臺灣博碩士論文加值系統

本研究的目的在評估將萃取自龍鬚菜(Gracilaria lemaneformis )之mycosporine-like amino acids (MAAs) 添加於化妝品，以應用於人體皮膚上之抗紫外光照射皮膚所引起的光老化能力及抗氧化能力。實驗首先分析MAAs之特性，並比較MAAs螯合亞鐵離子、清除DPPH自由基及超氧陰離子之能力，來求出抗光老化化妝品的最適濃度；再以細胞培養方法觀察MAAs對抗紫外線照射引起的光老化效果。之後將其製成含MAAs之乳霜，並探討其安定性及有效性如皮膚彈性、皮膚粗糙度及敏感性之影響。
實驗結果顯示，萃取自龍鬚菜(Gracilaria lemaneformis )之MAAs在75 ℃下受熱2小時，在UVB波長區域內的吸光效果不因受熱而降低，顯示其不會有顯著的衰退或變性現象，因此在製成乳霜時不需擔心熱破壞。
抗氧化方面：濃度在1-2 % MAAs有較佳清除超氧陰離子及DPPH自由基的效果，清除率可達90 %以上；螯合亞鐵離子方面，隨MAAs濃度升高而有較佳螯合能力，2 % MAAs有近50 %的螯合效果。
細胞實驗方面：加入MAAs於培養基濃度達 12.5 μg/ml 以上時，其對正常人類皮膚纖維母細胞(CCD-966SK)合成膠原蛋白有顯著的促進作用；以MAAs培養黑色素腫瘤母細胞(A375)，隨MAAs濃度升高會有較好的抑制黑色素生成效果。
Rt-PCR實驗結果說明，MAAs可存在於正常人類皮膚纖維母細胞內並保持抗紫外線功能，減輕細胞受紫外線刺激後老化機制的進行而達到抗光老化之效果。
含或不含MAAs之乳霜之安定性，經超音波震盪5分鐘、離心30分鐘後不會發生相分離，在50℃中貯存2個月以上皆可維持正常不變質，顯示出該產品具有良好的安定性。
塗抹含MAAs之乳霜後不會有皮膚紅腫現象產生，且六週後對皮膚之彈性值R2及R8皆有增加之情形。在皮膚粗糙度試驗塗抹含MAAs之乳霜六週後，其皮膚鱗片度及皮膚皺紋度指標值也有明顯降低趨勢。

The purpose of the thesis is to explore the efficacy of an anti-photoaging skincare that containing the MAAs extracts of Gracilaria lemaneformis. The UV absorption characteristics of MAAs, the efficacy of anti-aging such as the chelating ferrous ions capacities and the scavenging capacity of DPPH radical and superoxide radical were evaluated to find the optimize concentration of the anti-photoaging skincare. The efficacy of MAAs anti-photoaging in vitro by cells line experimentation was also explored. An anti-photoaging skincare that containing the MAAs extracts was prepared and assay its characteristics including storage stability, safety test and effecacy such as the skin elasticity, skin roughness.
The results obtained are:
1. The absorption spectrum of the MAAs extracts in UVB range didn’t change when heating at 75 ℃ for 2 hours. The result indicated that the photo-protective properties of MAAs will not decrease or denature during heating.
2. The MAAs extracts showed good anti-oxidation capability, the scavenging effects for DPPH radical and superoxide radical are near 90 % at MAAs concentration ranged between 1-2 %. The chelating ferrous ions capacity increased with increasing the concentration of MAAs, is about 50 % at 2 % MAAs.
3. Cells line data shows that the effect of collagen secretion capability of the cultured human normal skin fibroblast increases when the concentration of MAAs is higher than 12.5 μg/ml. The effect of melanin inhibition of the cultured melanocyte increased with increasing the concentration of MAAs.
4. Rt-PCR data shows that the cultured human normal skin fibroblast were granted their sunscreen capability after the cell line were cultured with MAAs and resulted in, some relief against photo-oxidative stress.
