前人研究成果提出，一氧化氮合成脢確實存在與骨骼組織中三種最主要的細胞：骨細胞、造骨細胞與破骨細胞，而不論是離體細胞培養實驗或是活體骨骼組織機械應力刺激實驗均發現，骨骼組織細胞在受到應力刺激後會產生多量的一氧化氮，而一氧化氮也被證實與破骨細胞、造骨細胞之間的相互調控有密不可分的關係，而運動訓練過程中對於骨骼組織產生的影響是否就是經由一氧化氮合成脢作為傳導的媒介？而雌激素與運動訓練對保護骨骼組織產生的加成性效果是否也與一氧化氮合成脢有關，此二疑點及為本研究之主要目的，本研究結合運動訓練與雌激素補充治療，並以一氧化氮合成脢抑制劑交互使用，以期探討一氧化氮合成脢在整個訊息傳導過程中扮演的角色與定位。八十隻十二週齡之大白鼠隨機分派成八組，分別給予運動訓練、雌激素補充以及一氧化氮合成脢抑制劑或合併運動訓練與雌激素補充、運動訓練以及一氧化氮合成脢抑制劑、雌激素補充合併一氧化氮合成脢抑制劑之處理，經過十二週原地跑步機之運動訓練後分別進行股骨頸耐壓能力、股骨頸斷裂部位X光吸光值、一氧化氮合成脢表現量之分析。研究結果：（1） 合併運動訓練與雌激素補充治療之大鼠骨骼組織中內皮型一氧化氮合成脢與誘發型一氧化氮合成脢表現量高於單獨使用雌激素治療或運動訓練之大鼠。（2） 使用一氧化氮合成脢抑制劑卻直阻斷運動訓練保護骨質流失的效果，但對雌激素補充治療的阻斷效果較不明顯。本研究提出下列兩點結論：
（1） 一氧化氮合成脢確實在運動訓練影響骨骼組織的訊息傳導過程專扮演不可或缺的關鍵角色。
（2） 雌激素補充治療與運動訓練合併使用產生的加成性效應可能尚有其他代償機轉，並不一定完全經由一氧化氮合成脢轉介。

The modulator between osteoblast and osteoclast used to be as inflammatory cytokines such as interlukin-6, TNF and IGF. In passed years, spotlight of bone modulator had turned to be nitric oxide, a short-lived free radical that is generated by the NO synthase group of enzymes which syntheses NO by combine molecular oxygen with the terminal guanidino nitrogen of the amino acid L-arginine. Due to the small molecular size, NO can pass through most kind of membrance easily and act on guanidine cyclase. By changing conformation of guanidine cyclase and altering cGMP level of cytosol, NO affects cell proliferation or activity such as osteoclast. Three isoforms of nitric oxide synthase has been identified , however , only two of them were expressed in bone tissue : endothelial form (eNOS) and induce form (iNOS). Recent study had indicated that osteoblast and osteocyte mainly express endothelial form of NOS under normal conditions, while osteoclast express not only eNOS but also iNOS , even though the later is much weakly.
Recent study had confirmed the additive effect of exercise training and ERT. Nitric oxide synthase might play an important role in this process. In this study, we assume that nitric oxide synthase is the key enzyme which mediate signal transduction of estrogen and mechanical strain. Eighty Spraque-Dawley rats were randomly assign to eight groups, which treated with exercise training, Estrogen replacement therapy(ERT), nitric oxide synthase inhibitor(NOS inhibitor), Exercxuse and ERT, Exercise and NOS inhibitor or ERT and NOS inhibitor, respectively. The data of mechanical and biochemical test wee collected and analysed by SPSS9.0, results were followed:
(1) Both ERT and exercise training enhance nitric oxide synthase expression, combine two method shows significant additive effect.
(2) Nitric oxide synthase inhibitor suppresses mechanically induced bone formation and reduces maximal loading force of femur neck

林嘉志。（民88）運動訓練對氧化態低密度脂蛋白抑制流體媒介之血管舒張作用的生理影響。國立台灣師範大學體育研究所。未發表之博士論文。
黃滄海。（民90）不同強度的耐力性游泳運動對大鼠骨骼發展的影響。國立台灣師範大學體育研究所。未發表之博士論文。
張豐麟。（民90）持續性與間歇式動訓練對大鼠骨骼發展的影響。國立台灣師範大學體育研究所。未發表之碩士論文。
Armour E. kathatine and Ralston H. Stuarth (1998) Estrogen upregulate endothelial constitutive nitric oxide synthase express in human osteoblast-like cells. Endocrinology 139(2), 799-783.
Ashish D. Diwan, Min X. Wang, Daniel jang, Wei zhu, and George A.C. murrell (2000) Nitric oxide modulate fracture healing. Journal of bone and mineral research 15(2), 342-351.
Baldock A.J. Paul, Need G.Allan, Moore J.Robert, Durbridge C. Timothy and Morris A. Howard.(1999) Discordance between bone turnover and bone loss : Effects of aging and ovarietomy in the rat. Journal of Bone and Mineral Research 14(8), 1442-1448.
