臺灣博碩士論文加值系統

夏枯草(Prunella vulgaris L.)是屬於唇形科的植物，文獻記載其具有清肝、解毒、降血等功效及對大腸桿菌、葡萄球菌等具有抑制作用的藥理作用。
在我們持續致力於化妝品天然原料的開發研究中，最近發現夏枯草之萃取物具有使酪胺酸酶(tyrosinase)活性增強的效果。
本研究以tyrosinase活性導向與管柱層析得到其活性成分，並經由光譜分析確定其結構。

In our continuing search for natural source ingredient from Chinese medicinal herbs, was found that extracts of Prunella vulgaris L. showed the significant tyrosinase enhancing activity.
In this study, the active constituent of Prunella vulgaris L. was isolated by the process of tyrosinase activity assay and column chromatography. The structure was elucdiate by the spectroscopic methods and its activity was also achieved.

中文摘要---------------------------------------------------------------1
英文摘要---------------------------------------------------------------2
壹. 前言---------------------------------------------------------------3
貳. 植物介紹-----------------------------------------------------------4
參. 活性測試原理-------------------------------------------------------9
一. 皮膚之結構-------------------------------------------------------9
1. 表皮(Epidermis)-------------------------------------------------9
2. 真皮(Dermis)---------------------------------------------------10
3. 皮下組織(Subcutis)---------------------------------------------11
二. 黑色素(Melanin)之產生過程-----------------------------------------13
1. 黑色素細胞(Melanocyte)-----------------------------------------13
2. 黑色素(Melanin)------------------------------------------------15
(1) 真黑色素(Eumelanin)------------------------------------------15
(2) 褐黑色素(Pheomelanin)----------------------------------------15
3. 黑色素的功用--------------------------------------------------16
(1) 吸收紫外線---------------------------------------------------16
(2) 穩定自由基---------------------------------------------------16
(3) 可做為生物聚合物(Biopolymer)---------------------------------16
4. 黑色素生成(Melanogensis)的機轉---------------------------------17
(1) 黑色素之生化合成路徑-----------------------------------------17
(2) 黑色素細胞中之黑色素生成過程---------------------------------18
三. 酪胺酸酶與黑色素之關係------------------------------------------21
1. 酪胺酸酶(Tyrosinase)-------------------------------------------21
2. 影響酪胺酸酶活性的因素-----------------------------------------21
(1) 內分泌的影響-------------------------------------------------21
(2) 化學方面的影響-----------------------------------------------22
(3) 物理因素的影響-----------------------------------------------23
四. 促進酪胺酸酶活性的物質及方法------------------------------------24
1. 增加酪胺酸酶的合成---------------------------------------------24
2. 直接增加黑色素的生成-------------------------------------------24
3. 補骨脂素-------------------------------------------------------24
4. 解除對酪胺酸酶的抑制-------------------------------------------25
5. 刺激酪胺酸酶的活性---------------------------------------------25
6. 可曬黑防曬劑---------------------------------------------------26
五. 紫外線與皮膚的關係----------------------------------------------28
肆. 實驗部份----------------------------------------------------------31
一. 儀器------------------------------------------------------------31
二. 藥品------------------------------------------------------------32
三.植物之萃取及初測試-------------------------------------------------33
四. 夏枯草之萃取與成分分離------------------------------------------34
五. 試藥的配製------------------------------------------------------38
六. Tyrosinase activity之測定方法-----------------------------------39
伍. 結果與討論--------------------------------------------------------41
一. Tyrosinase activity之測定及結果---------------------------------41
二. 夏枯草水層之EA萃取物對Tyrosinase activity之百分率---------------47
三. 夏枯草水煮之各分離層的萃取率------------------------------------48
陸. 結論--------------------------------------------------------------49
柒. 參考文獻----------------------------------------------------------50
捌. 圖表索引----------------------------------------------------------59