5. The cosmetic products containing either with or without MAAs were stable for sonication (135 W, 42KHz, 5 min), for centrifuging (3000 rpm, 30 min), and for more than two months at high temperature (50□C) storage.
6. The safety test resulted in no erythema based on the Daize score test. Skin eleasticity (R2, R8) for those whom have applied the cosmetic product containing MAAs increased over time during six weeks test period. Skin roughness decreased over time during six weeks test period and did show a significant difference for skin scaliness and skin wrinkles index.

中文摘要…..……………………………….………..……………….......1
英文摘要…..……………………………….………..……………….......3
一、前言……………………………………………..………………..…5
二、 文獻整理…..……………………………….………..…………..8
1. 龍鬚菜之來源與利用...……………………….………..…………..8
2. Mycosporine-like amino acids (MAAs)……………………...……..9
2. 1. MAAs之分怖...............................................……..…………….....10
2. 1. 1. 藍藻中的MAAs……………………..………………...…….....10
2. 1. 2. 浮游真核藻類(微藻和大型藻)中的MAAs……………....…...11
2. 1. 3. 細菌和真菌中的MAAs..............................................................11
2. 1. 4. 其他水生生物中的MAAs..........................................................11
3. 皮膚老化之簡介........... .....................................................................11
3. 1. 內在因素……………………………………...…..........................13
3. 2. 外在因素.........................................................................................14
3. 2. 1. 紫外線與皮膚老化之關係.........................................................16
3. 2. 紫外線與色素沉澱之關係.............................................................18
3. 3. 紫外線與皮膚免疫之關係.............................................................19
4. 紫外線與皮膚防曬的關係.................................................................20
4. 1.一般防曬劑之分類...........................................................................20
4. 1. 1. 物理性防曬劑(Physical sunscreens)...........................................21
4. 1. 2. 化學性防曬劑(Chemical sunscreens).........................................21
三、實驗材料…................................................................................23
1. 龍鬚菜之來源.....................................................................................23
2. 細胞株之來源.....................................................................................23
3. 化妝品原料之來源.............................................................................23
四、實驗方法............................................................................................24
1. 龍鬚菜濕藻中MAAs之取得..............................................................24
1. 1. MAAs之甲醇萃取...........................................................................24
1. 2. 高效液相層析技術.........................................................................24
1. 3. MAAs之安定性測試.......................................................................25
1. 3. 1. 抗UV能力測定...........................................................................25
1. 3. 2. 熱穩定性之測定.........................................................................25
2. 細胞培養.............................................................................................25
2. 1. 附著性細胞之培養.........................................................................25
2. 2. 細胞的繼代培養.............................................................................26
2. 3. 細胞解凍與保存.............................................................................26
3. MAAs之抗老化能力評估...................................................................27
3. 1. 試管試驗.........................................................................................27
3. 1. 1. 清除α , α- diphenyl-β-picryhydrazyl (DPPH)自由基能力之測定.................................................................................................27
