肥胖和不孕症已是開發中國家常見之文明病。而肥胖常會伴隨許多高血脂和糖尿病等代謝症候群等發生，主要因為氧化壓力使得體內產生慢性發炎，導致精子功能低下、睪丸損傷和性腺荷爾蒙分泌紊亂，進而造成男性不孕。紫錐花 (Echinacea purpurea) 已被證實具有抗氧化及抗發炎的活性，故本研究目的以紫錐花乙醇萃取物 (Echinacea purpurea ethanol extract, EPE) 探討高脂飲食 (high fat diet, HFD) 誘發肥胖所產生高血脂、慢性發炎和氧化壓力造成雄性生殖功能損害之保護功效。動物實驗以 Sprague-Dawley (SD) 雄性大鼠給予 5 週 45 % HFD 誘導肥胖模式後，管餵 EPE 劑量 93、186、465 mg/kg，並持續 5 週 HFD，結果顯示 EPE 隨劑量增加能有效改善體重、降低血清及肝臟中的三酸甘油酯 (TG) 並提升高密度脂蛋白 (HDL-C) 含量、降低空腹血糖含量、血清中睪固酮含量有上升的趨勢；此外，提升肝臟中磷酸化 AMPK 和 PPAR-α，並抑制 PPAR-γ 的蛋白表現，能促進脂質代謝中脂肪酸氧化及抑制脂肪形成；睪丸中 StAR 和 17β-HSD3 上升，能提升睪固酮合成。血清及精子中一氧化氮 (NO) 及過氧化脂質 (MDA) 含量下降，提高血清、肝臟及睪丸中的抗氧化酶活性，包括超氧歧化酶 (SOD)、麩胱甘肽過氧化酶 (GPx)、麩胱甘肽 (GSH)、過氧化氫酶 (Cat)，降低睪丸內促發炎激素 TNF-α 和 IL-6，體內中抗氧化酶增加及促發炎激素下降，能降低肥胖所帶來的氧化壓力傷害。生殖參數上，肥胖對精子所造成的傷害，其精子總數、泳動率、型態、粒線體膜電位和 DNA 損傷狀況皆有明顯改善。總結，給予紫錐花乙醇萃取物，對於肥胖大鼠，具有調解血脂、體內抗氧化和抗發炎，並進一步改善因肥胖所造成的生殖系統傷害。

Obesity’s risk appears to originate from disruption in adipose tissue function leading to a chronic inflammatory state. It comes up with various disorders such as type 2 diabetes, hyperlipidemia, coronary diseases, and also infertility. Obesity is rapidly becoming a worldwide epidemic. Some studies have shown a relationship between obesity and infertility. Echinacea purpurea has long been used as a traditional herb for immune-enhancing, and is also demonstrated to have anti-inflammatory and anti-oxidant effects. But male obesity induced chronic inflammatory and oxidative stress on reproductive damage of this material has not yet been investigated. Thus, the aim of the present study focuses on that if E. purpurea has potential to prevent high fat diet-induced hyperlipidemia and reproductive damage. In vivo, male sprague-dawley rats were fed high fat diet for 5 weeks to induce obesity. EPE was tested in three doses (93, 186, 465 mg/kg per day) during 5 weeks high fat diet to determine whether E. purpurea reduces high fat diet-induced hyperlipidemia and its reproductive dysfunction. The results showed that EPE decreased body weight, fat mass, TG, fasting blood glucose index and increase of HDL-C level in high dose EPE group. Next, this study found that the level of antioxidant enzymes were increased in high dose EPE group; lipid peroxidation and the level of inflammation cytokines including NO, IL-6 and TNF-α were decreased. The sperm parameters such as counts, mobility, morphology, DNA damage and mitochondrial membrane potential were all ameliorated. Besides, the protein expression of PPAR-α, PPAR-γ, AMPKα1/2 phosphorylation in liver tissue, as well as StAR and 17β-HSD3 in testis were raised, showing increase of lipid metabolism and testosterone synthesis under the effect of EPE, respectively. In conclusion five weeks administration of E. purpurea beneficially regulated hyperlipidemia, anti-oxidant activity, anti-inflammation and reproductive parameters in obesity rats.

