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研究生:卓奕璿
研究生(外文):Yi-Shiuan Juo
論文名稱:天然細胞保護劑-Ectoine於化妝品產業之應用與評估
論文名稱(外文):Evaluation and applications of a natural cell protectant-ectoine in the cosmetic industries
指導教授:魏毓宏
學位類別:碩士
校院名稱:元智大學
系所名稱:生物科技與工程研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2010
畢業學年度:98
語文別:中文
論文頁數:54
中文關鍵詞:化妝品
外文關鍵詞:EctoineMarinococcus sp.
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Ectoine為許多嗜鹽菌或耐鹽菌在高鹽環境下,為維持及平衡細胞內外滲透壓所合成的一種 compatible solute。本研究主要目的,擬將探討 Ectoine於化妝品產業之應用與評估。在本研究中,利用細胞培養、基因表現等延伸應用到老鼠皮膚等方式,初步評估 Ectoine是否具有美白,抗氧化,抗紫外線輻射等化妝品應用之潛能;並以老鼠黑色素瘤細胞(B16F0)作為美白應用測試平台。
Ectoine在美白應用效能評估方面,研究顯示:Ectoine濃度的提高可抑制黑色素生成。在64 mM濃度之 Ectoine處理下,黑色素含量明顯降低至40%,比較起 Ascorbic acid在64 mM時細胞極速死亡,更能維持其生長。在發炎反應效能評估方面,研究證實:在 UVA誘導下,Ascorbic acid 在濃度64 mM時,TNF-α含量從約114 pg/ml增加至約180 pg/ml,相反的在 64 mM 的 Ectoine,明顯抑制 TNF-α含量,降低至約30 pg/ml,而另外 UVB 刺激後,TNF-α含量也從約118 pg/ml降低至約 31 pg/ml。另外在基因分析層面探討抗氧化效能評估,Ectoine不管在有無 UV刺激下,皆可抑制 GPx1的活化,而對照組 Ascorbic acid卻在 UVA刺激下,在濃度4 mM以上反而導致 GPx1基因表現增加;保濕效能評估方面顯示,Ascorbic acid並不具有增加水合酵素的表現,而 Ectoine則在4 mM下,仍可增加水合酵素基因的表現量;在老鼠皮膚測試,經由市售之水測儀探針測得下之結果顯示,皮膚含水量可增加5~6 %,證實 Ectoine具有保濕功能。最後在防曬效能評估方面,顯示老鼠皮膚經 UVB刺激下,5 % Ectoine之處理明顯減少紅腫的發炎情形。由上述研究結果顯示,Ectoine是天然的細胞保護劑,亦同時具有美白、抗氧化、保濕、防曬等功能,證實 Ectoine具備有全方位化妝品應用之開發潛力。


Ectoine, the compatible solute that was first discovered in Ectothiorhodospira halochloris, is one of the most commonly found osmolytes in nature. The study aims to evaluate the cosmetics industrial applications for ectoine. In this study, we used cell culture and gene expression and extended the experiments on mouse skin to have a preliminary assessment of whether ectoine was a potential whitening, anti-oxidant, and anti-ultraviolet radiation agent. The animal cell used in this study was mouse melanoma cells (B16F0). The results showed that ectoine performed ability in the whitening effect. Improvement of ectoine concentration could inhibit the formation of melanin. In 64 mM of ectoine, the melanin content has been reduced to 40% when compared with 64 mM of ascorbic acid which cause the cell death to maintain its growth. In the assessment of inflammatory reaction, UVA was applied to induce cell damage. Under the treatment of 64 mM of ascorbic acid, TNF-α levels increased from about 114 pg/ml to about 180pg/ml. In the opposite of the effect of ascorbic acid, 64 mM of ectoine significantly inhibited TNF-α levels, decreased to about 30 pg/ml while TNF-α content was reduced from about 118 pg/ml to about 31 pg/ml under the stimulation of UVB. In the genetic analysis of the level of antioxidant effectiveness evaluation, regardless of whether the UV levels were, ectoine inhibited the activation of GPx1 gene expression. However, in the UVA ascorbic acid stimulation, the concentration of 4 mM led an increase in GPx1 gene expression. The results of moisture evaluation demonstrated that ascorbic acid does not increase the performance of enzyme hydration, while ectoine at 4 mM, the enzyme gene was able to enhance the expression of hydration. In mouse skin test, the results which were measure by the commercial water tester probe revealed that the skin moisture improved to 5~6%. This confirmed ectoine with moisturizing function. Finally, in mouse skin with UVB stimulation, ectoine concentrations from 1~5% of the photos showed that 5% of the ectoine decreased swelling and inflammation. The above results implied that ectoine is a natural cell protective agent, also has whitening, anti-oxidant, moisture, and even sunscreen function.