3. 1. 2. 清除超氧陰離子能力之測定.....................................................28
3. 1. 3. 螯合亞鐵離子能力之測定.........................................................29
3. 2. 細胞實驗.........................................................................................30
3. 2. 1. MAAs對於纖維母細胞存活率之影響.......................................30
3. 2. 1. 1. MTT assay................................................................................31
3. 2. 2. MAAs對促進膠原蛋白合成之作用...........................................31
3. 2. 3. MAAs對抑制黑色素細胞生成黑色素的影響...........................32
3. 3. MAAs抗光老化之基因表現測定...................................................32
3. 3. 1. 細胞前處理.................................................................................33
3. 3. 2. Total RNA 之抽取......................................................................33
3. 3. 2. 以One step SYBR Green real-time RT-PCR檢測mRNA含量....34
4. MAAs活力滋潤霜之製備...................................................................35
5. MAAs活力滋潤霜之品質測試...........................................................36
5. 1. 安定性測試.....................................................................................36
5. 1. 1. 儲存溫度測試.............................................................................36
5. 1. 2. 色度測試 L*a*b*.....................................................................36
5. 1. 3. 離心機分離試驗.........................................................................36
5. 1. 4. 超音波震盪試驗.........................................................................36
5. 2. 安全性測試.....................................................................................37
5. 2. 1. 貼布試測法(patch test)...............................................................37
5. 2. 2. 皮膚pH值之測定......................................................................37
6. MAAs活力滋潤霜對皮膚特性之影響...............................................37
6. 1. 塗抹前後六星期以吸允法測定手臂皮膚的彈性.........................37
6. 2. 對皮膚粗糙度之in vivo測定..........................................................37
7. 統計分析.............................................................................................38
五、結果與討論.......................................................................................39
1. 龍鬚菜濕藻中MAAs之取得..............................................................39
1. 1. MAAs高效液相層析技術結果.......................................................39
1. 2. MAAs之安定性測試.......................................................................39
1. 2. 1. 抗UV能力測定...........................................................................39
1. 2. 2. 熱穩定性之測定.........................................................................41
2. MAAs之抗老化能力評估...................................................................41
2. 1. 試管試驗.........................................................................................43
2. 1. 1. 清除α , α- diphenyl-β-picryhydrazyl (DPPH)自由基能力之測定................................................................................................43
2. 1. 2. 清除超氧陰離子能力之測定.....................................................43
2. 1. 3. 螯合亞鐵離子能力之測定.........................................................44
2. 2. 細胞實驗.........................................................................................45
2. 2. 1. MAAs對於纖維母細胞存活率之影響.......................................45
2. 2. 2. MAAs對促進正常人類皮膚纖維母細胞膠原蛋白合成之作用
......................................................................................................46
2. 2. 3. MAAs對抑制黑色素細胞生成黑色素的影響...........................46
2. 2. 4. MAAs 抗光老化之基因表現測定.............................................47
2. 2. 4. 1. MMP-1及MMP-2 mRNA之表現............................................49
2. 2. 4. 2. TIMP-1 mRNA之表現.............................................................49