1.T. J. Andreadis, US Paten 2 949 403, 16 August,1960

2.甘偉松；藥用植物學，國立中國醫藥研究所出版，台北，1981，pp485

3.顏焜熒：原色生藥學，南天書局，臺北 1983，pp100

4.張光雄、邱年永：原色台灣藥用植物圖鑑(1)，南天書局，臺北 1980，pp186

5.邱德文、吳家榮、夏同珩：本草綱目彩色藥圖(上卷)，薪傳出版社，臺北市 2001，pp231

6.楊濟華：部分中草藥美白與防曬成份研究，靜宜大學應用化學研究所碩士論文，1996

7.方明發；化妝品製造學，人光出版社，台北，1985

8.周德成；解剖生理學，昭人出版社，台北 1989，pp82-89

9.周燕輝；皮膚病，正中書局，台北 1987，pp1-10

10.光井武夫；新化妝品學，南山堂，台北，1992，pp18

11.劉承煌：皮膚病理學，中國醫藥科技出版社，北京 1991，pp74-98

12.L. A. Golgsmith：Biochemistry and physiology of the skin. Oxford University Press, Inc. New York, 1993

13.許元昱、李旺祚、郭文勵；組織學，合記圖書出版社，台北，1992

14.G. Prota：Recent advances in chemistry of melanogensis in mammals. J. Invest. Dermatol. 1980, 75, 122-127

15.J. M. Pawelek；A. M. Korner：The biosynthesis of mammalian melanin. American Scientist 1982, 70, 136-145

16.黃泰卿：中藥對酪胺酸酵素活性的影響，靜宜大學應用化學研究所碩士論文，1995

17.黃效民：黑色素之功能，食品工業月刊(四月號) 1996, pp8-12

18.J. M. Orten；O. W. Neuhaus：Human biochemistry, London, 1975

19.A. C. Guyton：Human physiology and mechanisems of disease, W. B. Saunders, London, 1987

20.C. B. Keith：Lawrence M：Melanins：Hair dyes for the future. Cosmetics ＆ Toiletries 1994, 109, 59-64

21.S. Shirota；K. Miyazaki；R. Aiyama；M. Ichioka；T. Y. Kura：Tyrosinase inhibitors from cude drugs. Biol. Pharm. Bull. 1994, 17(2), 266-269

22.A. Cantarow；B. Schepartz：Biochemistry, W. B. Saunders, London, 1963

23.R. H. Abeles；P. A. Frey；W. P. Jencks：Biochemistry, Jones and Bartlett, London, 1991

24.蘇倩雯：金銀花之促進酪胺酸酶活性成分研究，靜宜大學應用化學研究所碩士論文，2001

25.葉美嬌：蝦酪胺酸酵素抑制劑之研究，國立臺灣海洋大學水產食品科學研究所碩士論文，1994

26.G. Wong；J. Pawelek：Melanocyte-stimulating hormone promotes activation of pre-existing tyrosinase molecules in cloudman S-91 melanoma cells. Nature. 1975, 255, 644-646

27.M. K. Jimenez；W. L. Kameyama；Y. Maloy；Tomita；V. J. Hearing：Mammalian tyrosinase：Biosynthesis, processing and modulation by melanocyte-stimulating hormine. Proc. Natl. Acad. Sci. USA. 1988, 85, 3830-3834

28.B. B. Fuller；J. B. Lunsford；D. S. Iman：?Melanocyte-stimulating hormone regulation of tyrosinase in cloudman S-91 melanoma cell cultures. J. Biol. Chem. 1987, 262, 4024-4033

29.B. B. Fuller；D. H. Viskochil：The role of RNA and protein synthesis in mediating the action of MSH on mouse melanoma cells. Life Sci. 1979, 24, 2405-2416

30.P. L. Altmeyer；Stohr；H. Holzmann：Seasonal rhythm of the plasma level of ?melanocyte stimulating hormone. J. Invest. Dermatol. 1986, 86, 454-456

31.A., J. Sanchez-Ferrer；N., Rodriguez-Lopez；F. Garcia-Carmona：Tyrosinase：a comprehensive review of its mechanism. Biochim. Biophys. Acta. 1995, 1247, 1-11

32.J. R. Jara；F. Solano；J. C. Garcia-Borron；P. Aroca；J. A. Lozano：Regulation of mammalian melanogensis. The role of metal cations. Biochimica et Biophysica Acta. 1990, 1035, 276-285

33.D. C. Liebler：The role of metabolism in the antioxidant function of vitamin E. Critical Review in Toxicology 1993, 23, 147-169

