臺灣博碩士論文加值系統

本研究之目的為探討11種樟科(Lauraceae)植物葉部抽出物之生物活性，評估抗真菌活性、抑制黃嘌呤氧化酶活性、抗氧化活性及抑制蕈類酪胺酸酶效果，並以斑馬魚為模式生物進行抑制黑色素生成活性的評估，期盼提升樟科植物天然物的用途。結果顯示11種樟科植物葉部抽出物不具有抑制黃嘌呤氧化酶活性，對於抗真菌的效果亦為有限。抗氧化活性分析結果發現，變葉新木薑子(Neolitsea aciculate)、小芽新木薑子(Neolitsea parvigemma)、鹿皮斑木薑子(Litsea coreana)與長葉木薑子(Litsea acuminata)葉部抽出物含有較多之總酚類化合物與總黃酮類化合物，在還原力、自由基清除活性與總抗氧化能力等評估皆具良好效果。
在蕈類酪胺酸酶評估中，以小芽新木薑子、鹿皮斑木薑子、內苳子(Lindera akoensis)及瓊楠(Beilschmiedia erythrophloia)等抽出物顯示最佳抑制活性，經酵素動力學分析，4種抽出物均屬競爭型抑制物。而在抑制斑馬魚黑色素生成效果的評估中，效果較佳的抽出物依序為長葉木薑子 (IC50 = 71.36 μg/mL) > 內苳子 (IC50 = 114.46 μg/mL) > 香桂(Cinnamomum subavenium) (IC50 = 117.32 μg/mL) > 五掌楠 (Neolitsea konishii) (IC50 = 122.19 μg/mL) > 樟樹(Cinnamomum camphora) (IC50 = 127.12 μg/mL) > 瓊楠 (IC50 = 188.16 μg/mL)，顯示長葉木薑子抽出物對於減少斑馬魚黑色素生成的表現最佳。綜合上述結果得知11種樟科植物中，變葉新木薑子、小芽新木薑子、鹿皮斑木薑子與長葉木薑子抽出物的抗氧化效果最佳，長葉木薑子抽出物的美白效果最好，未來可針對其活性成分加以探討，期盼能開發為良好之抗氧化劑及美白產品，增加林木天然物的利用價值。

The purpose of this study was to evaluate the bioactivities, including antifungal activity, xanthine oxidase inhibitory activity, antioxidant activity, antityrosinase activity, and melanogenesis inhibition activity in zebrafish (Danio rerio), of leaf extracts of 11 selected Lauraceae plants. Based on the results, 11 Lauraceae leaf extracts didn't possess the effect on antifungal and xanthine oxidase inhibitory activities. Leaf extracts of Neolitsea aciculate, Neolitsea parvigemma, Litsea coreana and Litsea acuminate contained the higher amount of total phenolic contents and total flavonoid contents, and these extract had superior antioxidant activities on reducing power, free radical scavenging activity, and total antioxidant capacity.
N. parvigemma extract, L. coreana extract, Lindera akoensis extract and B. erythrophloia extract exhibited the better mushroom tyrosinase inhibition activity. In antimelanogenesis assay, six leaf extracts possessed the higher melanogenesis inhibition activity in zebrafish with a decreasing order of L. acuminate > L. akoensis > Cinnamomum subavenium > Neolitsea konishii > C. camphora> Beilschmiedia erythrophloia, and L. acuminate leaf extract exhibited the highest melanin inhibitory activity in zebrafish. Accordingly, among 11 Lauraceae leaf extracts, N. aciculate extract, N. parvigemma extract, L. coreana extract, and L. acuminate extract shows the best antioxidant activity, and L. acuminate extract exhibited the best whitening effect, these extracts are worthy to do further investigation on their active constituents and with the potent to be antioxidant agents and skin whitening products.