2. 2. 4. 3. p53 mRNA之表現....................................................................50
3. 含MAAs之活力滋潤霜品質特性與人體試驗分析..........................51
3. 1. 品質特性.........................................................................................5 1
3. 1. 1. 安定性.........................................................................................51
3. 1. 2. 色度測試 L*a*b*.....................................................................51
3. 2. 對皮膚之安全性.............................................................................51
3. 2. 1. 皮膚敏感性.................................................................................51
3. 3. 對皮膚之有效性.............................................................................52
3. 3. 1. 皮膚彈性.....................................................................................52
3. 3. 2. 皮膚鱗片度.................................................................................53
3. 3. 3. 皮膚皺紋度.................................................................................53
六、結論..................................................................................................55
七、參考文獻..........................................................................................56
表………………………………………….……………........……….....67
圖……………………………………………………….......……….......74
附錄……………….......……….........................................................91

方銘志，2002，山藥多醣體之免疫調節機能探討，國立台灣海洋大學食品科學系碩士論文。
化妝品衛生管理條例暨有關法規，2004，行政院衛生署。
光井武夫（陳韋達、鄭慧文譯），1992，新化妝品學，南山堂株式會社出版，合記圖書出版社發行，13-46。
自然保育季刊 2001.3 第三十三期 54-55
林東明，1993，用於食品及化妝品的紅微細藻天然色素，今日經濟，1 月號：79-85。
倪志文，2000，剪力與活性氧族群對MMP-1與TIMP-1表現之調控，國立台灣大學化學工程學研究所碩士論文。
馬盈瑜，2003，龍鬚菜多醣及其水解產物生理活性之探討，國立屏東科技大學食品科學系碩士學位論文。
黃霨禎，2005，龍鬚菜寡醣水解物之製備及其抗氧化與抗致突變性之探討與對雙叉桿菌生長影響，國立台灣海洋大學食品科學系碩士論文。
張成武和殷志敏，1995，藻膽蛋白的特色與利用，中國海洋藥物雜志，3：52-53。
張思維，2004，開發隔離紫外光的產品及其安定性之研究，朝陽科技大學應用化學系碩士論文。
陳宜嫻，2001，皮膚黑色素抑制劑與保濕劑效能之研究，靜宜大學應用化學研究所碩士論文。
陳宣旭，2004，氧化逆境下血管內皮細胞中基質金屬蛋白酵素-2(MMP-2)之表現：活化轉錄因子3(ATF3)所扮演之角色，國防醫學院生命科學研究所博士論文。
廖姿婷，2002，龍鬚菜抗氧化性質評估及其飲料產品研製，國立中興大學食品科學系碩士學位論文。
廖保鑫，1994，口腔黏膜下纖維化細胞、口腔癌細胞株p53的表達與細胞凋亡的探討，中山醫學大學醫學研究所博士論文。
劉正文，鐘平，韓博平，銅綠微囊藻中的紫外光保護物質類菌孢素胺基酸(MAAs)與水華形成機制探討，湖泊科學，15：359-363。
劉錦芬，2001，臺灣養殖龍鬚菜萃取物之免疫生理活性探討，國立臺灣海洋大學食品科學系碩士學位論文。
蔡心涵，1995，螺旋藻藻藍蛋白對癌激發療法增敏作用的實驗研究，中國海洋藥物雜志，1：15-18。
蔡昆男，2001，藻膽蛋白抗腸病毒71型之能力研究，長庚大學基礎醫學研究所碩士論文。
蔡季真，2005，油脂與乳化劑型對防曬產品的SPF值之影響，靜宜大學應用化學研究所碩士論文。
鐘逸甫，2003，吃的防曬油-海洋動物抗紫外線的法寶，自然保育季刊，33: 54-55。
Allen, J. B. 2001. Molecular mechanisms of aging in connective tissues. Mechanisms of Ageing and Development. 122: 735-755.
Ashcroft, G. S. 1997. Human ageing impairs injury-induced in vivo expression of tissue inhibitor of matrix metalloproteinases (TIMP)-1 and -2 proteins and mRNA. American Journal of Pathology. 183: 169-76.
Bandaranayake, W. M. 1998. Mycosporines: Are they nature's sunscreens? Natural Product Reports.15:159-172.
Bian, J. and Y. Sun. 1997. Transcriptional activation by p53 of the human type IV collagenase (gelatinase A or matrix metalloproteinase 2) promoter. Molecular and Cellular Biology. 17: 6330-8.
Blease, K., Schuh, J. M., Jakubzick, C., Lukaces, N. W., Kunkel, S. T., Joshi, B. H., Puri, R. K., Kaplan, M. H., and Hogaboam, C.M. 2002. State-deficient mice develop airway hyperresponsiveness and peribronchial fibrosis during chronic fungal asthma. American Journal of Pathology. 160: 481-490.
Bonina, F., Saija, A., Tomaino, A., Cascio, R. L., Rapisarda, P. And, Dederen, J. C. 1998. In vitro antioxidant activity and in vivo photoprotective effect of a red orange extract. International Journal of Cosmetic Science. 20: 331-342.
Brenneisen, P., Briviba, K., Wlaschek, M., Wenk, J., and, Scharffetter-Kochanek K. 1997. Hydrogen peroxide increases the steady-state mRNA levels of 80 collagenase/MMP-1 in human dermal fibroblasts. Free Radical Biology and Medicine. 22: 515-524.
