臺灣博碩士論文加值系統

過去有研究發現chrysin (5,7-dihydroxyflavone) 具有與芳香族酶抑制劑相似的作用，因此研究學者認為chrysin可能具有提高睪固酮濃度的功效。然而，過去關於補充chrysin的研究相當少數，且皆是以補充天然複方為主，並沒有補充單方chrysin的研究，因此本研究目的在探討補充市售單方chrysin是否影響睪固酮的合成，進而改變身體組成、肌力與運動表現，並藉由尿液檢測了解是否有違反運動禁藥規範之疑慮。受試對象為20位國立體育大學投擲隊隊員（男性14位；女性6位），實驗設計採配對設計 (matched pairs design)，依照競賽項目將受試者配對分組為 chrysin組（C組；20.9 ± 1.3歲）及placebo組（P組；20.7 ± 1.5歲）。受試者每餐飯後服用2顆chrysin膠囊或等量之安慰劑膠囊（3000mg/天），持續21天。於第1日、第8日、第22日早上進行血液、尿液樣本之採集。血液樣本檢測：睪固酮、游離睪固酮、雌二醇。尿液樣本檢測：睪固酮/表睪固酮比值（T/E比值）與睪固酮類緣物δ13C‰數值。統計方式以混合式二因子變異數分析 (two-way mixed design ANOVA) 檢驗C組及P組之各項數值是否具有組間或組內之差異性，並以LSD法進行事後比較各項數值以平均數 ± 標準差表示，顯著水準訂為 (p＜.05)。T/E比值與δ13C‰數值則以世界反運動禁藥機構 (World Anti-Doping Agency) 所訂定之檢測標準判斷是否違反運動禁藥規範。研究結果顯示各項指標並無發現組間之顯著差異 (p＞.05)。尿液檢測方面，T/E比值皆低於4，而δ13C‰數值分佈於正常範圍（>-28‰）。本研究結果顯示，連續服用21天chrysin並不會提升體內荷爾蒙的濃度，且無違反運動禁藥規範之疑慮。

In human, aromatase converts testosterone into estradiol. Chrysin has been shown as a potent aromatase inhibitor, which may increase the concentration of serum and urinary testosterone and lead to violate the doping control rule. However, the effects of the chrysin supplementation on blood and urinary testosterone levels in human have yet to be elucidated. PURPOSE: This study was to investigate whether the chrysin supplementation would increase serum testosterone by altering the synthesis of the testosterone pathway. Moreover, the ratio of testosterone to epitestosterone and the δ13C‰ value of testosterone metabolites in urine was measured. METHODS: Twenty throwing athletes of National Taiwan Sport University participated in this study. Subjects were randomly assigned in pairs (matched for athletic events) by use of a double blind design to receive chrysin (group C) or placebo (group P). All subjects were oral 3,000 mg of chrysin capsules or equal dosage of placebo per day for 21 days. Blood and urine samples were collected at day 1, 8, and 22 in the morning after overnight fasting. Blood samples were analyzed for serum testosterone, free testosterone, and estradiol. Urinary testosterone and epitestosterone was measured by GC/MS. A two-way mixed designed analysis of variance (ANOVA), with one within-subject factor (time trial), one between-subject factor (group C or P), and their interaction, was used to determine the overall 21-day treatment effect and level of significance. All values were expressed as the mean ± SD. Values of P<0.05 were considered significant. RESULTS: The data showed no significant difference in blood biomarkers between the groups C and P (testosterone: 4.7±2.3 vs. 4.3±1.6 ng/mL; free testosterone: 12.5±5.5 vs. 11.4± 3.5 pg/mL; estradiol: 43.2±22.1 vs. 49.6±18.7 pg/mL, p >.05). Urinary data of testosterone/epitestosterone ratio was less than 4. CONCLUSION: Our data demonstrated that chrysin supplementation for 21 days may not increase serum and urinary testosterone levels.

