臺灣博碩士論文加值系統

女性族群到達了一定年紀，會經歷因月經逐漸減少甚至停止進而引發的各種併發症，例如：肥胖、子宮萎縮、體內氧化壓力增加、血液循環變差、體內雌激素及黃體素接受器表現量發生改變等等。隨著全球逐漸高齡化的趨勢，再加上女性族群平均壽命又高於男性族群。未來，更年期婦女的相關問題勢必成為相當重要的課題。目前常用於治療更年期症狀的西藥卻有不少副作用，因此我們想嘗試副作用較為緩和的中草藥。
中藥複合物B401含有六種中藥成分：人參、黃耆、當歸、地黃、旱蓮草及女貞子，本實驗主要是探討中藥複合物B401是否能改善停經症候群的症狀。首先將去卵巢 (ovariectomize) 小鼠分成四組：控制組分別餵食B401和二次水，以及去卵巢組分別餵食B401和二次水。餵食B401的組別均每天以餵食針餵食30毫克/每公斤體重的B401，餵食持續十天，之後檢測小鼠的子宮重量、體重、體表血流量、活性氧類 (reactive oxygen species, ROS)、子宮中超氧歧化酶 (superoxide dismutase 2, SOD2)、雌激素接受器α / β (estrogen receptor, ER α / β) 與黃體素接受器 (progesterone receptor, PR) 的表現量。研究結果顯示，餵食中藥複合物B401不僅減緩去卵巢導致的小鼠子宮重量、血流、SOD2、ERβ以及PR的減少，也可顯著降低去卵巢鼠體內的自由基、子宮中ERα表現量。結果證實，中藥複合物B401可以作為改善更年期婦女之不適症狀的健康補給品。

Women will go through some complications, such as obesity, uterine shrinking, increase in oxidative stress, decrease in blood flow, change in the expression of estrogen and progesterone receptor, etc, for peri- and postmenopausal women when they come to the certain age.
Along with the aging tendency in the world, and female average life expectancy is higher than male, climacteric syndromes is going to be an important issue in the future, but the Modern Medicine which was usually used to treat them got lots of side effects. We, therefore, intend to try more moderate traditional Chinese medicine.
To evaluate the estrogenic effect of the Chinese herbal formula B401, we examined uterine weight, body weight, blood flow, ROS (reactive oxygen species), SOD2 (superoxide dismutase 2), estrogen receptor (ER), progesterone receptor (PR) activation by B401 treatment in vivo ovariectomized mice. The female mice were randomized into four groups: Sham operated group (Sham), saline treated ovariectomized group (OVT) and B401 treated ovariectomized group (B401+OVT). The B401+OVT mice were oral treated with B401 at a dose of 30 mg/kg bw/day for 10 days. Our results showed that oral treatment with the Chinese herbal formula B401 significantly reversed ovariectomy-induced uterine weight reduction. Moreover, the Chinese herbal formula B401 oral treatment significantly increased blood flow, expression of ERβ and PR in the uterus, and decrease in ovariectomized mice ROS, expression of ERα and SOD2 in the uterus. Taken our data together, we suggested that B401 oral treatment could be protective and beneficial for the management of menopausal symptoms.

目錄 1
圖表目錄 3
致謝 4
縮寫表 5
中文摘要 6
ABSTRACT 7
第一章 緒言 9
1. 停經症候群 10
2. 超氧歧化酶2 10
3. 雌激素接受器與黃體素接受器 11
4. B401中藥配方 13
第二章 研究目的 15
第三章 材料方法 17
1. 實驗動物準備 18
2. 中藥複合物B401 19
3. 血流量分析 19
4. 血液中活性氧類分析 19
5. 血液生化值分析 20
6. 免疫組織化學染色 20
7. 西方墨點法 21
8. 統計分析 24
第四章 實驗結果 25
1. B401中藥對去卵巢小鼠子宮重量及體重的影響 26
2. B401中藥對去卵巢小鼠血流情形的影響 27
3. B401中藥對去卵巢小鼠體內氧化自由基的影響 27
4. B401中藥對去卵巢小鼠子宮中超氧歧化酶2表現量的影響 28
5. B401中藥對去卵巢小鼠子宮中雌激素及黃體素接受器表現量
的影響 29
第五章 討論 32
第六章 結論 37
參考文獻 39
實驗圖表 46

1. Palacios S, Henderson VW, Siseles N, et al. Age of menopause and impact of climacteric symptoms by geographical region. Climacteric 2010, 13(5): 419-28.

2. Kim DI, Choi MS, Pak SC, et al. The effects of Sutaehwan-Gami on menopausal symptoms induced by ovariectomy in rats. BMC Complementary and Alternative Medicine 2012, 12: 227-234.