Brenneisen, P., Wenk, J., Klotz, L. O., Wlaschek, M., Briviba, K., Krieg, T., Sies, H., and Scharffetter-Kochanek, K. 1998. Central role of Ferrous/Ferric iron in the ultraviolet B irradiation-mediated signaling pathway leading to increased interstitial collagenase (matrix-degrading metalloprotease (MMP)-1) and stromelysin-1 (MMP-3) mRNA levels in cultured human dermal fibroblasts. The Journal of Biological Chemistry. 273: 5279-5287.
Callone, A. I., Carignan, M., Montoya, N. G., and Carreto, J. I. 2006. Biotransformation of mycosporine like amino acids (MAAs) in the toxic dinoflagellate Alexandrium tamarense. Journal of Photochemistry and Photobiology B: Biology. 84: 204-212.
Castenholz, R.W., and Garcia-Pichel, F. 2000. Cyanobacterial responses to UV-radiation. In The Ecology of Cyanobacteria: Their Diversity in Time and Space. Dordrecht:Kluwer Academic Publications. pp. 591-611.
Conde, F. R., Churio, M. S., and Previtali, C. M. 2000. The photoprotector mechanism of mycosporine-like amino acids. Excited-state properties and photostability of porphyra-334 in aqueous solution. Journal of Photochemistry and Photobiology. 56: 139-144.
Dunlap, W. C., Williams D. M., Chalker B. E., and Banaszak A. T. 1989. Biochemical photoadaptations in vision: UV-absorbing pigments in fish eye tissues. Comparative Biochemistry and Physiology. 93: 601–607.
Dinins, T. C. P., Maderia, V. M. C., and Almeida, L. M. 1994. Action of phenolic derivatives (acetaminophen, salicylate and 5- aminosalicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers. The Journal of Investigative Dermatology. 100: 692-698.
Dunlap, W. C., and Shick, J. M. 1989. Ultraviolet radiation-absorbing mycosporine-like amino acids in coral reef organisms: a biochemical and environmental perspective. Journal of Phycology. 34: 418-430.
Dunlap, W. C. and Yamamoto, Y. 1995. Small-molecule antioxidants in marine organisms: antioxidant activity of mycosporine-glycine. Comparative Biochemistry and Physiology. 1: 105-114.
Dunlap, W. C., Masaki, K., Yamamoto, Y., Larsen, R. M., and Karube, I. 1998. A novel antioxidant derived from seaweed. In New Developments in Marine Biotechnology. New York:Plenum. pp. 33-35.
Dunlap, W. C., Shick, J. M., and Yamamoto, Y. 2000. UV protection in marine organisms. London: International Organization of Motor Vehicle Manufacturers Internationalias. pp. 200-214.
Ellerbroek, S. M., and Stack, M. S. 1999. Membrane associated matrix metalloproteinases in metastasis. Bioessays. 21: 940-949.
Eric, F. B., and Jouni, U. 1996. The effect of photodamage on dermal extracellular matrix. Clinical Dermatology. 14: 143-151.
Fernandez-Patron, C., Zouki, C., Whittal, R. M., Chan John S.D., Davidge S. T., and Filep J. G. 2002. Methods for analysis of matrix metalloproteinase regulation of neutrophil-endothelial cell adhesion. Biological Procedures online. 4: 38-48.
Figueroa, F. L., Escassi, L., Perez-Rodrýguez, E., and Korbee, N. 2003. Effects of short-term irradiation on photoinhibition and accumulation of mycosporine-like amino acids in sun and shade species of the red algal genus Porphyra. Journal of Photochemistry and Photobiology. 69: 21–30.
Garcia-Pichel, F., Wingard, C. E., and Castenholz, R.W. 1993. Evidence regarding the UV sunscreen role of a mycosporine-like compound in the cyanobacterium Gloeocapsa sp. Applied and Environmental Microbiology. 59: 170-176.
Goihman, Y. M. 1996. Skin aging and photoaging: an outlook. Clinical Dermatology. 14: 153-160.