中文摘要 II
英文摘要 III
致 謝 詞 V
目 錄 VI
表 目 錄 VIII
圖 目 錄 IX
第壹章 緒 論 1
第一節 研究動機 1
第二節 研究目的 3
第貳章 文獻探討 4
第一節 Chrysin 的簡介與功效 4
第二節 睪固酮生成與代謝 8
第三節 T/E 值的檢測 10
第四節 同位素比值質譜儀的檢測 12
第五節 結 語 15
第參章 材料與方法 16
第一節 研究對象 16
第二節 實驗設計 17
第肆章 結 果 22
第一節 Chrysin對血液中荷爾蒙之影響 22
第二節 Chrysin 對 T/E 比值之影響 24
第三節 Chrysin 對睪固酮類緣物δ13C‰之影響 26
第四節 血液中之 chrysin 濃度 27
第伍章 討 論 30
第一節 Chrysin 對血液中荷爾蒙之影響 31
第二節 Chrysin 對T/E比值及睪固酮類緣物δ13C‰之影響 33
第三節 血液中之 Chrysin 濃度 34
第陸章 結論與建議 37
參考文獻 38
附錄一 補充chrysin對血液生化值之影響 47

中文部份
徐谷甫、許美智 (2007)。運動禁藥－芳香族酶抑制劑。大專體育，90，197-203。
許美智、詹貴惠 (2005)。運動藥物學。高雄市：春橋田股份有限公司。
黃懷玉 (2000)。補充冬蟲夏草配合重量訓練對於身體組成、血液生化值及最大總肌力之影響。國立體育學院運動科學研究所碩士論文，桃園縣。
劉雄、高建德 (2007)。黃岑研究進展。甘肅中醫學院學報，24(2)，46-51。
外文部份
Aguilera, R., Catlin, D. H., Becchi, M., Phillips, A., Wang, C., Swerdloff, R. S., Pope, H. G., &; Hatton, C. K. (1999). Screening urine for exogenous testosterone by isotope ratio mass spectrometric analysis of one pregnanediol and two androstanediols. Journal of Chromatography B: Biomedical Sciences and Applications, 727(1-2), 95-105.
Aguilera, R., Hatton, C. K., &; Catlin, D. H. (2002). Detection of epitestosterone doping by isotope ratio mass spectrometry. Clinical Chemistry, 48(4), 629-636.
Baume, N., Saudan, C., Desmarchelier, A., Strahm, E., Sottas, P. E., Bagutti, C., Cauderay, M., Schumacher, Y. O., Mangin, P., &; Saugy, M. (2006). Use of isotope ratio mass spectrometry to detect doping with oral testosterone undecanoate: inter-individual variability of 13C/12C ratio. Steroids, 71(5), 364-370.
Bourgogne, E., Herrou, V., Mathurin, J. C., Becchi, M., &; Ceaurriz, J. D. (2000). Detection of exogenous intake of natural corticosteroids by gas chromatography/combustion/isotope ratio mass spectrometry: application to misuse in sport. Rapid Communications in Mass Spectrometry, 14(24), 2343-2347.
Brown, G. A., Vukovich, M. D., Martini, E. R., Kohut, M. L., Franke, W. D., Jackson, D. A., &; King, D. S. (2001). Endocrine and lipid responses to chronic androstenediol-herbal supplementation in 30 to 58 year old men. Journal of the American College of Nutrition, 20(5), 520-528.
Catlin, D. H., Cowan, D. A., de la Torre, R., Donike, M., Fraisse, D., Oftebro, H., et al. (1996). Urinary testosterone (T) to epitestosterone (E) ratios by GC/MS. I. Initial comparison of uncorrected T/E in six international laboratories. Journal of Mass Spectrometry, 31(4), 397-402.
Ciftci, O., Ozdemir, I., Aydin, M., &; Beytur, A. (2012). Beneficial effects of chrysin on the reproductive system of adult male rats. Andrologia, 44, 181-186.