3. Lee H, Choi J, Shin S, et al. Effects of Korean red ginseng (Panaxginseng) on obesity and adipose inflammation in ovariectomized mice. Journal of Ethnopharmacology 2016, 178: 229-237.

4. Roepke TA, Ronnekleiv OK, Kelly MJ. Physiological consequences of membrane-initiated estrogen signaling in the brain. Front Biosci 2011, 16: 1560–1573.

5. Tawfik SH, Mahmoud BF, Saad MI, et al. Similar and additive effects of ovariectomy and diabetes on insulin resistance and lipid metabolism. Biochemistry Research International 2015, 2015 (567945): 1-8.

6. Coelho M, Oliveira T, Fernandes R. Biochemistry of adipose tissue: an endocrine organ. Archives of Medical Science 2013, 9(2): 191-200.

7. Lihn AS, Pedersen SB, Richelsen B. Adiponectin: action, regulation and association to insulin sensitivity. Obesity Reviews 2005, 6(1):13-21.

8. Nigro E, Scudiero O, Monaco ML, et al. New insight into adiponectin role in obesity and obesity-related diseases. BioMed Research International 2014, 2014: 658913.

9. Chandran M, Phillips SA, Ciaraldi T, et al. Adiponectin: more than just another fat cell hormone? Diabetes Care 2003, 26(8): 2442-2450.

10. Pan H, Guo J, Su Z. Advances in understanding the interrelations between leptin resistance and obesity. Physiology & Behavior 2014, 130: 157-169.

11. Machi JF, Dias DS, Freitas SC, et al. Impact of aging on cardiac function in a female rat model of menopause: role of autonomic control, inflammation, and oxidative stress. Clinical Interventions in Aging 2016, 11: 341–350.

12. Tureka FW, Gillette MU. Melatonin, sleep, and circadian rhythms: rationale for development of specific melatonin agonists. Sleep Medicine 2004, 5(6): 523-532.

13. Cui J, Shen Y, Li R. Estrogen synthesis and signaling pathways during aging: from periphery to brain. Trends in Molecular Medicine 2013, 19: 197-209.

14. Yang S, 2008. Molecular cloning and evolution of cuttlefish (Sepioteuthis lessoniana) estrogen receptor. Master’s thesis.

15. Martinkovich S, Shah D, Planey SL, et al. Selective estrogen receptor modulators: tissue specificity and clinical utility. Clinical Interventions in Aging 2014, 9: 1437-1452.

16. Panno ML, Giordano F. Effects of psoralens as anti-tumoral agents in breast cancer cells. World Journal of Clinical Oncology 2014, 5 (3): 348-358.

17. Marco PD, Cirillo F, Vivacqua A, et al. Novel aspects concerning the functional cross-talk between the insulin/IGF-I system and estrogen signaling in cancer cells. Cancer Endocrinology 2015, 6(30): 1-8.

18. Franck MJ. Estrogen and androgen receptors: regulators of fuel homeostasis and emerging targets for diabetes and obesity. Trends in Endocrinology and Metabolism 2011, 22: 24-33.

19. Nilsson S, Mäkelä S, Treuter E, et al. Mechanisms of estrogen action. American Physiological Society 2001, 81: 1535-1565.

20. Wang SE, Lin CL, Hsu CH, et al. Treatment with a herbal formula B401 enhances neuroprotection and angiogenesis in the R6/2 mouse model of Huntington’s disease. Drug Des Devel Ther 2015, 9: 887–900.

21. Cohen I, Beyth Y, Altaras MM, et al. Estrogen and progesterone receptor expression in postmenopausal tamoxifen-exposed endometrial pathologies. Gynecologic Oncology 1997, 67: 8-15.

22. Tavares CB, Gomes-Braga FDCSA, Costa-Silva DR, et al. Expression of estrogen and progesterone receptors in astrocytomas: a literature review. Clinics 2016, 71(8): 481-486.

23. Tulay OK. Estrogen Deficiency and Osteoporosis. Intech, Advances in Osteoporosis 2015, CC BY 3.0 license, Edited Volume, chapter 2.

24. Ahmeda HM, El-Shereefa HK, El-Gendia SS, et al. Leptin, osteocalcin, and bone mineral density in post-hepatitic liver cirrhosis. Arab Journal of Gastroenterology 2010, 11(3): 130-135.

25. Jagtap VR, Ganu JV, Nagane NS. BMD and Serum Intact Osteocalcin in Postmenopausal Osteoporosis Women. Indian J Clin Biochem 2011, 26(1): 70-73.