摘要 I
Abstract II
目錄 III
圖目錄 V
表目錄 VI
附錄目錄 VII
名詞縮寫表 VIII
壹、緒論 1
貳、文獻回顧 2
一、肥胖與代謝症候群 2
1.1 胰島素 (Insulin) 2
1.2 瘦素 (Leptin) 3
二、男性生殖與不孕症 3
2.1 男性生殖系統 3
2.2 男性不孕症 3
三、肥胖和男性生殖之關係 4
3.1 生殖內分泌 4
3.2 氧化壓力 5
四、紫錐花 6
4.1 紫錐花之簡介 6
4.2 紫錐花之成分 6
4.3 紫錐花之生理活性 7
叁、實驗設計 8
一、實驗設計 8
1.1 動物實驗設計 8
1.2 犧牲後分析項目 9
肆、實驗材料 10
一、材料與樣品 10
1.1 材料來源 10
1.2 樣品製備 10
1.3 實驗動物 10
1.4 動物飼料 10
二、藥品材料 11
三、儀器設備 12
伍、實驗方法 13
一、動物試驗 13
1.1 肥胖大鼠誘導與分組 13
1.2 實驗動物飼養 13
1.3 分析項目 14
陸、統計分析 25
柒、實驗結果 26
一、EPE 對於肥胖大鼠體重、臟器重量、攝食量和脂肪含量之分析 26
二、EPE 對於肥胖大鼠血漿生化指標之影響 26
2.1 血漿中血糖與血脂含量變化評估 26
2.2 血漿丙氨酸轉氨酶、天門冬氨酸轉氨酶、尿素及肌酸酐含量變化 26
2.3 血漿中睪固酮含量 26
三、EPE 對於肥胖大鼠體內抗氧化及抗發炎之改善情形 27
四、EPE 對於肥胖脂質代謝之改善情形 27
五、EPE 對於肥胖病大鼠生殖功能之探討 27
5.1 睪丸和副睪重量 27
5.2 睪丸組織型態 27
5.3 睪固酮合成路徑之探討 28
5.4 精子參數 28
5.5 精子粒線體膜電位、DNA 損傷 28
捌、討論 29
一、EPE 對於肥胖大鼠體重、BMI 指數和攝食量之分析 29
二、EPE 對於肥胖大鼠臟器和脂肪含量之分析 29
三、EPE 對於肥胖大鼠血漿生化指標之影響 29
3.1 血漿中血糖與血脂含量變化評估 29
3.2 血漿丙氨酸轉氨酶、天門冬氨酸轉氨酶、尿素及肌酸酐含量變化 29
四、EPE 對於肥胖大鼠體內抗氧化及抗發炎之改善情形 30
五、EPE 對於肥胖脂質代謝之改善情形 30
六、EPE 對於肥胖病大鼠生殖功能之探討 31
6.1 睪丸、副睪重量 31
6.2 睪丸組織型態 31
6.3 睪固酮合成路徑之探討 31
6.4 精子參數之影響 31
6.5 精子膜電位與 DNA 損傷 32
玖、結論 33
拾、文獻 34
拾壹、圖表 40
拾貳、附錄 68

什、文獻
行政院衛生福利部 (2015) 國民健康署。臺北。臺灣。
行政院衛生福利部 (2013) 國民營養健康狀況變遷調查。臺北。臺灣。
行政院衛生福利部 (2007) 成人代謝症候群之判定標準。臺北。臺灣。
甘邱涵。2012。膳食補充椴木牛樟芝子實體對高脂飲食誘發肥胖雄性大鼠生殖功能之影響。國立臺灣海洋大學食品科學系碩士論文。基隆。臺灣。
Abei H. 1984. Catalase in vitro. Methods in Enzymology 105:121-126
Ahima RS, Prabakaran D, Mantzoros C, Qu D, Lowell B, Maratos-Flier E, Flier JS. 1996. Role of leptin in the neuroendocrine response to fasting. Nature 382:250-252
Baazeem A, Belzile E, Ciampi A, Dohle G, Jarvi K, Salonia A, Weidner W, Zini A. 2011. Varicocele and male factor infertility treatment: a new meta-analysis and review of the role of varicocele repair. European Urology 60(4):796-808.
