臺灣博碩士論文加值系統

隨著時代的進步，人體內在及外在氧化壓力 (Oxidative stress) 都逐漸增加，導致皮膚老化速度加快，因而近年來具抗氧化功效宣稱之植物萃取已逐漸成為皮膚保養品的主流之一。兔尾草 (Uraria crinita) 根部已被廣泛應用於身體保健，但較少作為化妝品中美容護膚用途，莖及葉子的應用更屬罕見，因此本研究目的是將兔尾草根、莖及葉子萃取物分別進行成分鑑定、抗氧化及美白功效評估。兔尾草根、莖及葉子使用乙酸乙酯 (Ethyl Acetate) 萃取，成分鑑定結果顯示三種萃取物皆含有牡荊素 (Vitexin)，而根和莖都測得金雀異黃酮 (Genistein)。三種萃取物的總酚含量分別為20.10 ± 0.87、34.83 ± 0.31、30.80 ± 0.24 mg Gallic acid equivalent (GAE) /g extract，類黃酮含量分別為1.21 ± 0.09、8.16 ± 0.23、19.83 ± 3.05 mg Quercetin equivalent (QE) /g extract，清除DPPH自由基能力IC50分別為3.14 ± 0.05、1.28 ± 0.00、1.25 ± 0.03 mg/mL，還原力分別為24.44 ± 0.33 、29.86 ± 1.20、50.32 ± 2.88 mg Vitamin C equivalent (VCE) /g extract，清除超氧陰離子能力IC50分別為0.10 ± 0.01、0.08 ± 0.01、0.31 ± 0.02 mg/mL，根、莖及葉子萃取物在 ≤ 0.2 mg/mL下對人類角質細胞HaCaT與人類真皮層纖維母細胞Hs68都無細胞毒性，萃取物皆對抑制酪胺酸酶活性及螯合亞鐵離子顯示無效。綜合上述，三種萃取物皆顯示出很好的抗氧化功效，本研究也首度發現根及莖在清除超氧陰離子的功效都明顯較Vitamin C (Vit. C) 來的好，除葉子外，未來根和莖亦可應用於皮膚保養品。

With the progress of the times, the internal and external oxidative stress of body is gradually increasing, resulting in accelerating skin aging. Therefore, in recent years, plant extracts claimed with antioxidant effects have gradually become one mainstream of skin care products. The root of Uraria crinita which has been widely used in health care is less used in cosmetics for skin care, and is even rarer in application of its stem and leaves. Therefore, the purpose of this study was to analyze the chemical composition and to evaluate the antioxidant and whitening efficacy of the root, stem, and leaf extracts of Uraria crinita. The roots, stems, and leaves of Uraria crinita were extracted with ethyl acetate. The compositional identification results showed that all three extracts contained Vitexin, while Genistein was only detected in the roots and stems. The total phenolic contents of the root, stem, and leaf extracts extracts were 20.10 ± 0.87, 34.83 ± 0.31, 30.80 ± 0.24 mg Gallic acid equivalent (GAE) /g extract respectively , and the flavonoid contents were 1.21 ± 0.09, 8.16 ± 0.23, 19.83 ± 3.05 mg Quercetin equivalent (QE) /g extract respectively, the IC50 of scavenging DPPH free radicals were 3.14 ± 0.05, 1.28 ± 0.00, 1.25 ± 0.03 mg/mL respectively, and the reducing powers were 24.44 ± 0.33, 29.86 ± 1.20, 50.32 ± 2.88 mg Vitamin C equivalent (VCE) /g extract respectively, the superoxide anion scavenging ability IC50 was 0.10 ± 0.01, 0.08 ± 0.01, 0.31 ± 0.02 mg/mL respectively. All three extracts had no cytotoxicity on human keratinocyte (HaCaT) and human fbroblast (Hs68) at ≤ 0.2 mg/mL, and the extracts showed no effect on inhibiting tyrosinase activity and chelating ferrous ions. In general, the three extracts all exbited good antioxidant effects. This study also found for the first time that both roots and stems were significantly more effective in scavenging superoxide anions than Vitamin C (Vit. C). In addition to leaves, roots and stems can also be used in skin care products in the future.