Cowan, D. A., Kicman, A. T., Walker, C. J., &; Wheeler, M. J. (1991). Effect of administration of human chorionic gonadotrophin on criteria used to assess testosterone administration in athletes. The Journal of Endocrinology, 131(1), 147-154.
Critchfield, J. W,, Butera, S. T., &; Folks, T. M. (1996). Inhibition of HIV activation in latently infected cells by flavonoid compounds. AIDS Research and Human Retroviruses, 12, 39-46.
Dalzell, D. P., &; El Attar, T. M. (1973). Gas chromatographic determination of urinary excretion of testosterone, epitestosterone, and androstenedione in pre-adolescent and adolescent children. The Journal of Clinical Endocrinology and Metabolism, 36, 1237-1243.
Dray, F., &; Ledru, M. J. (1966). Metabolism of epitestosterone. Absence of peripheral interconversion of epitestosterone and testosterone and existence of a production of epitestosterone sulfate in normal adult men. Comptes Rendus Hebdomadaires des Séances de L'Académie des Sciences. Série D: Sciences Naturelles, 262, 679-681.
Falk, O., Palonek, E., &; Bjorkhem, I. (1988). Effect of ethanol on the ratio between testosterone and epitestosterone in urine. Clinical Chemistry, 34, 1462-1464.
Ferchaud, V., Bizec, B. L., Monteau, F., &; Andre, F. (2000). Characterization of exogenous testosterone in livestock by gas chromatography/combustion/isotope ratio mass spectrometry: influence of feeding and age. Rapid Communications in Mass Spectrometry, 14(8), 652-656.
Funderburgh, L. J., Zipf, W. B., &; Sotos, J. F. (1983). Direct measurement of testosterone in a pediatric center, with use of a radioimmunoassay kit and unextracted serum. Clinical Chemistry, 29, 1796-1798.
Galijatovic, A., Otake, Y., Walle, U. K., &; Walle, T. (1999). Extensive metabolism of the flavonoid chrysin by human Caco-2 and Hep G2 cells. Xenobiotica, 29(12), 1241-1256.
Gambelunghe, C., Rossi, R., Sommavilla, M., Ferranti, C., Rossi, R., Ciculi, C., Gizzi, S., Micheletti, A., &; Rufini, S. (2003). Effects of chrysin on urinary testosterone levels in human males. Journal of Medicinal Food, 6(4):387-90.
Gardana, C., Simonetti, P., Berti, C., &; Pietta, P. (2007). Evaluation of propolis polyphenols absorption in humans by liquid chromatography/tandem mass spectrometry. Rapid Communications in Mass Spectrometry, 21, 3849-3854.
Gryglewski, R. J., Korbut, R., Robak, J., &; Swies, J. (1987). On the mechanism of antithrombotic action of flavonoids. Biochemical Pharmacology, 36, 317-322.
Havsteen, B. (1983). Falvonoids, a class of natural products of high pharmacological potency. Biochemical Pharmacology, 32, 1141-1148.
Hertog, M. G. L., Feskens, E. J. M., Hollman, P. C. H., Katan, M. B., &; Kromhout, D. (1993). Dietary antioxidant flavonoids and risk of coronary heart disease. Lancet, 342, 1007-1011.
Ibrahim, E., &; Abul-Hajj, Y. J. (1990). Aromatase inhibition by flavonoids. Journal of Steroid Biochemistry and Molecular Biology, 37(2), 257-260.
Jeong, H. J., Shin, Y. G., Kim, I. H., &; Pezzuto, J. M. (1999). Inhibition of aromatase activity by flavonoids. Archives of Pharmacal Research, 22(3), 309-312.
Keli, S. O., Hertog, M. G. L., Feskens, E. J. M., &; Kromhout, D. (1996). Dietary flavonoids, antioxidant vitamins, and incidence of stroke: The Zutphen study. Archives of Internal Medicine, 156, 637-642.
Kellis, J. T. Jr., &; Vickery, L. E. (1984). Inhibition of human estrogen synthetase (aromatase) by flavones. Science, 225(4666), 1032-1034.