26. Prabha YS, Ashalata K, Babu PVSSV, et al. A Study of Bone Markers (Serum Calcium, Serum Phosphorus And Serum Alkaline Phosphatase) In Post Menopausal Women In East Godavari District, Andhra Pradesh, India. IOSR Journal of Dental and Medical Sciences 2015, 14(6): 1-3.

27. Weil VP, Cirigliano MD, Battistini M. Herbal Treatments for
Symptoms of Menopause. Turner White Communications 2000, 35-44.

28. Dana SW, Nguyen BT, Wuttke W. Long-Term Effects of Ovariectomy on Osteoporosis and Obesity in Estrogen-Receptor-β-Deleted Mice. Comparative Medicine 2012, 62: 8-13.

29. Seibel MJ. Biochemical Markers of Bone Turnover Part I: Biochemistry and Variability. The Clinical Biochemist Reviews 2005, 26(4): 97–122.

30. Hsu CH, 2016: Therapeutic Potential of Traditional Chinese Medicine in Maganism. Doctoral dissertation.

31. Kang J, Lee N, Ahn Y, et al. Study on improving blood flow with korean red ginseng substances using digital infrared thermal imaging and doppler sonography: randomized, double blind, placebo-controlled clinical trial with parallel design. J Tradit Chin Med 2013, 33(1): 39-45.

32. Zhang L, Fujii S, Kosaka H. Effect of oestrogen on reactive oxygen species production in the aortas of ovariectomized Dahl salt-sensitive rats. Journal of hypertension 2017, 25(2):407-14.

33. Mutinati M, Spedicato M, Manca R, et al. Influence of antibiotic therapy on serum levels of reactive oxygen species in ovariectomized bitches. Journal of veterinary pharmacology and therapeutics 2008, 31(1):18-21.

34. Weihua Z, Saji S, Ma¨kinen S, et al. Proceedings of the National Academy of Science of the United States of America 2000, 97(11): 5936-5941.

35. Frasor J, Barnett DH, Danes JM, et al. Response-Specific and Ligand Dose-Dependent Modulation of Estrogen Receptor (ER) α Activity by ERβ in the Uterus. Endocrinology 2003, 144 (7): 3159-3166.

36. Purves D, Augustine GJ, Fitzpatrick D, et al. Neuroscience 2001, 2nd ed. Sinauer Associates, Inc. Ch. 30.

37. Handa RJ, Ogawa S, Wang JM, et al. Roles for Oestrogen Receptor β in Adult Brain Function. Journal of Neuroendocrinology 2012, 24(1): 160-173.

38. Chung WCJ, Pak TR, Suzuki S, et al. Detection and localization of an estrogen receptor beta splice variant protein (ERβ2) in the adult female rat forebrain and midbrain regions. Journal of Comparative Neurology 2007, 505(3): 249-267.

39. Esqueda MED, Craig T, Hinojosa-Laborde C. Effect of ovariectomy on renal estrogen receptor-α and estrogen receptor-β in young salt-sensitive and -resistant rats. Hypertension 2007, 50(4): 768-772.

40. Jayachandran M, Miller VM. Ovariectomy upregulates expression of estrogen receptors, NOS, and HSPs in porcine platelets. American Journal of Physiology Heart and Circulatory Physiology 2002, 283: H220-H226.

41. Lund TD, Rovis T, Chung WC, et al. Novel actions of estrogen receptor-beta on anxiety-related behaviors. Endocrinology 2005, 146(2):797-807.

42. Chen C, Kalu DN. Modulation of intestinal estrogen receptor by ovariectomy, estrogen and growth hormone. The Journal Of Pharmacology And Experimental Therapeutics 1998, 286: 328-333.

43. Sarrel PM. Ovarian hormones and the circulation. Maturitas 1990, 12(3): 287-298.

44. Mercuro G, Longu G, Zoncu S, et al. Impaired forearm blood flow and vasodilator reserve in healthy postmenopausal women. American Heart Journal 1999, 137(4): 692-697.

45. Shuvy M, Abedat S, Beeri R, et al. Raloxifene attenuates Gas6 and apoptosis in experimental aortic valve disease in renal failure. American journal of physiology Heart and circulatiory physiology 2011, 300(5): H1829-1840.

46. Gu X, Masters KS. Role of the MAPK/ERK pathway in valvular interstitial cell calcification. American journal of physiology Heart and circulatiory physiology 2009, 296(6): H1748-1757.

47. Chikahisa Higuchi, Akira Myoui, Nobuyuki Hashimoto, et al. Continuous inhibition of mapk signaling promotes the early osteoblastic differentiation and mineralization of the extracellular matrix. Journal Of Bone And Mineral Research 2002, 17(10): 1785-1794.