致謝 I
中文摘要 II
Abstract III
目錄 V
表目錄 VIII
圖目錄 IX
第一章 緒論 1
1-1 前言 1
1-2 皮膚老化的理論與抗氧化作用 1
1-2-1 皮膚構造 1
1-2-1-1 表皮層 (Epidermis) 3
1-2-1-2 真皮層 (Dermis) 4
1-2-1-3 皮下組織 (Subcutaneous Layer) 4
1-2-2 皮膚老化的理論 5
1-2-3 抗氧化劑的作用 5
1-2-4 類黃酮 (Flavonoids) 介紹 6
1-2-4-1 黃酮 (Flavones) 8
1-2-4-2 黃酮醇 (Flavonols) 8
1-2-4-3 黃烷酮 (Flavanones) 9
1-2-4-4 異黃酮類 (Isoflavonoids) 9
1-2-4-5 花青素 (Anthocyanins) 9
1-2-4-6 查爾酮 (Chalcones) 10
1-3 兔尾草的簡介 10
1-4 研究動機及目的 13
第二章 材料與方法 14
2-1 研究架構 14
2-2 實驗材料 15
2-3 實驗儀器 16
2-4 實驗方法 17
2-4-1 兔尾草萃取物製備 17
2-4-2 成分分析 17
2-4-3 總酚含量測定 18
2-4-4 類黃酮含量測定 19
2-4-5 清除DPPH自由基能力測定 20
2-4-6 螯合亞鐵離子能力測定 21
2-4-7 還原力測定 22
2-4-8 清除超氧陰離子能力測定 23
2-4-9 抑制蘑菇酪胺酸酶活性試驗 24
2-4-10 抑制人類酪胺酸酶活性試驗 25
2-4-11 HaCaT細胞存活率試驗 26
2-4-12 Hs68細胞存活率試驗 27
2-4-13 統計分析 27
第三章 結果與討論 28
3-1 兔尾草萃取物之產率 28
3-2 兔尾草萃取物之成分分析 29
3-3 兔尾草萃取物之總酚含量及類黃酮含量測定 31
3-4 兔尾草萃取物之抗氧化活性測定 32
3-4-1 清除DPPH能力測定 32
3-4-2 螯合亞鐵離子能力測定 34
3-4-3 還原力測定 35
3-4-4 清除超氧陰離子能力測定 37
3-5 兔尾草萃取物之美白活性測定 39
3-5-1 兔尾草萃取物之抑制蘑菇酪胺酸酶活性 39
3-5-2 兔尾草萃取物之抑制人類酪胺酸酶活性 40
3-6 兔尾草萃取物之細胞存活率測定 41
第四章 結論 43
參考文獻 44

Mintel. (2021). Function flourishes in bpc market: 74% of urban chinese female consumers pay attention to specific functional ingredients. https://www.mintel.com/press-centre/beauty-and-personal-care/function-flourishes-in-bpc-market-74-of-urban-chinese-female-consumers-pay-attention-to-specific-functional-ingredients
Fact, MR. (2021). Plant-based Skincare Products Market. https://www.factmr.com/report/plant-based-skincare-products-market
Amaral, A. B., Silva, M. V. d., Lannes, S. C. d. S. (2018). Lipid oxidation in meat: mechanisms and protective factors–a review. Food Science and Technology, 38, 1-15.
Aoki, T., Akashi, T., Ayabe, S.-I. (2000). Flavonoids of leguminous plants: structure, biological activity, and biosynthesis. Journal of Plant Research, 113(4), 475.
Baroni, A., Buommino, E., De Gregorio, V., Ruocco, E., Ruocco, V., Wolf, R. (2012). Structure and function of the epidermis related to barrier properties. Clinics in Dermatology, 30(3), 257-262.
Batovska, D. I., Todorova, I. T. (2010). Trends in utilization of the pharmacological potential of chalcones. Current Clinical Pharmacology, 5(1), 1-29.
Bissett, D., Miyamoto, K., Sun, P., Li, J., Berge, C. (2004). Topical niacinamide reduces yellowing, wrinkling, red blotchiness, and hyperpigmented spots in aging facial skin 1. International Journal of Cosmetic Science, 26(5), 231-238.
Bondet, V., Brand-Williams, W., Berset, C. (1997). Kinetics and mechanisms of antioxidant activity using the DPPH. free radical method. LWT-Food Science and Technology, 30(6), 609-615.
Campos Chiste, R., Freitas, M., Zerlotti Mercadante, A., Fernandes, E. (2015). Superoxide anion radical: generation and detection in cellular and non-cellular systems. Current Medicinal Chemistry, 22(37), 4234-4256.
