臺灣博碩士論文加值系統

Propionibacterium acne、Staphylococcus aureus與Staphylococcus epidermidis為痤瘡病灶中之常見菌株，本研究係針對六十種中藥材之熱水萃出物，進行此三株菌株之抑菌性試驗。由disc diffusion assay 篩選出五倍子、黃芩與黃柏對於P. acne BCRC10723、S. aureus BCRC10451與S. epidermidis BCRC10823有顯著之抑菌效果。五倍子、黃芩與黃柏之熱水萃出物對 P. acne BCRC10723之最小抑菌濃度(minimum inhibitory concentration, MIC)分別為0.3125 mg/ml、0.3125 mg/ml與0.625 mg/ml；對S. aureus BCRC10451之MIC分別為0.625 mg/ml、0.625 mg/ml與2.5 mg/ml；對S. epidermidis BCRC10783之MIC則分別為 0.3125mg/ml、0.625 mg/ml與2.5 mg/ml。五倍子、黃芩與黃柏之熱水萃出物經由n-hexane、ethyl acetate、diethyl ether與1-butanol加以區分後，可初步分離出五倍子之抑菌成分位於ethyl acetate區分，黃芩與黃柏之抑菌成分則於1-butanol區分。

Propionibacterium acne , Staphylococcus aureus and Staphylococcus epidermidis are the common bacterial strains in the acne lesions. Preliminary screening of hot water extracts of sixty of traditional Chinese herbal medicines for antibacterial activities conducted . Rhus chinensis, Scutellaria baicalensis and Phellodendron amurense with antibacterial activitie of P. acne BCRC10723 , S. aureus BCRC10451 and S. epidermidis BCRC10823 were found by disc diffusion assay, the minimum inhibitory concentrations (MIC) of hot water extracts of R. chinensis, S. baicalensis and P. amurense for P. acne BCRC10723 were 0.3125 mg/ml , 0.3125 mg/ml and 0.625 mg/ml, respectively; For S. aureus BCRC10451, the MICs were 0.625 mg/ml , 0.625 mg/ml and 2.5 mg/ml, respectively; For S. epidermidis BCRC10783, the MICs were 0.3125mg/ml , 0.625 mg/ml and 2.5 mg/ml, respectively . After fractionation of R. chinensis, S. baicalensis and P. amurense hot water extracts with n-hexane, followed by ethyl acetate , diethyl ether and 1 – butanol, ethyl acetate fraction of R. chinensis and 1 - butanol fraction of S. baicalensis and P. amurense showed the primising antibacterial activity.

中文摘要............................I
英文摘要...........................II
壹、前言...........................1
貳、文獻整理........................3
一、皮膚的構造......................3
(一) 角質細胞......................3
(二) 黑色素細胞....................3
(三) 朗格罕氏細胞....................6
(四) 皮膚細胞的代謝..................6
二、痤瘡之病灶及發展 .................6
(一) 非發炎性痤瘡 .................. 8
1. 閉鎖性粉刺........................8
2. 開放性粉刺........................ 8
(二) 發炎性痤瘡 ..................... 8
(三) 痤瘡之併發症 ................... 10
1. 猛爆性痤瘡 ........................ 10
2. SAPHO 症候群 ........................ 11
3. PAPA 症候群 ......................... 11
三、 痤瘡之形成機制.................... 11
(一) 毛囊之過角質化 ................... 11
(二) 皮脂的過度增生..................... 12
(三) 痤瘡病源菌之滋生................... 12
1. 痤瘡桿菌 ............................ 13
2. 金黃色葡萄球菌 ....................... 15
3. 表皮葡萄球菌 ........................ 15
(四) 發炎反應............................ 16
四、痤瘡之治療方式 ...................... 16
(一) 局部外用藥劑 ....................... 16
1. 維他命A酸 ........................... 16
2. 痘膚潤 ............................... 20
3. 水楊酸 ............................... 20