Gomez, D. E., Alonso, D. F., Yoshiji, H., and Thorgeirsson, U. P. 1997. Tissue inhibitors of metalloproteinases: structure, regulation and biological functions. European Journal of Cell Biology. 74: 111-122.
Hahhiew, I. M., Eller, M. S., and Gilchrest, B. A. 2000. Skin aging and photoaging: the role of DNA damage and repair. American Journal of Contact Dermatitis. 1: 19-25.
Hazane, F., Valenti, K., Sauvaigo, S., Peinnequin, A., Mouret, C., Favier, A., and Beani, J. 2005. Ageing effects on the expression of cell defence genes after UVA irradiation in human male cutaneous fibroblasts using cDNA arrays. Journal of Photochemistry and Photobiology. B: Biology. 79: 171-190.
Hoyer, K., Karsten, U., Sawall, T., and Wiencke, C. 2001. Photoprotective substances in Antarctic macroalgae and their variation with respect to depth distribution, different tissues and developmental stages. Marine Ecology Progress series. 211: 117-129.
Jeffrey, S. W., MacTavish, H. S., Dunlap, W. C., Vesk, M., and Groenewoud, K. 1999. Occurrence of UVA- and UVB-absorbing compounds in 152 species (206 strains) of marine microalgae. Marine Ecology Progress Series. 189: 35–51.
Jiao, H., Soejima, Y., Ohe, Y., and Saijo, N. A. 1992. A new MTT assay for examining the cytotoxicity of activated macrophages towards the nonadherent P388 leukemia cell line. Journal of Immunological Methods. 153: 265-266.
John, G.S. 1992. Molecular biology of free radical scanvenging system. pp. 47-75.
Kanoh, H., Kitamura, T., and Kobayashi, Y. 1992. A sulfate proteoglycan from the red alga Gracilaria verrucosa is a hemagglutinin. Comparative Biochemistry and Physiology. 102: 445-449.
Karsten, U., Franklin, L.A., Lüning, K., and Wiencke, C. 1998. Natural ultraviolet radiation and photosynthetically active radiation induce formation of mycosporine-like amino acids in the marine macroalga Chondrus crispus (rhodophyta). Planta. 205: 257-262.
Karsten, U., and West J.A. 2000. Living in the intertidal zone — seasonal effects on heterosides and sun-screen compounds in the red alga Bangia atropurpurea (Bangiales). Journal of Experimental Marine Biology and Ecology. 254: 221-234.
Korbee, N., Figueroa, F. L., and Aguilera, J. 2005. Effect of light quality on the accumulation of photosynthetic pigments, proteins and mycosporine-like amino acids in the red alga Porphyra leucosticta (Bangiales, Rhodophyta). Journal of Photochemistry and Photobiology. 80: 71-78.
Karsten, U., Sawall, T., Hanelt, D., Bischof, K., Figuero, F., Flores-Moya, L., and Wiencke, A. 1998a. An inventory of UV-absorbing mycosporinelike amino acids in macroalgae from polar to warmtemperate regions. Marine Botany. 41: 443-453.
Karsten, U., Sawall, T., and Wiencke, C. 1998b. A survey of the distribution of UV-absorbing substances in tropical macroalgae. Phycologia Research. 46: 271-279.
Klampfl, C. W., and Leitner, T. 2003. Quantitative determination of UV filters in sunscreen lotions using microemulsion electrokinetic chromatography. Journal of Separation Science. 26: 1259-1262.
Klisch, M., and Häder, D. 2000. Mycosporine-like amino acids in the marine dinoflagellate Gyrodinium dorsum: induction by ultraviolet irradiation. Journal of Photochemistry and Photobiology. B:Biology. 55: 178-182.
Lahmann, C., Young A. R., Wittern, K., and Bergemann J. 2001. Induction of mRNA for matrix metalloproteinase 1 and tissue inhibitor of metalloproteinases 1 in human skin in vivo by solar simulated radiation. Photochemistry and Photobiology. 73: 657-663.