Bakos H, Mitchell M, Setchell BP, Lane M. 2011. The effect of paternal diet‐induced obesity on sperm function and fertilization in a mouse model. International Journal of Andrology 34(5pt1):402-410.
Balandrin MF, Kinghorn AD, Farnsworth NR. 1993. Plant-derived natural products in drug discovery and development: an overview. ACS Symposium Series 534:2-12.
Barnes J, Anderson LA, Gibbons S, Phillipson JD. 2005. Echinacea species (Echinacea angustifolia (DC.) Hell., Echinacea pallida (Nutt.) Nutt., Echinacea purpurea (L.) Moench): a review of their chemistry, pharmacology and clinical properties. Journal of Pharmacy and Pharmacology 57(8):929-954.
Bauer R, Remiger P, Wagner H. 1989. Alkamides from the roots of Echinacea angustifolia. Phytochemistry 28(2):505-508.
Baumber J, Ball BA, Gravance CG, Medina V, DAVIES‐MOREL MC. 2000. The effect of reactive oxygen species on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential, and membrane lipid peroxidation. Journal of Andrology 21(6):895-902.
Binns S, Hudson J, Merali S, Arnason J. 2002. Antiviral activity of characterized extracts from echinacea spp.(Heliantheae: Asteraceae) against herpes simplex virus (HSV-I). Planta Medica 68(9):780-783.
Binns S, Purgina B, Bergeron C, Smith M, Ball L, Baum B, Arnason J. 2000. Light-mediated antifungal activity of Echinacea extracts. Planta Medica 66(3):241-244.
Boivin J, Bunting L, Collins JA, Nygren KG. 2007. International estimates of infertility prevalence and treatment-seeking: potential need and demand for infertility medical care. Human Reproduction 22(6):1506-1512.
Chiu K, Chuang S, Sung J. 2006. Both anti-oxidation and lipid-carbohydrate conversion enhancements are involved in priming-improved emergence of Echinacea purpurea seeds that differ in size. Scientia Horticulturae 108(2):220-226.
Choi MY, Song HS, Hur HS, Sim SS. 2008. Whitening activity of luteolin related to the inhibition of cAMP pathway in α-MSH-stimulated B16 melanoma cells. Archives of Pharmacal Research 31(9):1166-1171.
Clifford L, Nair M, Rana J, Dewitt D. 2002. Bioactivity of alkamides isolated from Echinacea purpurea (L.) Moench. Phytomedicine 9(3):249-253.
Condorelli R, Calogero A, Vicari E, La Vignera S. 2014. Chronic consumption of alcohol and sperm parameters: our experience and the main evidences. Andrologia 47(4):368-379.
de La Serre CB, Ellis CL, Lee J, Hartman AL, Rutledge JC, Raybould HE. 2010. Propensity to high-fat diet-induced obesity in rats is associated with changes in the gut microbiota and gut inflammation. American Journal of Physiology-Gastrointestinal and Liver Physiology 299(2):G440-G448.
Delarue J, Magnan C. 2007. Free fatty acids and insulin resistance. Current Opinion in Clinical Nutrition &; Metabolic Care 10(2):142-148.
Desai NR, Kesari KK, Agarwal A. 2009. Pathophysiology of cell phone radiation: oxidative stress and carcinogenesis with focus on male reproductive system. Reprod Biol Endocrinol 7(114):1-9.
Dixit VD. 2008. Adipose-immune interactions during obesity and caloric restriction: reciprocal mechanisms regulating immunity and health span. Journal of Leukocyte Biology 84(4):882-892.