4. 杜鵑花酸 ............................ 20
5. 過氧化苯 ............................ 20
6. 抗生素外用藥 ........................ 20
(1) 作用機轉............................ 20
(2) 使用方法............................ 21
(3) 副作用.............................. 21
7. 治療痤瘡外用藥的合併使用 ............ 21
(二) 全身性藥物治療 .................... 21
1. 口服抗生素 .......................... 22
2. 口服維他命A酸 ....................... 22
3. 賀爾蒙治療 .......................... 22
4. 注射類固醇 .......................... 23
(三) 物理性治療 ........................ 23
五、中草藥之開發與應用.................. 23
(一) 全球中草藥之市場趨勢及概況 ........ 23
(二) 中草藥於化妝品市場之潛力及優勢 .... 25
(三) 台灣中草藥化妝品發展之瓶頸及優勢 .. 26
六、中草藥抑制痤瘡桿菌之相關研究 ....... 28
(一) 中醫文獻記載 ...................... 29
(二) 中醫病因病機 ...................... 29
(三) 中醫的辯證論治 .................... 30
(四) 藥理研究 .......................... 30
(五) 外用治療法 ........................ 31
參、 實驗設備與方法 .................... 33
一、實驗材料與藥品...................... 33
(一) 試驗菌株........................... 33
(二) 試驗之中藥材....................... 33
(三) 試藥及儀器 ........................ 34
1. 試藥 ................................ 34
2. 儀器設備 ............................ 35
二、實驗方法............................ 35
(一) 培養液及培養基之製備 .............. 35
(二) 菌種保存與活化 .................... 37
(三) 接種源之製備 ...................... 37
(四) 菌數之測定......................... 38
(五) 中藥熱水萃取物之製備............... 38
(六) 中藥熱水萃取液之抑菌試驗 .......... 39
(七) 最小抑菌濃度試驗與最小殺菌濃度試驗...39
(八) 中藥熱水萃取物不同濃度對痤瘡病原菌之抑菌性試驗...... 40
(九) 中藥熱水萃取物MBC對痤瘡病原菌之生長曲線試驗 ....... 41
(十) DMSO效應試驗.................. 41
(十一) 有機溶劑分層區分試驗 ........41
三、統計分析 ...................... 42
肆、 結果與討論 .................... 42
一、六十種中藥熱水萃取物對痤瘡病原菌之抑菌性. 42
二、具抑菌性之中藥熱水萃取物對痤瘡病原菌之MIC與MBC......46
三、 中藥熱水萃取物不同濃度對痤瘡病原菌之抑菌性 .......... 52
四、 DMSO濃度對試驗菌株生長之影響 .......... 58
五、 有機溶劑分層區分之抑菌試驗 ........... 62
(一) 有機溶劑對熱水萃取物之分層回收率....... 62
(二) 黃柏、五倍子及黃芩熱水萃取物之各分層對痤瘡病原菌之抑菌性 .... 66
(三) 黃柏、五倍子及黃芩熱水萃取物各分層對痤瘡病原菌之MIC與MBC .... 68
伍、 結論 .............................. 73
陸、 參考文獻 ................... 75
柒、 附錄 ............................... 84

甘偉松。藥用植物學。國立中國醫藥研究所出版，台中。
江淑芬。2000。植物來源藥市場、技術及應用專題調查。工研院產業經濟與資訊服務中心出版，新竹。
李時珍。本草綱目。文化圖書公司，台北。
李文森。1997。解剖生理學。華杏出版社，台北。
沈明來。1993。試驗設計學。九州圖書公司，台北。
吳敏綺、邱琬婷、徐自菱、林文勝。2001。中西醫會診－青春痘。書泉出版社，台北。
林天樹。2000。老師傅鑑定中藥(第一~四冊)。林石金出版社，台北市。
曾文楷。2002。中草藥對於痤瘡病源菌與黑色素生成的影響。靜宜大學應用化學研究所碩士論文，台中。
周丹。1990。內外兼治痤瘡126例療效觀察。內蒙古中醫藥，9：14。
許世昌。1992。最新實用解剖生理學。永大書局，台北。
許芳華。2001。黃芩抑菌性質及抑菌成分分離純化之研究。中興大學食品科學研究所碩士論文，台中。
許堯欽。2001。青春痘的中醫療法。台南縣中醫師公會會刊，台南。
梁文俐。2002。市售五倍子品質及抗菌活性探討。中醫藥委員會研究計劃，台北。
張慈映。2003。深具發展潛力的生技蛋白質藥品市場。產業技術資訊服務推廣計畫中心出版，台北。
秦慶瑤。2001。生技及製藥產業。財團法人生物技術開發中心，台北。
巢佳莉。2001。生技事業發展成功案例分析。財團法人生物技術開發中心，台北。
楊惠雯。2003。含中藥美容製劑療效之臨床評估。中國醫藥學院藥物化學研究所碩士論文，台中。
趙昭明。2004。還我一張潔淨的臉。大樹林出版社，台北。
趙彥宇與邱慧嫻。2002。啊！青春痘完全搞定。原水文化，台北。
廖美智。2001。中草藥產業現況與趨勢。財團法人生物技術開發中心，台北。
戴新民。1986。中藥臨床應用。啟業書局，台北。
Alzoreky, N. S., and K. Nakahara. 2003. Antibacterial activity of extracts from some edible plants commonly consumed in Asia. Int. J. Food Microbiol. 80: 223-230.