Korenman, S. G., Wilson, H., &; Lipsett, M. B. (1964). Isolation of 17-alpha-hydroxyandrost-4-en-3-one (epitestosterone) from human urine. The Journal of Biological Chemistry, 239, 1004-1006.
Lapcik, O., Hampl, R., Hill, M., &; Starka, L. (1995). Plasma levels of epitestosterone from prepuberty to adult life. The Journal of Steroid Biochemistry and Molecular Biology, 55,405-408.
Leinonen, A., Kuuranne, T., Kotiaho, T., &; Kostiainen, R. (2004). Screening of free 17-alkyl-substituted anabolic steroids in human urine by liquid chromatography–electrospray ionization tandem mass spectrometry. Steroids, 69, 101–109.
Le Marchand, L., Murphy, S. P., Hankin, J. H., Wilkens, L. R., &; Kolonel, L. N. (2000). Intake of flavonoids and lung cancer. Journal of the National Cancer Institute, 92, 154-160.
Linnet, K. (1993). Effect of the biological matrix on the urinary testosterone/epitestosterone ratio measured by gas chromatography/mass spectrometry in doping analysis. Biological Mass Spectrometry, 22(7), 412-418.
Masse, R., Ayotte, C., &; Dugal, R. (1989). Integrated methodological approach to the gas chromatographic-mass spectrometric analysis of anabolic steroid metabolites in urine. Journal of Chromatography, 489, 23–50.
Mauras, N., Lima, J., Patel, D., Rini, A., di Salle, E., Kwok, A., &; Lippe, B. (2003). Pharmacokinetics and dose finding of a potent aromatase inhibitor, aromasin (Exemestane), in young males. Journal of Clinical Endocrinology and Metabolism, 88(12), 5951-5956.
Mohammed, H. A., Ba, L. A., Burkholz, T., Schumann, E., Diesel, B., Zapp, J., Kiemer, A. K., Ries, C., Hartmann, R. W., Hosny, M., &; Jacob, C. (2011). Facile synthesis of chrysin-derivatives with promising activities as aromatase inhibitors. Natural Product Communications, 6(1), 31-34.
Muller, L., &; Phillipou, G. (1987). Quantification of 5 alpha- and 5 beta-androstanediols in urine by gas chromatography-mass spectrometry. Clinical Chemistry, 33(2), 256-260.
Navajas, R., Imaz, C., Carreras, D., Garcia, M., Perez, M., Rodriguez, C., Rodriguez, A. F., &; Cortes, R. (1995). Determination of epitestosterone and testosterone in urine by high-performance liquid chromatography. Journal of Chromatography. B, Biomedical Applications, 673(2), 159-164.
Nier, A. O. (1950). A redetermination of the relative abundances of the isotope of carbon, nitrogen, oxygen, argon and potassium. Physical Review, 77, 789-793.
Pope, H.G., Katz, D.L. &; Champoux, R. (1988). Anabolic-androgenic steroid use among 1010 college men. Physician and Sports Medicine, 16, 75-81.
Rogozkin, V. (1979). Metabolic effects of anabolic steroid on skeletal muscle. Medicine and Science in Sports, 11(2), 160-163
Rosland, M. C., &; Mahmoud, A. M. (2011). Hormones and prostate carcinogenesis: Androgens and estrogens. Journal of Carcinogenesis, 2011; 10, 33.
Saudan, C., Baume, N., Mangin, P., &; Saugy, M. (2004). Urinary analysis of 16(5alpha)-androsten-3alpha-ol by gas chromatography/combustion/isotope ratio mass spectrometry: implications in anti-doping analysis. Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences, 810(1), 157-164.
Saudan, C., Kamber, M., Barbati, G., Robinson, N., Desmarchelier, A., &; Mangin, P. (2006). Longitudinal profiling of urinary steroids by gas chromatography/combustion/isotope ratio mass spectrometry: diet change may result in carbon isotopic variations. Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences, 831(1-2), 324-327.
