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研究生:曾文楷
研究生(外文):Wen-kai Tseng
論文名稱:十種中草藥之安全性及化粧品效能的研究
論文名稱(外文):Study of Ten Chinese Herbs on the Safety and Cosmetic Efficacy
指導教授:官常慶官常慶引用關係
指導教授(外文):Chang-Chin Kwan
學位類別:博士
校院名稱:靜宜大學
系所名稱:應用化學研究所
學門:自然科學學門
學類:化學學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:201
中文關鍵詞:痤瘡溶血突變性中草藥黑色素抗氧化紫外光
外文關鍵詞:mutagenicityChinese herbshemolysismelaninacneantioxidationultraviolet radiation
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本研究針對皂角刺、海桐皮、菊花、白鮮皮、蛇床子、台灣天胡荽、藿香、丁香、狗尾蟲、魚針草等十種中草藥乙醇萃取物,以初步篩選的方式評估其安全性以及各項可應用於化粧品的效能性,以期開發出天然、安全、有效之化粧品原料。
本研究於安全性測試方面,採用沙門氏菌回復突變性測試法以及溶血性試驗分別探討藥品是否具有致突變性以及具眼睛刺激的可能性,結果顯示十種中草藥並無致基因突變的作用,而不具眼睛刺激性的可能性很高。
在化粧品效能方面,本研究分別測試中草藥的抗致突變性、對痤瘡病原菌的抑制能力、對黑色素生成的影響、抗氧化的能力,以及對紫外光的吸收程度。在抗致突變性方面,效果最好的為皂角刺,並兼具高度抑制框構移位和鹽基置換型的突變作用。對痤瘡病原菌的抑制能力方面,結果顯示效果最好的為丁香,兼具抑制四株痤瘡病原菌的生長而且有中-高活性抑菌性。對黑色素生成的影響方面,菊花和丁香有較佳抑制Dopachrome生成的效果,IC50分別為是0.76 mg/ml和0.94 mg/ml。在抗氧化性方面,菊花兼具良好的四種抗氧化機制,皂角刺、藿香和丁香則分別在清除DPPH˙、還原力、清除O2˙-方面都有明顯的表現。當測定對紫外光的吸收程度時,結果顯示皂角刺、蛇床子在UV-A波段下有良好的吸收效果;蛇床子在UV-B波段下也有良好的吸收作用。
In this study, we test the safety of ethanolic extracts of Gleditsia sinensis Lam., Erythrina variegata L., Chrysanthemum morifolium Ramat., Dictamnus dasycarpus Turcz., Cnidium monnieri (L.) Cuss., Hydrocotyle formosana Masamune, Pogostemon cablin (Blanco) Benth., Eugenia caryophyllata Thunb., Heliotropium indicum L., Anisomeles indica O. Kuntze and estimate their cosmetic efficacy by using several kinds of experiments in order to primarily screen these Chinese herbs to gain natural, safe, and effective cosmetic ingredients.
Salmonella/microsome reversion assay and hemolysis test have been adopted as safe tests of these ten Chinese herbs. Using these two methods we intend to evaluate if Chinese herbs cause mutation and stimulate eyes. Results reveal that there are non-mutagenic functions and high possibility of not stimulating eyes from these ten Chinese herbs.
In the aspect of cosmetic efficacy, these extracts of Chinese herbs are used to test antimutagenicity, the ability of inhibiting acne pathogens, the effects on the formation of melanin, the ability of antioxidation and the state of absorbing ultraviolet light. We found that, about antimutagenicity, Gleditsia sinensis Lam. possesses strong activity of inhibiting frame-shift mutation and base-pair substitution mutation. About inhibiting acne pathogens, the results show that Eugenia caryophyllata Thunb. possesses medium to high activity of inhibiting four different acne pathogens. About effecting on the formation of melanin, Chrysanthemum morifolium Ramat. and Eugenia caryophyllata Thunb. possess significant efficiency of inhibiting the formation of melanin, with IC50 0.76mg/ml and 0.94 mg/ml respectively. About antioxidative effects, Chrysanthemum morifolium Ramat. demonstrates all four different antioxidative mechanisms. Gleditsia sinensis Lam., Pogostemon cablin (Blanco) Benth. and Eugenia caryophyllata Thunb. show good abilities of scavenging DPPH˙, reducing power and scavenging O2˙-. When extracts of Chinese herbs are used to test the absorption of UV light, the result shows that Gleditsia sinensis Lam. possesses good absorptive efficiency under UV-A wavelength. and Cnidium monnieri (L.) Cuss. possesses obvious absorptive efficiency under UV-A and UV-B wavelength.