Arias, M. E., J. D. Gomez, N. M. Cudmani, M. A. Vattuone, and M. I. Isla. 2004. Antibacterial activity of ethanolic and aqueous extracts of Acacia aroma Gill. Ex Hook et Arn. Life Sci. 75: 191-202.
Ahn, Y. J., C. O. Lee, J. H. Kweon, J. W. Ahn and J. H. Park. 1998. Growth-inhibitory effect of Galla Rhois-derived tannins on intestinal bacteria. J. Appl. Microbiol. 84: 439-443.
Ashkenazi, H., Z. Malik, Y. Harth and Y. Nitzan. 2003. Eradication of Propionibacterium acnes by its endogenic porphyrins after illumination with high intensity blue light. FEMS Immuno. and Med. Microbiol. 35: 17-24
Bassett, I. B., D. L. Pannowitz and R. S. Barnetson. 1990. A comparative study of tea-tree oil versus benzoylperoxide in the treatment of acne. Med. J. of Aus. 153: 455-458.
Berghe, D. A. V. and A. J. Vlietinck. 1991. Screening methods for antibacterial and antiviral agents from higher plants. In: Dey, P.M., Harborne, J.B. (Eds.), Methods in Plant Biochemistry. Academic Press, London. 6: 47- 69.
Bibel, D. J., R. J. Smiljanic, and D. J. Lovell. 1978. Interactions of Bacillus licheniformis ATCC 10716 and Normal Flora of Human Skin. Appl. and Environ. Microbiol. 35：1136-1144.
Brantner, A., Z. Males, S. Pepeljnjak and A. Antolic. 1996. Antimicrobial activity of Paliurus spina-christi mill. J. Ethnopharmacol. 52: 119-122.
Caelli, M., J. Porteous, C. F. Carson, R. Heller, and T. V. Riley. 2000. Tea tree oil as an alternative topical decolonization agent for methicillin- esistant Staphylococcus aureus. J Hospit. Infec.46:236-237.
Cammue, B. P., B. M. F. De, F. R.Terras, P. Proost, V. J. Damme, S. B. Rees, J. Vanderleyden, and W. F. Brookaert. 1992. Isolation and characterization of a novel class of plant antimicrobial peptides form Mirabilis jalapa L. seeds. J. Biol. Chem. 267:2228-33.
Cho, E. J., T. Yokozawa, D. Y. Rhyu, S. C. Kim, N. Shibahara, and J. C. Park. 2003. Study on the inhibitory effects of Korean medicinal plants and their main compounds on the 1,1-diphenyl-2-picrylhydrazyl radical. Phytomedicine. 10:544-51.
Cortis E., F. D'Benedetti, A. Insalaco, S. Cioschi, F. Muratori, L. E. D'Urbano and A. G. Ugazio. 2004. Abnormal production of tumor necrosis factor (TNF) -- alpha and clinical efficacy of the TNF inhibitor etanercept in a patient with PAPA syndrome [corrected]. J Pediatr. 145:851-855.