Lenton, K. J., and Greenstock, C.L. 1999. Ability of human plasma to protect against ionising radiation is inversely correlated with age. Mechanisms of Ageing and Development. 107: 15-20。
Li, Y. Y., Feng, Y., McTiernan, C. F., Pei, W., Moravec, C. S., Wang, P., Rosenblum, W., Kormos, R. L., and Feldman, A. M. 2001. Downregulation of matrix metalloproteinases and reduction in collagen damage in the failing human heart after support with left ventricular assist devices. Circulation. 104: 1147-1152.
Liu, F., and Ng, T. B. 1999. Antioxidative and free radical scavenging activities of selected medicinal herbs. Life Sciences. 66: 725-735.
Mason, D. S., Schafer, F., Shick, J. M., and Dunlap, W. C. 1998. Ultraviolet radiationabsorbing mycosporine-like amino acids (MAAs) are acquired from their diet by Medaka fish (Oryzias latipes) but not by SKH hairless mice. Comparative Biochemistry and Physiology. 120: 587-598.
McKay, B. C., and Ljungman, M. 1999. Role for p53 in the recovery of transcription and protection against apoptosis induced by ultraviolet light. Neoplasia. 1: 276-284.
Nahm, W. K., Zhou, L., and Falanga, V. 2002. Sustained ability for fibroblast outgrowth from stored neonatal foreskin: a model for studying mechanisms of fibroblast outgrowth. Journal of Dermatological Science. 28: 152-158.
Okada, Y. 1990. Matrix metalloproteinase 2 from human rheumatoid synovial fibroblasts. Purification and activation of the precursor and enzymic properties. European Journal of Biochemistry. 194: 721-30.
Oren, A. 1997. Mycosporine-like amino acids as osmotic solutes in a community of halophilic cyanobacteria. Geomicrobiology Journal. 14: 231-240.
O'Shea, M., Willenbrock, F., Williamson, R. A., Cockett, M. I., Freedman, R. B., Reynolds, J. J., Docherty, A. J., and Murphy, G. 1992. Site-directed mutations that alter the inhibitory activity of the tissue inhibitor of metalloproteinases-1: importance of the N-terminal region between cysteine 3 and cysteine 13. Biochemistry. 31: 10146-10152.
Phillips, B. J. 1996. Development of cell culture techniques for assessment of the toxicity of plant products. Toxicology in vitro. 10: 69-76.
Rabe, J. H., Mamelak, A. J., McElgunn, P. J. S., Morison, W. L., and Sauder, D. N. 2006. Photoaging: Mechanisms and repair. Journal of the American Academy of Dermatology. 55: 1-19.
Rajagopalan, S., Meng, X. P., Ramasamy, S., Harrison, D. G., and Galis, Z. S. 1996. Reactive oxygen species produced by macrophage-derived foam cells regulate the activity of vascular matrix metalloproteinases in vitro. Implications for atherosclerotic plaque stability. The Journal of Clinical Investigation. 98: 2572-2579.
Rimbau, V., Camins, A., Romay, C., Gonzalez, R., and Pallas, M. 1999. Protective effects of C-phycocyanin against kainicacid-induced neuronal damage in rat hippocampus. Neuroscience Letters. 276: 75-78。
Robak, J., and Gryglewski, R. J. 1988. Flavonoids are scavengers of superoxide anions. Biochemical Pharmacology. 17: 837-841.
Rozema, J., van de Staaij, J., Bjorn, L. O., and Caldwell, M. 1997. UV-B as an environmental factor in plant life: Stress and regulation. Trends in Ecology and Evolution.12: 22-28.
Sen, C. K., and Packer, L. 1996. Antioxidant and redox regulation of gene transcription. The FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology. 10: 709-720.