Segura, J., Pascual, J. A., Ventura, R., Ustaran, J. I., Cuevas, A., &; Gonzalez, R. (1993). International cooperation in analytical chemistry: experience of antidoping control at the XI Pan American Games. Clinical Chemistry, 39(5), 836-845.
Shackleton, C. H., Roitman, E., Phillips, A., &; Chang, T. (1997). Androstanediol and 5-androstenediol profiling for detecting exogenously administered dihydrotestosterone, epitestosterone, and dehydroepiandrosterone: potential use in gas chromatography isotope ratio mass spectrometry. Steroids, 62(10), 665-673.
Shen, M., Chan, T. H., &; Dou, Q. P. (2012). Targeting tumor ubiquitin-proteasome pathway with polyphenols for chemosensatization. Anti-cancer Agents in Medicinal Chemistry, 5(1), 11-16.
Siess, M. H., Le Bon, A. M., Canivenc –Lavier, M. C., Amiot, M. J., Sabatier, S., Aubert, S. Y., &; Suschetet, M. (1996). Flavonoids of honey and propolis: Characterization and effects on hepatic drug-metabolizing enzymes and benzo[α]pyrene- DNA binding in rats. Journal of Agricultural and Food Chemistry, 44, 2297-2301
Ueki, M., &; Okano, M. (1999). Analysis of exogenous dehydroepiandrosterone excretion in urine by gas chromatography/combustion/isotope ratio mass spectrometry. Rapid Communications in Mass Spectrometry, 13(22), 2237-2243.
van Meeuwen, J. A., Korthagen, N., de Jong, P. C., Piersma, A. H., &; van den Berg, M. (2007). (Anti)estrogenic effects of phytochemicals on human primary mammary fibroblasts, MCF-7 cells and their co-culture. Toxicology and Applied Pharmacology, 221(3), 372-383.
Van Thuyne, W., &; Delbeke, F. T. (2005). Validation of a GC/MS screening method for anabolizing agents in aqueous nutritional supplements. Journal of Chromatographic Science, 43(1), 2-6.
Venturelli, E., Manzari, A., Cavalleri, A., Benzo, M., Secreto, G., &; Marubini, E. (1992). Urinary testosterone measurement by gas chromatography after solid-phase extraction and high-performance liquid chromatography. Journal of Chromatography, 582(1-2), 7-12.
Walle, T., Otake, Y., Brubaker, J. A., Walle, U. K., &; Halushka, P. V. (2001). Disposition and metabolism of the flavonoid chrysin in normal volunteers. British Journal of Clinical Pharmacology, 51, 143-146.
Wang, C., Catlin, D. H., Starcevic, B., Heber, D., Ambler, C., Berman, N., Lucas, G., Leung, A., Schramm, K., Lee, P. W. N., Hull, L., &; Swerdloff, R. S. (2005). Low-fat high-fiber diet decreased serum and urine androgens in men. Journal of Clinical Endocrinology and Metabolism, 90(6), 3550-3559.
Wang, X., &; Morris, M. E. (2007). Effects of the flavonoid chrysin on nitrofurantoin pharmacokinetics in rats: potential involvement of ABCG2. Drug Metabolism and Disposition, 35, 268-274.
Weiss, L. W., Cureton, K. J., &; Thompson, F.N. (1983). Comparison of serum testosterone and androstenedione responses to weight lifting in men and women. European Journal of Applied Physiology and Occupational Physiology, 50(3), 413-419.
Wilson, J. D. (1988). Androgen abuse by athletes. Endocrine Reviews, 9(2), 181-199.
World Anti-Doping Agency. (2007). Prohibited List 2007. Retreived from http://www.wada-ama.org/rtecontent/document/2007_List_En.pdf
Yamada, K., &; Aizawa, Y. (1983). A simple method for testosterone measurement by high-performance liquid chromatography-UV spectrophotometer. Journal of Pharmacological Methods, 10(3), 167-173.