摘要 I
Abstract III
目錄 IV
表目錄 VII
圖目錄 X
壹、前言 1
貳、文獻回顧 3
一、黑色素的形成 3
二、皮膚美白機制 5
(一)、抑制黑色素的形成 5
(二)、加速黑色素代謝 5
三、植物美白功效 6
四、樟科植物生物活性探討 12
(一)、抗細菌活性 12
(二)、抗真菌活性 14
(三)、抗氧化活性 15
(四)、抗發炎活性 16
(五)、抗癌活性 18
(六)、抑制黑色素生成 20
參、材料與方法 25
一、試驗材料 25
(一)、試材 25
(二)、黴菌種類 26
(三)、木材腐朽菌菌種 26
(四)、試驗藥品與溶劑 27
1. 試驗藥品 27
2. 溶劑 27
二、試驗方法 27
(一)、乙醇抽出物之萃取 27
(二)、抗真菌試驗 28
(三)、抑制黃嘌呤氧化酶活性測定 28
(四)、抗氧化活性試驗 28
1. 酚類化合物含量測定 28
2. 總黃酮類含量測定 29
3. 還原力測定 29
4. DPPH自由基清除能力之測定 30
5. 總抗氧化能力測定 30
(五)、酪胺酸酶試驗 31
(六)、酵素動力學 31
(七)、斑馬魚試驗 33
1. 斑馬魚培養條件 33
2. 斑馬魚胚胎美白能力評估 34
肆、結果與討論 35
一、樟科植物葉部抽出物抑制真菌活性效果之評估 35
(一)、樟科植物葉部抽出物抑制黴菌活性之評估 35
(二)、樟科植物葉部抽出物抑制木材腐朽菌之活性 39
二、樟科植物葉部抽出物抑制尿酸生成活性之評估 44
三、樟科植物葉部抽出物之抗氧化活性 45
(一)、樟科葉部抽出物之總酚類化合物含量 45
(二)、樟科葉部抽出物之總黃酮類含量 47
(三)、樟科葉部抽出物之還原能力 48
(四)、樟科葉部抽出物之DPPH自由基清除活性 50
(五)、樟科葉部抽出物之總抗氧化能力 54
四、樟科植物葉部抽出物對蕈類酪胺酸之抑制效果與酵素動力學探討 56
(一)、樟科植物葉部抽出物對蕈類酪胺酸酶抑制效果之評估 56
(二)、樟科植物葉部抽出物之酵素動力學探討 60
五、樟科植物葉部抽出物對斑馬魚黑色素的抑制活性 65
(一)、樟科植物葉部抽出物對斑馬魚黑色素生成的影響 65
(二)、樟科植物對斑馬魚黑色素之半數抑制濃度 68
伍、結論 74
陸、參考文獻 75

黃琪雅、馮苑琳、李文馨、張上鎮、張惠婷 (2012) 羅漢松屬植物抽出物之抗氧化 與抗酪胺酸酪活性評估。中華林學季刊453(3): 397-408。
溫國慶 (2007) 台灣產茜草科植物美白作用之篩選。中醫藥年報25(1): 239-256。
蔡蒼吾 (2010) 實驗動物管理與使用指南。行政院農業委員會。台北。第181- 183頁。
Apel, C., C. Geny, V. Dumontet, N. Birlirakis, F. Roussi, V. C. Pham, H. D. T. Mai, V. H. Nguyen, V. M. Chau and M. Litaudon (2014) Endiandric acid analogues from Beilschmiedia ferruginea as dual inhibitors of Bcl-xL/Bak and Mcl-1/Bid interactions. Journal of Natural Products 77: 1430-1437.
Chakraborty, A. K., Y. Funasaka, M. Komoto and M. Ichihashi (1998) Effect of arbutin on melanogenic proteins in human melanocytes. Pigment Cell Research 11: 206-212.
Chang, S. T., P. F. Chen and S. C. Chang (2001) Antibacterial activity of leaf essential oils and their constituents from Cinnamomum osmophloeum. Journal of Ethnopharmacology 77: 123-127.
Chang, T. S. (2009) An updated review of tyrosinase inhibitors. International Journal of Molecular Sciences 10: 2440-2475.
Chang, T.-S. (2012) Natural melanogenesis inhibitors acting through the down-regulation of tyrosinase activity. Materials 5: 1661-1685.
Cheng, M. J., I. L. Tsai, S. J. Lee, B. Jayaprakasam and I. S. Chen (2005) Steryl epoxide, secobutanolide and butanolides from the stem wood of Machilus zuihoensis. Phytochemistry 66: 1180-1185.
Chiang, H. M., H.W. Chen, Y.H. Huang, S.Y. Chan, C.C. Chen, W.C. Wu, and K. C. Wen (2012) Melanin: Biosynthesis Functions and Health Effects. Nova Science Publishers Inc. New York. pp. 1-76
Choi, T. Y., J. H. Kim, D. H. Ko, C. H. Kim, J. S. Hwang, S. Ahn, S. Y. Kim, C. D. Kim, J. H. Lee and T. J. Yoon (2007) Zebrafish as a new model for phenotype-based screening of melanogenic regulatory compounds. Pigment Cell Research 20: 120-127.
Chou, S. T., W. L. Chang, C. T. Chang, S. L. Hsu, Y. C. Lin and Y. Shih (2013) Cinnamomum cassia essential oil inhibits alpha-MSH-induced melanin production and oxidative stress in murine B16 melanoma cells. International Journal of Molecular Sciences 14: 19186-19201.