Shick, J. M., Dunlap, W. C., Chalker, B. E., Banaszak, A. T., and Rosenzweig, T. K. 1992. Survey of ultraviolet radiation absorbing mycosporinelike amino acids in organs of coral reef holothuroids. Marine Ecology Progress series. 90: 139-148.
Sinha, R.P., Klisch, M., Gröniger, A., and Häder, D. P. 1998. Ultraviolet-absorbing/screening substances in cyanobacteria, phytoplankton and macroalgae. Journal of Photochemistry and Photobiology. 47: 83-94.
Sinha, R. P., Klisch, M., Gröniger, A., and Häder, D. P. 2000. Mycosporine-like amino acids in the marine red alga Gracilaria cornea－effects of UV and heat. Environmental and Experimental Botany. 43: 33-43.
Sommaruga, R., and Garcia-Pichel, F. 1999. UV-absorbing compounds, in planktonic and benthic organisms from a high-mountain lake. Archiv für Hydrobiologie. 144: 225-269.
Südel, K. M., Venzke, K., Knusmann-Hartig, E., Moll, I., Stäb, F., Wenck, H., Wittern, K., Gercken, G., and Gallinat, S. 2003. Tight control of matrix metalloproteinase-1 activity in human skin. Photochemistry and Photobiology. 78: 355-360.
Tartarotti, B., Laurion, I., and Sommaruga, R. 2001. Large variability in the concentration ot mycosporine-like amino acids among zooplankton from lakes located across an altitude gradient. Limnology and Oceanography. 46: 1546-1552.
Tsujino. 1961. I. Studies on the compounds specific for each group of marine algae.Ⅱ.Extraction and isolation of characteristic ultraviolet absorbing material in Rhodophyta. Bulletin of the Faculty Fisheries, Hokkaido University. 12: 59-65.
Volkmann, M., and Gorbushina, A. A. 2006. Abroadlyapplicablemethod forextractionand characterization of mycosporines and mycosporine-like amino acids of terrestrial, marineand freshwater origin. FEMS Microbiology Letters. 255: 286-295.
Wang, S., and Lee, W. 2003. Determination of benzophenones in a cosmetic matrix by supercritical fluid extraction and capillary electrophoresis. Jouranl of Chromatography. 987: 269-275.
Wenk, J., Brenneisen, P., Wlaschek, M., Poswig, A., Briviba, K., Oberley, T. D., Scharffetter-Kochanek, K. 1999. Stable overexpression of manganese superoxide dismutase in mitochondria identifies hydrogen peroxide as a major oxidant in the AP-1-mediated induction of matrix-degrading metalloprotease-1. The Journal of Biological Chemistry. 274: 25869-25876.
Xiong, F., Kopecky, J., and Nedbal, L. 1999. The occurrence of UV-B absorbing mycosporine-like amino acids in freshwater and terrestrial microalgae. Aquatic Botany. 63: 37-49.
Xiong, F., Komenda, J., Kopecky, J., and Nedbal, L. 1997. Strategies of ultraviolet-B protection in microscopic algae. Physiologia Plantarum. 100: 378-388.
Yamamoto, T., and Nishioka, K. 2001. Animal model of sclerotic skin. Ⅳ: induction of dermal sclerosis by bleomycin is T cell independent. Journal of Investigative Dermatology. 117:999-1001.
Yazan, Y., Seiller, M., and Arslan, K. 1997. Formulation and evaluation of a multiple emulsion containing glycolic acid. Drug and Cosmetic Industry. 30: 101-103.
Yoshiki, M. 1995. Photoaging from an oxidative standpoint. Journal of Dermatological Science. 9: 79-86.
Yoshizawa, Y., Tsunehiro, J., Nomura, K., Itoh, M., Fukui, F., Ametani, A., and Kaminogawa, S. l996. In vivo macrophage-stimulation activity of the enzyme-degraded water-soluble polysaccharide fraction from a marine alga (Gracilaria verrucosa). Bioscience Biotechnology Biochemistry. 60: 1667-1671.