Chao, J., Dai, Y., Cheng, H.-Y., Lam, W., Cheng, Y.-C., Li, K., Peng, W.-H., Pao, L.-H., Hsieh, M.-T., Qin, X.-M., Lee, M.-S. (2017). Improving the concentrations of the active components in the herbal tea ingredient, Uraria crinita: The effect of post-harvest oven-drying processing. Scientific Reports, 7(1), 1-16.
Chaudhari, S. Y., Ruknuddin, G., Prajapati, P. (2016). Ethno medicinal values of Citrus genus: A review. Medical Journal of Dr. DY Patil University, 9(5), 560.
Cheeseman, K. H., Slater, T. F. (1993). An introduction to free radical biochemistry. British medical Bulletin, 49(3), 481-493.
Chiu, N.-Y., Chang, K.-H. (1992). The illustrated medicinal plants of Taiwan. Southern Materials Center, 3.
Choe, S.-Y., Hong, J.-H., Gu, Y.-R., Kim, I.-D., Dhungana, S., Moon, K.-D. (2019). Hot water extract of Glehnia littoralis leaf showed skin-whitening and anti-wrinkle properties. South African Journal of Botany, 127, 104-109.
Cichorek, M., Wachulska, M., Stasiewicz, A., Tymińska, A. (2013). Skin melanocytes: biology and development. Advances in Dermatology and Allergology/Postȩpy Dermatologii I Alergologii, 30(1), 30.
Dai, T. D., Thien, D. D., Tam, N. T. (2021). Flavone glycosides from Uraria crinita. Vietnam Journal of Science, Technology and Engineering, 63(2), 3-6.
Freedberg, I. (1993). Epidermal differentiation and keratinazation. Dermatology in General Medicine, 198-209.
Haftek, M., Mac‐Mary, S., Bitoux, M. A. L., Creidi, P., Seité, S., Rougier, A., Humbert, P. (2008). Clinical, biometric and structural evaluation of the long‐term effects of a topical treatment with ascorbic acid and madecassoside in photoaged human skin. Experimental Dermatology, 17(11), 946-952.
Harman, D. (2002). Aging: a theory based on free radical and radiation chemistry. Science of Aging Knowledge Environment, 2002(37), 14.
Heim, K. E., Tagliaferro, A. R., Bobilya, D. J. (2002). Flavonoid antioxidants: chemistry, metabolism and structure-activity relationships. The Journal of Nutritional Biochemistry, 13(10), 572-584.
Hossain, H., Shahid-Ud-Daula, A., Jahan, I. A., Nimmi, I., Hasan, K., Haq, M. M. (2012). Evaluation of antinociceptive and antioxidant potential from the leaves of Spilanthes paniculata growing in Bangladesh. Int J Pharm Phytopharmacol Res, 1(4), 178-186.
Irrera, N., Pizzino, G., D’Anna, R., Vaccaro, M., Arcoraci, V., Squadrito, F., Altavilla, D., Bitto, A. (2017). Dietary management of skin health: The role of genistein. Nutrients, 9(6), 622.
Ivey, K. L., Lewis, J. R., Prince, R. L., Hodgson, J. M. (2013). Tea and non-tea flavonol intakes in relation to atherosclerotic vascular disease mortality in older women. British Journal of Nutrition, 110(9), 1648-1655.
Iwashina, T. (2013). Flavonoid properties of five families newly incorporated into the order Caryophyllales. Bull Natl Mus Nat Sci, 39, 25-51.
Kamiloglu, S., Sari, G., Ozdal, T., Capanoglu, E. (2020). Guidelines for cell viability assays. Food Frontiers, 1(3), 332-349.
Khavkin, J., Ellis, D. A. (2011). Aging skin: histology, physiology, and pathology. Facial Plastic Surgery Clinics, 19(2), 229-234.
Klaassen, C. D. (2013). Casarett and Doull's toxicology: The Basic Science of Poisons (Vol. 1236): McGraw-Hill New York.
Kozlowska, A., Szostak-Wegierek, D. (2014). Flavonoids-food sources and health benefits. Roczniki Państwowego Zakładu Higieny, 65(2).
Kruk, I., Aboul‐Enein, H. Y., Michalska, T., Lichszteld, K., Kładna, A. (2005). Scavenging of reactive oxygen species by the plant phenols genistein and oleuropein. Luminescence: The Journal of Biological and Chemical Luminescence, 20(2), 81-89.