El-Haschimi K, Pierroz DD, Hileman SM, Bj#westeur057#rb#westeur039#k C, Flier JS. 2000. Two defects contribute to hypothalamic leptin resistance in mice with diet-induced obesity. Journal of Clinical Investigation 105(12):1827.
Elshal MF, El-Sayed IH, Elsaied MA, El-Masry SA, Kumosani TA. 2009. Sperm head defects and disturbances in spermatozoal chromatin and DNA integrities in idiopathic infertile subjects: association with cigarette smoking. Clinical Biochemistry 42(7):589-594.
Esteves SC, Agarwal A. 2011. Novel concepts in male infertility. International Braz J Urol 37(1):5-15.
Fisher JS. 2004. Environmental anti-androgens and male reproductive health: focus on phthalates and testicular dysgenesis syndrome. Reproduction 127(3):305-315.
Fluck D, Valentine P, Treister B, Higgs B, Reid D, Steiner R, Mounsey J. 1967. Right heart pressures in acute myocardial infarction. British Heart Journal 29(5):748.
Fujii J, Iuchi Y, Matsuki S, Ishii T. 2003. Cooperative function of antioxidant and redox systems against oxidative stress in male reproductive tissues. Asian Journal of Andrology 5(3):231-242.
Galimova E, Amirova Z, Galimov SN. 2014. Dioxins in the semen of men with infertility. Environmental Science and Pollution Research:1-4.
Garolla A, Torino M, Sartini B, Cosci I, Patassini C, Carraro U, Foresta C. 2013. Seminal and molecular evidence that sauna exposure affects human spermatogenesis. Human Reproduction 28(4):877-885.
Gharagozloo P, Aitken RJ. 2011. The role of sperm oxidative stress in male infertility and the significance of oral antioxidant therapy. Human Reproduction 26(7):1628-1640.
Ghibaudi L, Cook J, Farley C, Heek M, Hwa JJ. 2002. Fat Intake Affects Adiposity, Comorbidity Factors, and Energy Metabolism of Sprague‐Dawley Rats. Obesity Research 10(9):956-963.
Goel V, Chang C, Slama JV, Barton R, Bauer R, Gahler R, Basu TK. 2002. Alkylamides of Echinacea purpurea stimulate alveolar macrophage function in normal rats. International Immunopharmacology 2(2):381-387.
Hammoud AO, Wilde N, Gibson M, Parks A, Carrell DT, Meikle AW. 2008. Male obesity and alteration in sperm parameters. Fertility and Sterility 90(6):2222-2225.
Harris ID, Fronczak C, Roth L, Meacham RB. 2011a. Fertility and the aging male. Reviews in Urology 13(4):e184-e190.
Harris RM, Weiss J, Jameson JL. 2011b. Male hypogonadism and germ cell loss caused by a mutation in Polo-like kinase 4. Endocrinology 152(10):3975-3985.
Harton GL, Tempest HG. 2012. Chromosomal disorders and male infertility. Asian J Androl 14(1):32-39.
Hwang JT, Lee MS, Kim HJ, Sung MJ, Kim HY, Kim MS, Kwon DY. 2009. Antiobesity effect of ginsenoside Rg3 involves the AMPK and PPAR‐γ signal pathways. Phytotherapy Research 23(2):262-266.
James PT, Leach R, Kalamara E, Shayeghi M. 2001. The worldwide obesity epidemic. Obesity Research 9(S11):228S-233S.
Jones N, HARRISON GA. 1956. Genetically determined obesity and sterility in the mouse. Proceedings. Society for the Study of Fertility 9:51-64.
Kelly DM, Jones TH. 2013. Testosterone: a metabolic hormone in health and disease. Journal of Endocrinology 217(3):R25-R45.
Kinghorn AD, Balandrin MF. 1993. Human medicinal agents from plants. American Chemical Society.
Luettig B, Steinm#westeur061#ller C, Gifford G, Wagner H, Lohmann-Matthes M-L. 1989. Macrophage activation by the polysaccharide arabinogalactan isolated from plant cell cultures of Echinacea purpurea. Journal of the National Cancer Institute 81(9):669-675.