Dej-adisai, S., I. Meechai, J. Puripattanavong and S. Kummee (2014) Antityrosinase and antimicrobial activities from Thai medicinal plants. Archives of Pharmacal Research 37: 473-483.
Ebanks, J. P., R. R. Wickett and R. E. Boissy (2009) Mechanisms regulating skin pigmentation: The rise and fall of complexion coloration. International Journal of molecular sciences 10: 4066-4087.
Fanga, S. H., Y. K. Raob and Y. M. Tzengb (2004) Cytotoxic effect of trans-cinnamaldehyde from Cinnamomum osmophloeum leaves on human cancer cell lines. International Journal of Applied Science and Engineering 2: 136-147.
Goldsmith, P. (2004) Zebrafish as a pharmacological tool: the how, why and when. Current Opinion in Pharmacology 4: 504-512.
Ha, S. K., E. Moon, M. J. Lee, H. M. Park, E.-S. Yoo, M. S. Oh and S. Y. Kim (2009) Effect of the BuOH soluble fraction of Cinnamomum camphora on melanin biosynthesis. Korean Journal of Medicinal Crop Science 17: 293-300.
Ho, C. L., P. C. Liao, E. I. C. Wang and Y. C. Su (2011) Composition and antimicrobial activity of the leaf and twig oils of Litsea acutivena from Taiwan. Natural Product Communications 6: 1755-1758.
Huang, X. W., Y. C. Feng, Y. Huang and H. L. Li (2013) Potential cosmetic application of essential oil extracted from Litsea cubeba fruits from China. Journal of Essential Oil Research 25: 112-119.
John, A. J., V. P. Karunakaran and V. George (2007) Chemical composition and antibacterial activity of leaf oil of Neolitsea foliosa (Nees) gamble var. caesia (Meisner) gamble. Journal of Essential Oil Research 19: 498-500.
John, A. J., V. George, N. S. Pradeep and M. G. Sethuraman (2008) Chemical composition and antibacterial activity of the leaf, bark and fruit oils of Neolitsea fischeri gamble. Journal of Essential Oil Research 20: 279-282.
Kamakshi, R. (2012) Fairness via formulationsa review of cosmetic skin-lightening ingredients. Journal of Cosmetic Science 63: 43-54.
Kim, S. S., C. G. Hyun, Y. H. Choi and N. H. Lee (2013) Tyrosinase inhibitory activities of the compounds isolated from Neolitsea aciculata (Blume) Koidz. Journal of Enzyme Inhibition and Medicinal Chemistry 28: 685-689.
Kim, S. S., J. E. Kim, C. G. Hyun and N. H. Lee (2011) Neolitsea aciculata essential oil inhibits drug-resistant skin pathogen growth and propionibacterium acnes-induced inflammatory effects of human monocyte leukemia. Natural Product Communications 6: 1193-1198.
Kim, Y. J. and H. Uyama (2005) Tyrosinase inhibitors from natural and synthetic sources: structure, inhibition mechanism and perspective for the future. Cellular and Molecular Life Sciences 62: 1707-1723.
Kumar, K. J., J. C. Yang, F. H. Chu, S. T. Chang and S. Y. Wang (2010) Lucidone, a novel melanin inhibitor from the fruit of Lindera erythrocarpa Makino. Phytotherapy Research 24: 1158-1165.
Lee, H. J., E. A. Hyun, W. J. Yoon, B. H. Kim, M. H. Rhee, H. K. Kang, J. Y. Cho and E. S. Yoo (2006) In vitro anti-inflammatory and anti-oxidative effects of Cinnamomum camphora extracts. Journal of Ethnopharmacology 103: 208-216.
Lieschke, G. J. and P. D. Currie (2007) Animal models of human disease: zebrafish swim into view. Nature Reviews Genetics 8: 353-367.
Mapunya, M. B., R. V. Nikolova and N. Lall (2012) Melanogenesis and antityrosinase activity of selected South african plants. Evidence-Based Complementary and Alternative Medicine 2012: 374017.
Marongiu, B., A. Piras, S. Porcedda, E. Tuveri, E. Sanjust, M. Meli, F. Sollai, P. Zucca and A. Rescigno (2007) Supercritical CO(2) extract of Cinnamomum zeylanicum: Chemical characterization and antityrosinase activity. Journal of Agricultural and Food Chemistry 55: 10022-10027.
Matsuura, R., H. Ukeda, and M. Sawamura (2006) Tyrosinase inhibitory activity of citrus essential oils. Journal of Agricultural and Food Chemistry 54: 2309-2313.