摘要 i
Abstract ii
致謝 iv
目錄 v
表目錄 viii
圖目錄 ix
第一章 緒論 1
1.1 前言 1
1.2 Ectoine簡介 1
1.3 皮膚構造 2
1.3.1 表皮組織(Epidermis) 2
1.3.2 真皮組織(Dermis) 3
1.3.3 皮下組織(Subcutaneous layer) 3
1.4 美白作用 4
1.4.1 黑色素細胞 (Melanocyte) 4
1.4.2 黑色素的生成 5
1.4.3 維生素C 5
1.4.4 抑制黑色素形成的機轉 6
1.5 防UV之曝曬效用 6
1.5.1 紫外線之介紹 6
1.5.2 紫外線分類 6
1.5.3 紫外光的反應與色素的沉澱 7
1.5.4 紫外線對皮膚的傷害 7
1.5.5 紫外線引發細胞凋亡(apoptosis)路徑 8
1.6 老化作用 8
1.6.1 皮膚老化原因 9
1.6.2 皮膚之自由基影響 9
1.6.3 自由基與活性氧的定義及種類 10
1.6.4 DPPH 自由基清除試驗 10
1.6.5 Glutathione 簡介 11
1.6.6 Glutathione peroxidase 11
1.7 發炎反應抑制 11
1.7.1 發炎介質(inflammatory mediators)的作用 11
1.7.2 發炎介質TNF-α 12
1.8 皮膚保濕作用 12
1.8.1 Hadhb 12
1.9 研究動機 13
第二章 實驗材料與方法 14
2.1 實驗材料 14
2.1.1 細胞培養 14
2.1.2 實驗藥品 14
2.2 研究方法 14
2.2.1 細胞評估模式 14
2.2.2 細胞之UV照射 15
2.2.3 黑色素瘤細胞之黑色素含量測定 15
2.2.4 黑色素瘤細胞內之酪胺酸脢活性測定 15
2.2.5 細胞計數 15
2.2.6 MTT assay 細胞存活率測試 15
2.2.7 mouse TNF-α含量測定 16
2.2.8 老鼠試驗評估 16
2.2.9 基因表現分析 16
2.2.10 DPPH自由基清除試驗 18
2.2.11 皮膚之含水量測定 18
第三章 實驗結果與討論 19
論文研究架構 19
3.1 美白效能評估 20
3.1.1 Ectoine對黑色素瘤細胞生長之影響 20
3.1.2 Ectoine對黑色素瘤細胞的黑色素含量影響 23
3.1.3 老鼠黑色素瘤細胞在照射UVA後,ectoine對細胞生長之影響 24
3.1.4 老鼠黑色素瘤細胞在照射UVA後,ectoine對黑色素含量影響 27
3.1.5 Ectoine對細胞內酪胺酸脢活性之影響 28
3.1.6 Ectoine對細胞內細胞凋亡基因表現之影響 30
3.1.7 Ectoine對細胞內酪胺酸脢基因表現之影響 31
3.2發炎反應評估 34
3.2.1 Ectoine對細胞發炎情形之影響 34
3.2.2 Ectoine對細胞內TNF-α基因表現之影響 37
3.3抗氧化作用評估 38
3.3.1 Ectoine之DPPH自由基清除力試驗 38
3.3.2 Ectoine對細胞內氧化基因GPx1表現之影響 39
3.4 保濕效能評估 42
3.4.1 Ectoine對老鼠皮膚含水量之影響 42
3.4.2 Ectoine對細胞內Hydratase基因表現之影響 43
3.5 老鼠皮膚試驗 45
3.5.1 Ectoine對UVA照射之影響 45
3.5.2 Ectoine對UVB照射之影響 47
第四章 結論 49
參考文獻 50


1.Pinnell, S.R., "Regulation of collagen biosynthesis by ascorbic acid:a review. " Yale J Biol Med 58, pp. 553-559, 1985.