Lee, S.-H., Kim, J.-K., Jang, H.-D. (2014). Genistein inhibits osteoclastic differentiation of RAW 264.7 cells via regulation of ROS production and scavenging. International Journal of Molecular Sciences, 15(6), 10605-10621.
Lephart, E. D. (2016). Skin aging and oxidative stress: Equol’s anti-aging effects via biochemical and molecular mechanisms. Ageing Research Reviews, 31, 36-54.
Li, R., Jia, Z., Trush, M. A. (2016). Defining ROS in biology and medicine. Reactive Oxygen Species (Apex, NC), 1(1), 9.
Lin, Y.-T., Mao, Y.-W., Imtiyaz, Z., Chiou, W.-F., Lee, M.-H. (2020). Comprehensive LC-MS/MS-based phytochemical perspectives and osteogenic effects of Uraria crinita. Food & Function, 11(6), 5420-5431.
Liu, X.-P., Cao, Y., Kong, H.-Y., Zhu, W.-F., Wang, G.-F., Zhang, J.-J., Qiu, Y.-C., Pang, J.-X. (2010). Antihyperglycemic and antihyperlipidemic effect of Uraria crinita water extract in diabetic mice induced by STZ and food. Journal of Medicinal Plants Research, 4(5), 370-374.
Liu, Y., Lu, T., Zhou, Z., Lu, Y., Zhang, Y. (2017). Study on feasibility of HaCaT epidermal model as an alternative to skin irritation in vitro. Chinese Journal of Reparative and Reconstructive Surgery, 31(10), 1262-1266.
Ma, Q. (2014). Advances in mechanisms of anti-oxidation. Discovery Medicine, 17(93), 121.
Mao, Y.-W., Lin, R.-D., Hung, H.-C., Lee, M.-H. (2014). Stimulation of osteogenic activity in human osteoblast cells by edible Uraria crinita. Journal of Agricultural and Food Chemistry, 62(24), 5581-5588.
McCallion, R., Po, A. L. W. (1993). Dry and photo–aged skin: and manifestations management. Journal of Clinical Pharmacy and Therapeutics, 18(1), 15-32.
McLafferty, E., Hendry, C., Farley, A. (2012). The integumentary system: anatomy, physiology and function of skin. Nursing Standard (through 2013), 27(3), 35.
Monteiro‐Riviere, N. A. (1997). Introduction to histological aspects of dermatotoxicology. Microscopy Research and Technique, 37(3), 171.
Morita, N., Arisawa, M., Nagase, M., Hsu, H., Chen, Y. (1977). Studies on the constituents of Formosan Leguminosae. II. The constituents in the leaves of Uraria crinita Desv (author's transl). Yakugaku zasshi: Journal of the Pharmaceutical Society of Japan, 97(6), 701-703.
Murakami, Y., Kawata, A., Ito, S., Katayama, T., Fujisawa, S. (2015). Radical-scavenging and anti-inflammatory activity of quercetin and related compounds and their combinations against RAW264. 7 cells stimulated with Porphyromonas gingivalis fimbriae. Relationships between anti-inflammatory activity and quantum chemical parameters. In Vivo, 29(6), 701-710.
Murray, J. C., Burch, J. A., Streilein, R. D., Iannacchione, M. A., Hall, R. P., Pinnell, S. R. (2008). A topical antioxidant solution containing vitamins C and E stabilized by ferulic acid provides protection for human skin against damage caused by ultraviolet irradiation. Journal of the American Academy of Dermatology, 59(3), 418-425.
Nimse, S. B., Pal, D. (2015). Free radicals, natural antioxidants, and their reaction mechanisms. RSC Advances, 5(35), 27986-28006.
Nusgens, B. V., Colige, A. C., Lambert, C. A., Lapière, C. M., Humbert, P., Rougier, A., Haftek, M., Richard A., Creidi, P. (2001). Topically applied vitamin C enhances the mRNA level of collagens I and III, their processing enzymes and tissue inhibitor of matrix metalloproteinase 1 in the human dermis. Journal of Investigative Dermatology, 116(6), 853-859.
Ohashi, H., Iokawa, Y. (2007). A Revision of Uraria (Leguminosae) in Taiwan. Taiwania, 52(2), 177-183.
Okawa, M., Akahoshi, R., Kawasaki, K., Nakano, D., Tsuchihashi, R., Kinjo, J., Nohara, T. (2019). Two new triterpene glycosides in the roots of Uraria crinita. Chemical and Pharmaceutical Bulletin, 67(2), 159-162.