Mendes, E., M. D. J. Perry and A. P. Francisco (2014) Design and discovery of mushroom tyrosinase inhibitors and their therapeutic applications. Expert Opinion on Drug Discovery 9: 533-554.
Mishra, S. K., P. N. Singha and S. D. Dubey (2013) Evaluation of antioxidant activities in ethanolic extract of Capparis zeylanica Linn. root. Notulae Scientia Biologicae 5(2): 127-132.


Nishaa, S., M. Vishnupriya, J. M. Sasikumar, P. C. Hephzibah and V. K. Gopalakrishnan (2012) Antioxdant activity of ethanolic extract of Maranta arundinacea .L tuberous rhizomes. Asian Journal of Pharmaceutical and Clinical Research 5(4): 85-88.
Oh, H. M., S. K. Choi, J. M. Lee, S. K. Lee, H. Y. Kim, D. C. Han, H. M. Kim, K. H. Son and B. M. Kwon (2005) Cyclopentenediones, inhibitors of farnesyl protein transferase and anti-tumor compounds, isolated from the fruit of Lindera erythrocarpa Makino. Bioorganic & Medicinal Chemistry 13: 6182-6187.
Perng, D. S., Y. H. Tsai, J. Cherng, J. S. Wang, K. S. Chou, C. W. Shih and J. M. Cherng (2016) Discovery of a novel anticancer agent with both anti-topoisomerase I and II activities in hepatocellular carcinoma SK-Hep-1 cells in vitro and in vivo: Cinnamomum verum component 2-methoxycinnamaldehyde. Journal of Drug Design, Development and Therapy 10: 141-153.
Salleh, W. M., F. Ahmad and K. H. Yen (2015) Chemical compositions and biological activities of the essential oils of Beilschmiedia madang Blume (Lauraceae). Archives of Pharmacal Research 38: 485-493.
Salleh, W. M. N. H. W., F. Ahmad, H. Y. Khong and R. Mohamed Zulkifli (2016) Comparative study of the essential oils of three Beilschmiedia species and their biological activities. International Journal of Food Science & Technology 51: 240-249.
Shan, B., Y. Z. Cai, J. D. Brooks and H. Corke (2007) Antibacterial properties and major bioactive components of cinnamon stick (Cinnamomum burmannii): Activity against foodborne pathogenic bacteria. Journal of Agricultural and Food Chemistry 55: 5484-5490.
Singh, G., S. Maurya, M. P. deLampasona and C. A. N. Catalan (2007) A comparison of chemical, antioxidant and antimicrobial studies of cinnamon leaf and bark volatile oils, oleoresins and their constituents. Food and Chemical Toxicology 45: 1650-1661.
Solano, F., S. Briganti, M. Picardo and G. Ghanem (2006) Hypopigmenting agents: an updated review on biological, chemical and clinical aspects. Pigment Cell Research 19: 550-571.
Venkatachalam, U and S. Muthukrishnan (2012) Free radical scavenging activity of ethanolic extract of Desmodium gangeticum. Journal of Acute Medicine 2(2): 36-42.
Wang, H. M., C. Y. Chen and Z. H. Wen (2011) Identifying melanogenesis inhibitors from Cinnamomum subavenium with in vitro and in vivo screening systems by targeting the human tyrosinase. Experimental Dermatology 20: 242-248.
Wang, N. and D. N. Hebert (2006) Tyrosinase maturation through the mammalian secretory pathway: bringing color to life. Pigment Cell Research 19: 3-18.
Wong, S. L., H. S. Chang, G. J. Wang, M. Y. Chiang, H. Y. Huang, C. H. Chen, S. C. Tsai, C. H. Lin and I. S. Chen (2011) Secondary metabolites from the roots of Neolitsea daibuensis and their anti-inflammatory activity. Journal of Natural Products 74: 2489-2496.
Yang, B. Y., L. Y. Kong, X. B. Wang, Y. M. Zhang, R. J. Li, M. H. Yang and J. G. Luo (2016) Nitric oxide inhibitory activity and absolute configurations of arylalkenyl alpha,beta-unsaturated delta/gamma-Lactones from Cryptocarya concinna. Journal of Natural Products 79: 196-203.
Yoon, W. J., J. Y. Moon, J. Y. Kang, G. O. Kim, N. H. Lee and C. G. Hyun (2010) Neolitsea sericea essential oil attenuates LPS-induced inflammation in RAW 264.7 macrophages by suppressing NF-kappa B and MAPK activation. Natural Product Communications 5: 1311-1316.
Zhang, C., C. Willett and T. Fremgen (2003) Zebrafish: an animal model for toxicological studies. Current Protocols in Toxicology 17: 1-18.