中文摘要……………………………………………………………Ⅰ
英文摘要……………………………………………………………Ⅲ
總目錄………………………………………………………………Ⅴ
目錄……………………………………………………….………..Ⅵ
圖表索引…………………………………………………………...Ⅹ

目 錄
壹、緒論……………………………………………………………..1
1-1前言………………………………………………………………1
1-2皮膚的構造…………………………………………………………….3
1-3皮膚與污染原………………………………………………………….4
1-4沙門氏菌回復突變性測試法……………………………………….....5
1-4-1背景……………………………………………………………..5
1-4-2沙門氏菌的特徵………………………………………………..6
1-4-3沙門氏菌組胺酸生合成途徑…………………………………..6
1-4-4沙門氏菌組胺酸操縱組模式…………………………………..8
1-4-5致突變性的評估………………………………………………..8
1-4-6抗致突變的評估………………………………………………11
1-4-7突變的分類……………………………………………….…...11
1-4-8突變導致的結果………………………………………………12
1-4-9突變的成因……………………………………………………13
1-5溶血性試驗…………………………………………………………...17
1-5-1眼睛刺激性試驗………………………………………….…...18
1-5-2眼睛刺激性試驗以溶血性試驗取代………………….….…..19
1-5-3血紅素變性試驗…………………….….……………………..20
1-6痤瘡的簡介…………………………………………………………...21
1-6-1皮脂腺的介紹………………………………………………....21
1-6-2痤瘡的成因……………………………………………………21
1-6-3痤瘡的病灶及發展……………………………………….…...24
1-6-4痤瘡的治療藥物………………………………………….…...26
1-6-5痤瘡病原菌及其它共生菌的特徵………………………...….28
1-7與黑色素相關的背景資料……………………………………….…..30
1-7-1黑色素細胞……………………………………………………30
1-7-2酪胺酸酶的簡介…………………………………………..…..30
1-7-3膚色與黑色素……………………………………………..…..32
1-7-4黑色素的生成路徑……………………………………….…...33
1-7-5黑色素細胞的分化及黑色素在皮膚的移行途徑……….…...34
1-7-6黑色素生成的抑制作用………………………………….…...35
1-7-7黑色素生成的促進作用………………………………….…...39
1-8皮膚的老化………………………………….….…………………….43
1-8-1皮膚老化的內在因素……………….….……………………..43
1-8-2皮膚老化的外在因素……………….….……………………..44
1-8-3氧化壓力………………………………………………………46
1-8-4抗氧化防禦系統………………………………………………50
1-9紫外光與防曬…………………………………………………….…..53
1-9-1紫外線對皮膚的影響………………………………….……...53
1-9-2防曬劑的分類…………………………………………………55
1-10試驗用中草藥之簡介……………………………………………….58
貳、實驗部分……………………………………………………………..82
2-1實驗材料……………………………………………………………...82
2-2實驗方法………………………………………………………….…..87
2-2-1中草藥萃取物製備方法………………………………………87
2-2-2沙門氏菌回復突變性測試法…………………………………87
2-2-3溶血性試驗……………………………………………………97
2-2-4痤瘡病原菌抑制試驗………………………………………..100
2-2-5對黑色素生成的影響………………………………………..105
2-2-6抗氧化能力的測定…………………………………………..107
2-2-7對於紫外光吸收的測定……………………………………..114
參、結果….…….….…………………….…….…….……………116
3-1中草藥萃取率……………………………………………………….116
3-2沙門氏菌回復突變性測試法……………………………………….116
3-3溶血性試驗………………………………………………………….118
3-4痤瘡病原菌抑制試驗……………………………………………….119
3-5對黑色素生成的影響……………………………………………….122
3-6抗氧化能力的測定………………………………………………….122
3-7對於紫外光吸收的測定…………………………………………….124
肆、討論….…….….………………..….…….….…………….…169
4-1中草藥萃取率.…………………………………….……………169
4-2沙門氏菌回復突變性測試法……………………………………169
4-3溶血性試驗.…………………………………………………….171
4-4痤瘡病原菌抑制試驗…………………………………………...171