Magnusdottir EV, Thorsteinsson T, Thorsteinsdottir S, Heimisdottir M, Olafsdottir K. 2005. Persistent organochlorines, sedentary occupation, obesity and human male subfertility. Human Reproduction 20(1):208-215.
Mah P, Wittert G. 2010. Obesity and testicular function. Molecular and Cellular Endocrinology 316(2):180-186.
Marklund S, Marklund G. 1974. Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. European Journal of Biochemistry 47(3):469-474.
McDougall B, King PJ, Wu BW, Hostomsky Z, Reinecke MG, Robinson WE. 1998. Dicaffeoylquinic and dicaffeoyltartaric acids are selective inhibitors of human immunodeficiency virus type 1 integrase. Antimicrobial Agents and Chemotherapy 42(1):140-146.
Mishima S, Saito K, Maruyama H, Inoue M, Yamashita T, Ishida T, Gu Y. 2004. Antioxidant and immuno-enhancing effects of Echinacea purpurea. Biological and Pharmaceutical Bulletin 27(7):1004-1009.
Mitchell M, Bakos HW, Lane M. 2011. Paternal diet-induced obesity impairs embryo development and implantation in the mouse. Fertility and Sterility 95(4):1349-53.
Myers MG, Cowley MA, M#westeur061#nzberg H. 2008. Mechanisms of leptin action and leptin resistance. Annu. Rev. Physiol. 70:537-556.
Novelli E, Diniz Y, Galhardi C, Ebaid G, Rodrigues H, Mani F, Fernandes A, Cicogna A, Novelli Filho J. 2007. Anthropometrical parameters and markers of obesity in rats. Laboratory Animals 41(1):111-119.
O'Flaherty C, de Lamirande E, Gagnon C. 2005. Reactive oxygen species and protein kinases modulate the level of phospho-MEK-like proteins during human sperm capacitation. Biology of Reproduction 73(1):94-105.
Palmeira CM, Moreno AJ, Madeira VM, Wallace KB. 1996. Continuous monitoring of mitochondrial membrane potential in hepatocyte cell suspensions. Journal of Pharmacological and Toxicological Methods 35(1):35-43.
Palmer NO, Bakos HW, Fullston T, Lane M. 2012. Impact of obesity on male fertility, sperm function and molecular composition. Spermatogenesis 2(4):253-263.
Pascot A, Lemieux I, Prud'homme D, Tremblay A, Nadeau A, Couillard C, Bergeron J, Lamarche B, Despr#westeur042#s J-P. 2001. Reduced HDL particle size as an additional feature of the atherogenic dyslipidemia of abdominal obesity. Journal of Lipid Research 42(12):2007-2014.
Pavlovich CP, King P, Goldstein M, Schlegel PN. 2001. Evidence of a treatable endocrinopathy in infertile men. The Journal of Urology 165(3):837-841.
Placer ZA, Cushman LL, Johnson BC. 1966. Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Analytical Biochemistry 16(2):359-364.
Raso GM, Pacilio M, Carlo G, Esposito E, Pinto L, Meli R. 2002. In‐vivo and in‐vitro anti‐inflammatory effect of Echinacea purpurea and Hypericum perforatum. Journal of Pharmacy and Pharmacology 54(10):1379-1383.
Reagan-Shaw S, Nihal M, Ahmad N. 2008. Dose translation from animal to human studies revisited. The FASEB Journal 22(3):659-661.
Robinson WE. 1998. L-chicoric acid, an inhibitor of human immunodeficiency virus type 1 (HIV-1) integrase, improves on the in vitro anti-HIV-1 effect of Zidovudine plus a protease inhibitor (AG1350). Antiviral Research 39(2):101-111.
Roesler J, Steinm#westeur061#ller C, Kiderlen A, Emmend#westeur055#rffer A, Wagner H, Lohmann-Matthes M-L. 1991. Application of purified polysaccharides from cell cultures of the plant Echinacea purpurea to mice mediates protection against systemic infections with Listeria monocytogenes and Candida albicans. International Journal of Immunopharmacology 13(1):27-37.