34.洪偉章、李金枝、陳榮秀：化妝品原料及功能，藝軒，臺北，2000，pp272-273

35.黃美勳：中藥對酪胺酸酵素氧化作用之活性研究，中國醫藥學院碩士論文，1996

36.I. M. Valente；M. J. Minski；P. J. Peterson：Neutron activation analysis of noble metals in vegetation. J. Radioanal. Chem. 1982, 71(1-2), 115-127

37.T. Okuda；T. Hatano；I. Agata；S. Nishibe：The components of tannic activities in Labiatae plants. I. rosmarinic acid from Labiatae plants in Japan. Yakugaku Zasshi 1986, 106(12), 1108-1111

38.K. Sasaki；F. Yoshizaki：Nobiletin as a tyrosinase inhibitor from the peel of Cityus fruit. Biol. Pharm. Bull. 2002, 25(6), 806-808

39.L. Z. Piao；H. R. Park；Y. K. Park；S. K. Lee；J. H. Park；M. K. Park：Mushroom tyrosinase inhibition activity of some chromones. Chem. Pharm. Bull. 2002, 50(3), 309-311

40.K. Isao；K. H. Ikuyo：Flavonols from Saffron flower：tyrosinase inhibitory activity and inhibition mechanism. J. Agric. Food Chem. 1999, 47, 4121-4125

41.K. Isao；K. H. Ikuyo：Tyrosinase inhibitory activity of the olive oil flavor compounds. J. Agric. Food Chem. 1999, 47, 4574-4578

42.J. K. No；D. Y. Soung；Y. J. Kim；K. H. Shim；Y. S. Jun；S. H. Rhee；T. Yokozawa；H. Y. Chung：Inhibition of tyrosinase by green tea components. Life Sci. 1999, 65(21), PL241-246

43.K. Isao；K. H. Ikuyo；K. C. Swapan；K. Yumi；S. Yolanda；O. Tetsuya：Flavonols from heterotheca inuloides：tyrosinase inhibitory activity and structural criteria. Bioorganic＆Medicinal Chemistry 2000, 8, 1749-1755

44.K. Ikehata；J. A. Nicell：Characterization of tyrosinase for the treatment of aqueous phenols. Bioresource Technology 2000, 74, 191-199

45.H. S. Lee：Tyrosinase inhibitors of pulsatilla cernua root-derived materials. J. Agric. Food Chem. 2002, 50, 1400-1403

46.M. Duggan；J. Wilmott；A. Znalden：Tyrosinase the enzyme behind the tan. Cosmetics & Toiletries 1987(March), 102, 97-101

47.M. A. Pathak：Sunscreens-progress and perspectives on photorotection of human skin against UVB and UVA radiation. The Journal of Dermatology 1996, 23, 783-800

48.S. H. Pomerantz；I. G. Ances：Tyrosinase activity in human skin. Influence of race and age in newborns. J. Clin. Invest. 1975, 55, 1127-1131

49.M. D. Mitsuru；B. S. Todd；D. D. S. Shoko；M. D. Everett；B. B. Fuller：The relationship between tyrosinase activity and skin color in human foreskins. J. Invest. Dermatol. 1990, 95, 9-15

50.A. B. Lerner；T. B. Fitzpatrick：Biochemistry of melanin formation. Physiol. Rev. 1950, 30, 91-126

51.K. T. Ram；C. D. Chintamani；R. Abburi：pH-dependent interconversion of forms of tyrosinase in human skin. Biochem. J. 1988, 252, 481-487

52.R. Jermann；Dr. T. Schreier：L-erythrulose－A promising self-tanning molecule for a new generation in sunless tanning. Proceedings of the 20TH IFSCC Congress

53.K. T. Lee；J. H. Kim：Brazilin as new sunless tanning agent. 3RD ASCS Conference Taipei, Taiwan 1997, 33-36

54.K. T. Lee；J. H. Kim：Inhibitory effects of Ramulus Mori extracts on melanogenesis. Cosmetics & Toiletics 1998, 113, 65-70

55.A. Garcia；J. E. Fulton：The combination of glycolic acid and hydroquinone or kojic acid for the treatment of melasma and related conditions. Dermatol Surg 1996, 22, 443-447

56.P. Geary：Burns related to the use of psoralens as a tanning agent. Burns 1996, 22(8), 636-637

57.E. N. Marieb：Human automy and physiology. The Benjamin/Gummings Publishing Company, California, 1989