2.Galinski,E.A., Pfeiffer,H.P., and Truper,H.G.,             "1,4,5,6-Tetrahydro-2-methyl-4-pyrimidinecarboxylic acid. A novel cyclic amino acid from halophilic phototrophic bacteria of the genus Ectothiorhodospira. " Eur J Biochem 149, pp. 135-139, 1985.
3.Farwick,M., Siewe,R.M., and Kramer,R., "Glycine betaine uptake after hyperosmotic shift in Corynebacterium glutamicum. " J Bacteriol 177, pp. 4690-4695, 1995.
4.Malin,G., and Lapidot,A., "Induction of synthesis of tetrahydropyrimidine derivatives in Streptomyces strains and their effect on Escherichia coli in response to osmotic and heat stress. " J Bacteriol 178, pp. 385-395, 1996.
5.Wohlfarth,A., Severin,J., and Galinski,E.A., "The spectrum of compatible solutes in heterotrophic halophilic eubacteria of the family Halomonadaceae. " J Gen Microbiol 136, pp. 705-702, 1990.
6.Gouesbet,G., Trautwetter,A., Bonnassie,S., Wu,L.F., and Blanco,C.    "Characterization of the Erwinia chrysanthemi osmoprotectant transporter gene ousA. " J Bacteriol 178, pp. 447-455, 1996.
7.Jebbar,M., Talibart,R., Gloux,K., Bernard,T., and Blanco,C.       "Osmoprotection of Escherichia coli by ectoine: uptake and accumulation characteristics. " J Bacteriol 174, pp. 5027-5035, 1992.
8.Talibart,R., Jebbar,M., Gouesbet,G., Himdi-Kabbab,S., Wroblewski,H., Blanco,C., and Bernard,T., "Osmoadaptation in rhizobia: ectoine-induced salt tolerance. " J Bacteriol 176, pp. 5210-5217, 1994.
9.Wondrak,G.T., Jacobson,M.K., and Jacobson,E.L. "Endogenous UVA-photosensitizers: mediators of skin photodamage and novel targets for skin photoprotection. " Photochem Photobiol Sci 5, pp. 215-237, 2006.
10.Toyoda,M., and Bhawan,J. "Ultrastructural evidence for the participation of Langerhans cells in cutaneous photoaging processes: a quantitative comparative study. " J Dermatol Sci 14, pp. 87-100, 1997.
11.Grewe,M. "Chronological ageing and photoageing of dendritic cells." Clin Exp Dermatol 26, pp. 608-612, 2001.
12.Lippert,K., and Galinski,E.A. "Enzyme stabilisation by ectoine-type compatible solutes: protection against heating, freezing and drying. " Appl Microbiol Biotechnol 37, pp. 61-65, 1992.
13.Botta,C., Di,G.C., Sabatier,A.S., and De,M.M. "Genotoxicity of visible light (400-800 nm) and photoprotection assessment of ectoin, L-ergothioneine and mannitol and four sunscreens. " J Photochem Photobiol B 91, pp. 24-34., 2008.
14.Buommino,E., Schiraldi,C., Baroni,A., Paoletti,I., Lamberti,M., De,R.M., and Tufano,M.A. "Ectoine from halophilic microorganisms induces the expression of hsp70 and hsp70B'' in human keratinocytes modulating the proinflammatory response. " Cell Stress Chaperones 10, pp. 197-203, 2005.
15.Buenger,J., and Driller,H. "Ectoin: an effective natural substance to prevent UVA-induced premature photoaging. " Skin Pharmacol Physiol 17, pp. 232-237, 2004.
16.Graf,R., Anzali,S., Buenger,J., Pfluecker,F., and Driller,H. "The multifunctional role of ectoine as a natural cell protectant. " Clin Dermatol 26, pp. 326-333, 2008.
17.Lentzen,G., and Schwarz,T. "Extremolytes: Natural compounds from extremophiles for versatile applications. " Appl Microbiol Biotechnol 72, pp. 623-634, 2006.
18.Wondrak,G.T., Jacobson,M.K., and Jacobson,E.L. "Endogenous UVA-photosensitizers: mediators of skin photodamage and novel targets for skin photoprotection. " Photochem Photobiol Sci 5, pp. 215-237, 2006.
19.Harishchandra RK, Wulff S, Lentzen G, Neuhaus T, Galla HJ. "The effect of compatible solute ectoines on the structural organization of lipid monolayer and bilayer membranes. " Biophys Chem. 150, pp. 37-46., 2010.