Samuel VT, Shulman GI. 2012. Mechanisms for insulin resistance: common threads and missing links. Cell 148(5):852-871.
Schmid TE, Eskenazi B, Marchetti F, Young S, Weldon RH, Baumgartner A, Anderson D, Wyrobek AJ. 2012. Micronutrients intake is associated with improved sperm DNA quality in older men. Fertility and Sterility 98(5):1130-1137. e1.
Singer K, Maley N, Mergian T, DelProposto J, Cho KW, Zamarron BF, Martinez-Santibanez G, Geletka L, Muir L, Wachowiak P. 2015. Differences in Hematopoietic Stem Cells Contribute to Sexually Dimorphic Inflammatory Responses to High Fat Diet Induced Obesity. Journal of Biological Chemistry 290:13250-13262.
Sloley BD, Urichuk LJ, Tywin C, Coutts RT, Pang PK, Shan JJ. 2001. Comparison of chemical components and antioxidant capacity of different Echinacea species. Journal of Pharmacy and Pharmacology 53(6):849-857.
Sozio MS, Lu C, Zeng Y, Liangpunsakul S, Crabb DW. 2011. Activated AMPK inhibits PPAR-α and PPAR-γ transcriptional activity in hepatoma cells. American Journal of Physiology-Gastrointestinal and Liver Physiology 301(4):G739-G747.
Stimpel M, Proksch A, Wagner H, Lohmann-Matthes M. 1984. Macrophage activation and induction of macrophage cytotoxicity by purified polysaccharide fractions from the plant Echinacea purpurea. Infection and Immunity 46(3):845-849.
Szaleczky E, Prechl J, Feh#westeur042#r J, Somogyi A. 1999. Alterations in enzymatic antioxidant defence in diabetes mellitus− a rational approach. Postgraduate medical journal 75(879):13-17.
Tanrikut C, Goldstein M, Rosoff JS, Lee RK, Nelson CJ, Mulhall JP. 2011. Varicocele as a risk factor for androgen deficiency and effect of repair. BJU International 108(9):1480-1484.
Thygesen L, Thulin J, Mortensen A, Skibsted LH, Molgaard P. 2007. Antioxidant activity of cichoric acid and alkamides from Echinacea purpurea, alone and in combination. Food Chemistry 101(1):74-81.
Turchi P. 2015. Prevalence, Definition, and Classification of Infertility. Clinical Management of Male Infertility: 5-11.
Vimalanathan S, Kang L, Amiguet VT, Livesey J, Arnason JT, Hudson J. 2005. Echinacea purpurea. aerial parts contain multiple antiviral compounds. Pharmaceutical Biology 43(9):740-745.
Vincent HK, Taylor AG. 2006. Biomarkers and potential mechanisms of obesity-induced oxidant stress in humans. International Journal of Obesity 30(3):400-418.
WHO, 2000. Obesity and overweight. World Health Organization. Geneva.
Wright C, Milne S, Leeson H. 2014. Sperm DNA damage caused by oxidative stress: modifiable clinical, lifestyle and nutritional factors in male infertility. Reproductive Biomedicine Online 28(6):684-703.
Xiao H, Xie G, Wang J, Hou X, Wang X, Wu W, Liu X. 2013. Chicoric acid prevents obesity by attenuating hepatic steatosis, inflammation and oxidative stress in high-fat diet-fed mice. Food Research International 54(1):345-353.
Yokoi K, Uthus EO, Nielsen FH. 2003. Nickel deficiency diminishes sperm quantity and movement in rats. Biological Trace Element Research 93(1-3):141-153.
Younglai E, Holt D, Brown P, Jurisicova A, Casper R. 2001. Sperm swim-up techniques and DNA fragmentation. Human Reproduction 16(9):1950-1953.
Zoutewelle G, van Wijk R. 1990. Effects of Echinacea purpurea extracts on fibroblast populated collagen lattice contraction. Phytotherapy Research 4(2):77-81.