Cunliffe, W. J., K. T. Holland, R. Bojar and S. F. Levy. 2002. A randomized, double-bind comparison of a clindamycin phosphate/ benzyol peroxide gel formulation and a matching clindamycin gel with respect to microbiologic activity and clinical efficacy in the topical treatment of acne vulgaris. Clin. Therapy 24: 1117-1133.
Deena, M. J. and J. E. Thoppil. 2000. Antimicrobial activity of the essential oil of Lantana camara. Fitoterapia 71:453-455.
Esperson, F. 1998. Resistance to antibiotics used in dermatological practice. Br. J. Dermatol. 139:4-8.
Farrar, M. D. and E. Ingham. 2004. Acne: Inflammation. Clinics in Dermatology. 22: 380–384.
Graham G. M., M. D. Farrar and J. E. Cruse-Sawyer. 2004. Proinflammatory cytokine production by human keratinocytes stimulated with Propionibacterium acnes and P. acnes GroEL. Br. J. Dermatol. 150:421-428.
Gun, W. S. 1986. Pharmaceutical botany National Research Institute Of Chinese Medicine. Taipei, Taiwan.
Guy, R., M. Green and T. Kealey. 1996. Modeling of acne in vitro. J. Invest. Dermatol. 106: 176-182.
Harrison C. J., M. I. Marks and D. F. Welch. 1985. Microbiology of recently treated acute otitis media compared with previously untreated acute otitis media. Pediatr. Infect. Dis. 4: 641-646.
Hayes, A. J. and B. Markovic. 2002. Toxicity of Australian essential oil Backhousia citriodora (Lemon myrtle). Part 1. Antimicrobial activity and in vitro cytoxicity. Food and Toxico. 40: 535-543.
Higaki, S., S. Morimatsu, M. Morohashi, T. Yamagishi, and Y. Hasegawa. 1997. Susceptbility of Propionibacterium acnes, Staphylococcus aureus and Staphylococcus epidermidis to 10 Kampo formulations. J. Int. Med. Res. 25: 318-324.
Higaki, S. 2003. Lipase inhibitors for the treatment of acne. J. molec. Catal. B: Enzymatic. 22:377-384.
Holland, K. T., I. Eileen, and W. J. Cunlife. 1981. A review the microbiology of acne, J. Appl. Bacteriol. 51:195-215.
Hunter, J. A. A., J. A. Savin, and M. V. Dahl. 1989. Clinical Dermatology, Blackwell Scientific Publications.
Iqbal M. and M. S. Kolodney. 2005. Acne fulminans with synovitis-acne-pustulosis-hyperostosis-osteitis (SAPHO) syndrome treated with infliximab. J. Am. Acad. Dermatol. 52:118-120.
Junho, K., J. Lee, E. Jung, Y. Park, K. Kim, B. Park, K. Jung, E. Park, J. Kim and D. Park. 2004. In vitro antibacterial and anti-inflammatory effects of honokiol and magnolol against Propionibacterium sp Eur. J. of Pharma. 496: 189-195.
Kapsokefalou, M. and A. Salifoglou. 2003. Antioxidant activity of organic extracts from aqueous infusion of sage. J. Agric. Food Chem. 51: 6696-6701.
Kloos, W. and M. S. Musselwhite. 1975. Distribution and persistence of Staphylococcus and Micrococcus Species and Other Aerobic Bacteria on human skin. Appl. Microbio. 30: 381-395.
Knaggs, H., K. Holland, C. Morris, E. Wood and W. Cunliffe. 1994. Quantification of cellular proliferation in acne using the monoclonal antibody Ki-67. J. Invest. Dermatol. 102: 89-92.
Krautheim, A. and H. P. Gollnick. 2004. Acne: topical treatment. Clin. Dermatol. 22:398-407.
Kubo, I., K. Hori, Y. Kubo, Y. Yamagiwa, T. Kamikawa, H. Haraguchi. 2000. Molecular design of antibrowning agents. J. Agric. Food Chem. 48:1393-1399.
Kubo. I., Y. Xu and K. Shimizu. 2004. Antibacterial activity of ent-kaurene diterpenoids from Rabdosia rosthornii. Phytother. Res. 18: 180-183.
