. acnes-induced macrophage inflammatory protein-2 (MIP-2) production in the mouse ear. Additionally, P. acnes colonization and inflammation of mouse ear were significantly reduced by using monoclonal antibodies to P. acnes.

Keywords: T. rubrum, S. epidermidis, P. acnes, SCFA, monoclonal antibody.

關鍵字(中)

★ 紅色毛癬菌
★ 表皮葡萄球菌
★ 丙酸痤瘡桿菌
★ 短鏈脂肪酸
★ 單株抗體

關鍵字(英)

★ T. rubrum
★ S. epidermidis
★ P. acnes
★ SCFA
★ monoclonal antibody

論文目次

Table of Contents
中文摘要………………………………………………………………………… i
Abstract……………………………………………………………..……………………. ii
Acknowledgments ………………………………………………………………….... iii
Table of Contents ……………………………………………………………………….. iv
List of Figure………………………………………………………………………….... vi

I. Introductions
1. Skin Microbiome……………………………………………………................. 1.
1.1. Trichophyton rubrum…………………………………………………………… 1.
1.2. Staphylococcus epidermidis………………………………….................... 1.
1.3. Staphylococcus lugdunensis ……………………………………………... 2.
1.4. Propionibacterium acnes……………………………………………………. 3.
1.5. Escherichia coli …………………………………………………………. 4.
2. Interaction of Skin Micro Flora………………………………........................... 4.
2.1. Bacterium fungi interaction…………………………………………….... 4.
2.2. Bacterium – bacterium interaction……………………………………...... 5.
3. Fermentation of Bacteria……………………………………………………….. 6.
4. Monoclonal Antibody………………………………………………………….. 7.
5. mPEG-PCL synthesis………………………………………………………..… 8.

II. Materials
1. Apparatus or instrument………………………………………………………... 9.
2. Reagent ………………………………………………………………………... 9.

III. Methods
1.1. Growth of bacteria……………………………………….…………………… 11.
1.2. Fermentation experiment……………………………………………………... 11.
2.1. Preparation of mPEG – PCL……………………………………………….. 11.
2.2. Spectrophotometer measurement principal………………………………… 12.
2.3. Phenol Red acid – base indicator experiment………………………………. 12.
2.4. Method……………………………………………………………………... 12.
3. Two layer assay………………………………………………………………...….... 13.
4. In vitro neutralization assay………………………………………………………… 13.
5. In vivo neutralization assay…………………………………………………………. 14.
6. Fermentation of bacteria in vivo……………………………………………………. 15.
7. ELISA………………………………………………………………………………. 16.

IV. Result
1. Interference of Commensal S.epidermidis with the Growth of T.rubrum in the presence of Glycerol……………………………………………………………………………. 17.
2. Bacteria fermentation in vitro………………………………………………………… 17.
3. S.epidermidis Sucrose Fermentation Abrogated P.acnes – Induced Inflammation and Bacteria Colonization in vivo……………………………………………………… 18.
4. Using Monoclonal Antibody against P.acnes lipase in vivo……………….... 18.
V. Discussions…………………………………………………………………………. 17.
VI. References………………………………………………………………………...… 22.
VII. Appendixes ………………………………………………………………………… 29.

參考文獻

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指導教授

黃俊銘(Huang Chun Ming)

審核日期

2017-1-24

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