Panche, A. N., Diwan, A. D., Chandra, S. R. (2016). Flavonoids: an overview. Journal of Nutritional Science, 5.
Poljšak, B., Dahmane, R. G., Godić, A. (2012). Intrinsic skin aging: The role of oxidative stress. Acta Dermatovenerol Alp Pannonica Adriat, 21(2), 33-36.
Prakash, D., Suri, S., Upadhyay, G., Singh, B. N. (2007). Total phenol, antioxidant and free radical scavenging activities of some medicinal plants. International Journal of Food Sciences and Nutrition, 58(1), 18-28.
Rahman, M. M., Gibbons, S., Gray, A. I. (2007). Isoflavanones from Uraria picta and their antimicrobial activity. Phytochemistry, 68(12), 1692-1697.
Rawlings, A. V., Matts, P. J. (2005). Stratum corneum moisturization at the molecular level: an update in relation to the dry skin cycle. Journal of Investigative Dermatology, 124(6), 1099-1110.
Rees, J. L. (2004). The genetics of sun sensitivity in humans. The American Journal of Human Genetics, 75(5), 739-751.
Schaefer, H., Stüttgen, G., Schalla, W., Gazith, J., Bauer, E. (1979). Principles of percutaneous absorption. In Toxicology (pp. 223-235): Elsevier.
Sen, S., De, B., Devanna, N., Chakraborty, R. (2013). Total phenolic, total flavonoid content, and antioxidant capacity of the leaves of Meyna spinosa Roxb., an Indian medicinal plant. Chinese Journal of Natural Medicines, 11(2), 149-157.
Si, H., Liu, D. (2007). Phytochemical genistein in the regulation of vascular function: new insights. Current Medicinal Chemistry, 14(24), 2581-2589.
Stiefel, C., Schwack, W. (2015). Photoprotection in changing times–UV filter efficacy and safety, sensitization processes and regulatory aspects. International Journal of Cosmetic Science, 37(1), 2-30.
Thien, D. D., Tai, B. H., Dai, T. D., Sa, N. H., Thuy, T. T., Hoang Anh, N. T., Tam, N. T. (2020). New phenolics from Uraria crinita (L.) DC. Natural Product Research, 1-8.
Tončić, R. J., Kezić, S., Hadžavdić, S. L., Marinović, B. (2018). Skin barrier and dry skin in the mature patient. Clinics in Dermatology, 36(2), 109-115.
Tu, P.-C., Chan, C.-J., Liu, Y.-C., Kuo, Y.-H., Lin, M.-K., Lee, M.-S. (2019). Bioactivity-guided fractionation and NMR-based identification of the immunomodulatory isoflavone from the roots of Uraria crinita (L.) Desv. ex DC. Foods, 8(11), 543.
van Acker, S. A., van Balen, G. P., van den Berg, D. J., Bast, A., van der Vijgh, W. J. (1998). Influence of iron chelation on the antioxidant activity of flavonoids. Biochemical Pharmacology, 56(8), 935-943.
Wang, Y., Zhang, X., Gong, L., Ruan, H., Pi, H., Zhang, Y. (2009). Studies on chemical constituents in roots of Uraria crinita. Chin. Pharm. J, 44, 1217-1220.
Williamson, E. M., Liu, X., Izzo, A. A. (2020). Trends in use, pharmacology, and clinical applications of emerging herbal nutraceuticals. British Journal of Pharmacology, 177(6), 1227-1240.
Wiseman, H. (2000). The therapeutic potential of phytoestrogens. Expert Opinion on Investigational Drugs, 9(8), 1829-1840.
Yao, L. H., Jiang, Y.-M., Shi, J., Tomas-Barberan, F., Datta, N., Singanusong, R., Chen, S. (2004). Flavonoids in food and their health benefits. Plant Foods for Human Nutrition, 59(3), 113-122.
Yen, G.-C., Lai, H.-H., Chou, H.-Y. (2001). Nitric oxide-scavenging and antioxidant effects of Uraria crinita root. Food Chemistry, 74(4), 471-478.
Zhang, J., Wider, B., Shang, H., Li, X., Ernst, E. (2012). Quality of herbal medicines: challenges and solutions. Complementary Therapies in Medicine, 20(1-2), 100-106.
Zhang, P., Omaye, S. T. (2000). β-Carotene and protein oxidation: effects of ascorbic acid and α-tocopherol. Toxicology, 146(1), 37-47.