20.許元昱,李旺祚,郭文勵譯,組織學,合記圖書出版,1992。
21.徐莉萍,柯哲銘等編譯,ROSS 組織學,合記圖書出版,1991。
22.陳亦瑋, 「石蓮萃取物對洋菇酪胺酸酶活性影響之研究」, 靜宜大學應用化學究所,碩士論文,2002。
23.Thong, H.Y., et al., "The patterns of melanosome distribution in keratinocytes of human skin as one determining factor of skin colour. " Br J Dermatol 149, pp. 498-505,2003.
24.Pawelek J. M. and Korner A. M., "Ther biosynthesis of mammalian melanin. " Amer.Sci. 70, pp. 136-145, 1982.
25.Slominski, A., et al., "Melanin pigmentation in mammalian skin and its hormonal regulation. " Physiol Rev. 84, pp. 1155-1228, 2004.
26.Pinnell, S.R., "Regulation of collagen biosynthesis by ascorbic acid: a review. " Yale J Biol Med 58, pp. 553-559, 1985.
27.Kitt D., "An evaluation of the multiple effects of the antioxidant vitamins.", Trends in food science & technology 8, pp. 198-203,1997.
28.Parvez, S., et al., "Survey and mechanism of skin depigmenting and lightening agents. " Phytother Res. 20, pp. 921-934, 2006.
29.Jenkins R. R., "Free radical chemistry: relationship to exercise sports medicine" 5, pp. 156-170, 1988.
30.Roy C.R., Gies H.P., Lugg D.J., Toomey S., Tomlinson D.W., "The measurement of solar ultraviolet radiation, Mutat. " Res. 422, pp. 7-14, 1998.
31.Diffey B.L., Larko O., "Clinical climatology." Photodermatol 1, pp. 30-37, 1984
32.McCord, JM.; Fridovich, I., "Superoxide dismutase. " J. Biol. Chem. 244, pp. 6049-6055, 1969.
33.Peter K.M. Kim, Richard Weller, Yun Hua, and Timothy R. Billiar., "Ultraviolet irradiation increases FADD protein in apoptotic human keratinocytes." Biochemical and Biophysical Research Communications 302, pp. 290–295, 2003.
34.Dagmar Kulms, Thomas Schwarz., "Independent contribution of three different pathways to ultraviolet-B-induced apoptosis." Biochemical Pharmacology 64, pp. 837-841, 2002.
35.Molecular Cell Biology (Fifth Edition) Copyright © (2004) by W. H. Freeman & Company Chapter 21-23

36.Chung et al., "Modulation of skin collagen metabolism in agedand photoaged human skin in vivo." The Journal of InvestigativeDermatology 117, pp. 1218-1224, 2001.
37.Kochevar, I. E , Moran, M., Lyon, N. Flotte, T., Siebert, E. andGange, W., "Effects of systemic indomethacin, meclizine, and BW755C on chronic ultraviolet B-induced effects in hairless mouse skin." The Society for Investigative Dermatology 2, pp. 186-193,1993.
38.Harman, D., "Free radical theory of aging." Mutation Research 275, pp. 257-266, 1992.
39.Del Maestro, R. F., Thaw, H. H., Bjork, T. J., Planker, M. and Arfors, K., "Free radicals as mediators of tissue injury." Acta Physiologica Scandinavica. 492, pp. 43-57, 1980.
40.Jenkins, G., "Molecular mechanisms of skin ageing." Mech. Ageing 123, pp. 802-810, 2002.
41.洪偉章,李金枝,陳榮秀,化妝品原料及功能,藝軒圖書出版社,臺北,1998。
42.William.J. C., Photoaging. In: H. I. Maibach and P. Elsner(eds.), Cosmeceuticals: Drugs vs. Cosmetics, Marcel Dekker, New York, 2000.
43.Ames, B. N., "Endogeneousb DNA damage as related to cancer and aging. Mutat. " Res. 214, pp. 41-46, 1990.
44.Keith C. B., Lawrence M., "Melanins: hair dyes for the future. " Cosmetics &Toiletries 109, pp. 59-64, 1994.
45.Abeles R. H., Frey P. A., Jenck W. P., Biochemistry, Jones and Bartelett, London, 1991.