4-5對黑色素生成的影響…………………………………………...172
4-6抗氧化能力的測定……………………………………………...173
4-7對於紫外光吸收的測定…………………………………………175
伍、結論.…….….………………..….….….………………….…176
陸、參考文獻.….………………..….…….……………………..179


圖表索引
圖1- 1皮膚的結構.…………………………….……………………71
圖1- 2沙門氏菌組胺酸生合成途徑……………….…………………71
圖1- 3沙門氏菌組胺酸操縱組模式……………….…………………72
圖1- 4導致痤瘡的因子.………………….…………………….……72
圖1- 5黑色素細胞.…………………………………….……….……73
圖1- 6酪胺酸酵素的活化.…………………………………………..73
圖1- 7酪胺酸酵素的功能.………………………………………......74
圖1- 8黑色素的生成路徑.……………………………………...…..74
圖1- 9黑色素細胞的生成與黑色素體新陳代謝的過程.……..………75
圖1- 10表皮黑色素單位.……………………………………...…….76
圖1- 11 Hydroquinone氧化競爭酪胺酸酵素的機制.………….……..76
圖1- 12 Methimazole-O-Quinone之共軛結構.……………………….77
圖1- 13活性氧的生成以及抗氧化劑的作用.………………………..78
圖3- 1中草藥之萃取率.……………….…………………………...126
圖3- 2血紅素的紫外光-可見光吸收圖譜.……………….…………148
圖3- 3中草藥萃取物之log HC50.……………….………………….151
圖3-41.0mg/ml中草藥萃取物對Dopachrome 生成的抑制率………156
圖3-5100.0mg/ml中草藥萃取物對DPPH自由基的清除率…………158
圖3-6不同濃度中草藥萃取物對DPPH自由基的清除率………….160
圖3-7不同濃度中草藥萃取物對鐵離子的還原力………………….162
圖3-8100.0mg/ml中草藥萃取物對亞鐵離子的螯合能力…….........163
圖3-9不同濃度中草藥萃取物對亞鐵離子的螯合能力……………..164
圖3-101.0mg/ml中草藥萃取物對超氧陰離子的清除率…………....165
圖3-11不同濃度中草藥萃取物對超氧陰離子的清除率……………166
表1- 1眼睛損傷嚴重性的評分表…………………………………….79
表1- 2歐盟各國禁止已有取代方法的動物試驗時間表……………...80
表1- 3歐盟各國禁止沒有取代方法的動物試驗時間表……………...81
表3- 1沙門氏菌(Salmonella typhimurium)試驗菌株基因型…………127
表(3-2)-1~表(3-11)-1不同濃度中草藥萃取物對沙門氏菌之毒性試驗…………………………………………………….128~147
表(3-2)-2~表(3-11)-2不同濃度中草藥萃取物對沙門氏菌之致突變性試驗……………………………………………………..128~147
表(3-2)-3~表(3-11)-3不同濃度中草藥對沙門氏菌之抗致突變性試驗. ……………………………………………………128~147
表3-12中草藥萃取物之血紅素變性率……………………………..149
表3-13中草藥萃取物之logHC50……………………………………150
表3-14中草藥萃取物對Propionibacterium acnes之抑菌活性………152
表3-15經由篩選後之中草藥萃取物對Propionibacterium acne之最小抑菌濃度(MIC) ……………………………………………...........152
表3-16中草藥萃取物對Staphylococcus aureus subsp. aureus之抑菌活性………………………………………………………………..153
表3-17經由篩選後之中草藥萃取物對Staphylococcus aureus subsp. aureus之最小抑菌濃度(MIC) …………………………………153
表3-18中草藥萃取物對Staphylococcus epidermidis之抑菌活性……154
表3-19經由篩選後之中草藥萃取物對Staphylococcus epidermidis之最小抑菌濃度(MIC) ……………………………………………...154
表3-20中草藥萃取物對Malassezia furfur之抑菌活性………………155
表3-21經由篩選後之中草藥萃取物對Malassezia furfur之最小抑菌濃度(MIC) ………………………………………………………...155
表3-22中草藥萃取物對Dopachrome 生成的IC50……………………157
表3-23不同濃度中草藥萃取物對DPPH自由基的清除率…………..159
表3-24不同濃度中草藥萃取物對鐵離子的還原力…………………..161
表3-25不同濃度中草藥萃取物對亞鐵離子的螯合能力……………..164
表3-26不同濃度中草藥萃取物對超氧陰離子的清除率……………..166
表3-27中草藥萃取物於UV-A~UV-B範圍之最大吸收波長.……......167
表3-28中草藥萃取物於UV-A、UV-B之吸光值積分面積.…………168
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