Kusamba, C., K. Byamana, and W. M. Mbuyi. 1991. Antibacterial activity of Mirabilis jalapa seed powder. J. Ethnopharmacol. 35:197-199.
Leyden, J. J. 2001. The development of antibiotic resistance in Propionibacterium acnes. Cutis. 67: 21-24.
Matsingou T. C., N. Petrakis, M. Kapsokefalou and A. Salifoglou. 2003. Antioxidant activity of organic extracts from aqueous infusions of sage. J. Agric. Food Chem. 51:6696-701.
Mehrany K, J. M. Kist, R. H. Weenig and P. M. Witman. 2005. Acne fulminans. Int. J. Dermatol. 44:132-133.
Okeke, M. I., C. U. Iroegbu, E. N. Eze, A. S. Okoli, and C. O. Esimone. 2001. Evaluation of extracts of the root of Landolphia owerrience for antibacterial activity. J. Ethnopharmaco. 78: 119-127.
Orafidiya, L. O., E. O. Agbani, A. O. Oyedele, O. O. Babalola and O. Onayemi. 2002. Preliminary clinical tests on topical preparations of Ocimum gratissimum Linn leaf essential oil for the treatment of acne vulgaris. Clinical Drug Investigations 22: 313-319.
R�縵nier, M., M. J. Staquet, D. Schmit, and R. Schmidt. Integeation of langerhans cells into a pigmented reconstructed human epidermis. 1997. J. Invest. Dermatol.. 109:510-512.
Ross, J. I., A. M. Snelling, E. A. Eady, J. H. Cove, W. J. Cunliffe, J. J. Leyden, P. Collignon, B. Dreno, A. Reynaud, J. Fluhr, and S. Oshima. 2001. Phenotypic and genotypic characterization of antibiotic-resistant Propionibacterium acnes isolated from acne patients attending dermatology clinics in Europe, the USA, Japan, and Australia. British J. Dermatol. 144: 339-346.
Russell J. J. 2000. Topical therapy for acne. America. Family Physic. 61: 357-365.
Saha, K., N. H. Lajis, A. S. Hamzah, S. Khozirah, S. Khamis and A. Syahida. 2004. Evaluation of antioxidant and nitric inhibitory activities of selected Malaysian medicinal plants. J. Ethnopharmaco. 92:263-267.
Sheu, S. C. 2002. The newest practical physiological anatomy, Yung-Da, Taipei, Taiwan
Shuichi, H. 2003. Lipase inhibitors for the treatment of acne. J. Molecular Catalysis B: Enzymatic, 22: 377-384.
Swanson, j. K. 2003. Antibiotic resostance of Propionibacterium acnes in acne vulgaris. Dermatol. Nurs. 15: 359-362.
Thiboutot D., G. Harris, V. Iles, G. Cimis, K. Gilliland and S. Hagari. 1995. Activity of the type 1 5α-reductase exhibits regional differences in isolated sebaceous glands and whole skin. J. Invest. Dermatol. 105:209-214.
Thiboutot D., H Knaggs, K. Gilliland and S. Hagari. 1997. Activity of the type 1 5α-reductase is greater in the follicular infrainfundibulum comparedto the epidermis. Br. J. Dermatol. 136:166-171.
Thiboutot, D. 2002. Acne: 1991-2001. J. Am. Acad. Dermatol.
47:109-117.
Usatine, R. P. and M. A. Quan. 2000. Pearls in the management of acne. Primary Care: Clinic. Office Prac. 27:289-308.
Warner G. T. and Plosker G. L. 2003. Clidamycin/benzoyl peroxide gel: a review of its use in the management of acne. Am. J. Clin. Dermatol. 3: 349-360.
Webster, G. F. 2002. Acne vulgaris. Br. Med. J. 325:475-479.
White, S. D., and J. A. Yager. 1995. Resident dendritic cells in the epidermis: langerhans cells markel cells and melanocytes. Veterinary Dermatol. 6:1-8.
Yang D., H. Hu, S. Huang, J.P. Chaumont and J. Millet. 2000. Study on the inhibitory activity, in vitro, of baicalein and baicalin against skin fungi and bacteria. Zhong Yao Cai. 23: 272-274.