46.McCord, JM.; Fridovich, I., "Superoxide dismutase." J. Biol. Chem. 244, pp. 6049-6055, 1969.
47.Aruoma, O. I., "Nutrition and health aspects of free radicals and antioxidants." Food Chem. Toxic. 32, pp. 671-683, 1994.
48.Corrigan, F. M.; Welsh, S. W.; Skinner, E. R.; Horrobin, D. F., "Brain lipid in aging and in Alzheimer’s disease: a review." J. Nutr. Med. 4, pp. 327-349,1994.

49.Halliwell, B.; J. M. C. Gutteridge, C. E. Cross., "Free radical, antioxidants and human disease: where are we now?" J. Lab. Clin. Med. 119, pp. 598-602, 1992.
50.Kinsella, J.; Frankel, E.; German, B. and Kanner, J., "Possible mechanism for the protective role of antioxidants in wine and plant foods." Food Technol. 47, pp. 85-89, 1993.
51.Halliwell, B. and Gutteridge, J.M.C., Free Radicals in Biology and Medicine., 8, pp. 484-487, 1989
52.Simic, M. G., "Mechanisms of inhibition of free-radical processes in mutagenesis and carcinogenesis." Mutat. Res. 202, pp. 377-386,1988
53.Cadenas, E., "Mechanisms of oxygen activation and reactive oxygen species detoxification." Oxidative Stress and Antioxidant Defenses in Biology pp.1-61. Chapman & Hall, New York, USA, 1995.
54.Halliwell, B.; Murcia, M. A.; Chirico, S.; Aruoma, O. I., "Free radicals and antioxidants in food and in vivo: what they do and how they work." Crit. Rev. Food Sci. Nutr. 35, pp. 7-20, 1995.
55.Andreas, M. P. Chemistry of active oxygen species and antioxidants. In “Antioxidant Status, Diet, Nutrition, and Health.” pp. 3-20. CPC Press, Boca Raton London New York Washington, D. C.
56.賴茲漢,賴業超,食品科技辭典,富林出版社,台中。
57.Dinis, T. C. P., Madeira, V. M. C. and Almeida, L. M., "Actionof phenolic derivatives (Acetaminophen,salicylate,and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers." Archives of Biochemistry and Biophysics. 315, pp. 161-169, 1994.
58.King N., "The use of comparative quantitative RT-PCR to investigate the effect of cysteine incubation on GPx1 expression in freshly isolated cardiomyocytes. " 630, pp. 215-32, 2009.
59.Nordlund, J. J.; Abdel-Malek, Z. A., "Mechanisms for post-inflammatory hyperpigmentation and hypopigmentation." In: Advances in Pigment Cell Society, J. Bagnara (ed). 219-236, 1988.
60.Swope, V. B.; Abdel-Malek, Z. A.; Kassen, L.; Nordlund, J. J., "Interkeukins 1 and 6 and tumor necrosis factor are paracrine inhibitors of human melanocyte proliferation and melanogensis." J. Invest. Dermatol. 96, pp. 180-185, 1991.
61.Abdel-Malek, Z.; Swope, V. B.; Collins, C.; Boissy, R.; Zhao, H.; Nordlund, J. J., "Contribution of melanogenic proteins to the heterogeneous pigmentation of human melanocytes. " J. Cell Sci. 106, pp. 1323-1331, 1993.
62.Swppe, V. B.; Sauder, D. N.; McKenzie, R. C.; Sramkoski, R. M.; Krug, K. A.; Babcock, G. F.; Nordlund, J. J.; Abdel-Malek, Z. A., "Synthesis of interkeukin-1? and ? by normal human melanocytes. " J. Invest. Dermatol. 102, pp. 749-753, 1994.
63.Locksley RM, Killeen N, Lenardo MJ., "The TNF and TNF receptor superfamilies: integrating mammalian biology." Cell 104, pp. 487–501, 2001.
64.Heinrich U, Garbe B, Tronnier H., "In vivo Assessment of Ectoin: A Randomized, Vehicle-Controlled Clinical Trial." Skin Pharmacol Physiol. 20, pp. 211-218, 2007.
65.Furuhashi M, Ura N, Murakami H., "Fenofibrate improves insulin sensitivity in connection with intramuscular lipid content, muscle fatty acid-binding protein, and beta-oxidation in skeletal muscle. " J Endocrinol. 174, pp. 321-329, 2002.


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