Barry S. Komm, Chrsitopher M. Terpening, David J. Benz, Kimberlie A. Graeme, Alfred Gallegos, Murray Korc, Geoffrey L. Greene, Bert W. O‘Malley, Mark R. Haussler (1988) Estrogen Binding, Receptor mRNA, and Biologic Response in Osteoblast-Like Osteosarcoma Cells. Science, 24, 81-83.
Barengolts EI. Curry DJ. Bapna MS. Kukreja SC.(1993) Effects of two non-endurance exercise protocols on established bone loss. Calcified Tissue International, 52(3), 239-43.
Barengolts EI. Lathon PV. Curry DJ. Kukreja SC.(1994) Effects of endurance exercise on bone histomorphometic parameters in intact and ovariectomized rats. Bone and Mineral, 26(2), 133-40.
Barenglots EI. Curry DJ. Bapna MS. Kukreja Sc.(1993) Effects of endurance exercise on bone mass and mechanical properties in intect and ovariectomized rats. Journal of Bone and Mineral Research, 8(8), 937-42.
Donahue, R.S. Mazzeo, and S.M. Horvath (1988) Endurance Training and Bone Loss in Calcium-Deficient and Ovariectomized Rats. Metabolism, 37, 741-744
Diane M. Raab, Evertt L. Smith, Thomas D. Crenshaw, and D.Paul Thomas (1990) Bone mechanical properties after exercise training in young and old rats. American Journal of Physiological,130-133
Dike N. Kalu (1991) The ovariectomized rat model of postmenopausal bone loss. Bone and Mineral, 15 175-192
Dalia Somjen,Itzhak Binderman, Esther Berger and Arie Harell (1979) Bone Remodelling Induced by Physical Stress is Prostaglandin E2 Mediated. 91-99
Evans M. Deborah and Ralston H. Stuart.(1996) Nitric and bone. Journal of Bone and Mineral Research, 11(3), 300-305
Everett L. Smith and Lorri Tommerup (1995) Exercise: A Prevention and Treatment for Osteoposis and Injurious Falls in the Older Adult. Journal of Aging and Physical Activity, 3, 178-192
Erman Adlercreutz, Hideo Honjo, Akane Higashi, Theodore Fotsis, Esa Hamalainen, Takeshi Hasegawa, and Hiroji Okada (1991) Urinary Excretion of lignans and isoflavonoid phytoestrogens in Janpanese men and women consuming a traditional Janpanese diet .Am Journal Clin Nur, 54, 1093-1100
Erik F. Eriksen, Dougias S. Colvard, Nicholas J. Berg, Mark L. Graham, Kenneth G. Mann, Thomas C. Spelsberg, B. Lawrence Riggs (1988) Evidence of Estrogen Receptors in Normal Human Osteoblast-Like Cells. Science, 24,84-86
Fuller,A.C.Gallagher and T.J.Chambers (1991) Osteoclast Resorption - Stimulating Activity is Associated With the Osteoblast Cell Surface and/or the Extracellular Matrix. Biochemical and Biophysical Research Communications, 181, 67-73
Giuseppe Girasole, Robert L.Jika, Giovanni Passeri, Scott Boswell, George Boder, Daniel C.Williams, and Stavros C. Manolagas (1992) 17β-Estradiol Inhibits Interleukin-6 Production by Bone Marrow-derived Stromal Cells and Osteoblasts In Vitro: A Potential Mechanism for the Antiosteoporotic Effect of Estrogens. The Journal of Clinical Investigation, 89, 883-891
Harold M. Frost (1999) On the estrogen –bone relationship and postmenopausal bone loss : A new model. Journal of Bone and Mineral Research, 14(9), 1473-1477
Helfrich H. Miep, Evans E.Deborah, Grabowski S. Peter, Pollock S. Jennifer, Ohshima Hiroshi, and Ralston Stuart. (1997) Express of nitric oxide synthase isoforms in bone and bone cell cultures. Journal of Bone and Mineral Research 12(7), 1108-1115.
Hirokazu Tsukahara, Masakazu Miura, Shinya Tsuchida, Ikue Hata, Keishi Hata, Kazutaka Yamamoto, Yasushi Ishit, Ikunobu Muramatsu, and Masakatsu Sudo. (1996) Effect of Nitric Oxide Synthase Inhabitors on Bone Metabolism in Growing Rats. Endocrinology Metablism, 33, E840-E845
Holliday L. Shannon, Dean D. Alan, Lin H. Royana, Greenwald E. James, and Gluck L. Stephen. (1997) Low NO concentrarion inhibit osteoclast formation in mouse marrow cultures by cGMP-dependent mechanism. American Journal of Physiology 272(renal physiology 41), F283-291.
Holly X. and Zerath E. (2000) Bone mass increase in less than 4 weeks of voluntary exercising in growing rats. Medicine and Science in Sports and Exercise, 32(9), 1562-1569
Hukkanen V.J. Mika, Platts A.M.Louise, Inigi Fernandez DE Marticorena, O'Shaughnessy Meg, Macityre Iain, and Polak M. Julia.(1999) Developmental regulation of nitric oxide synthase expression in rat skeletal bone. Journal of Bone and Mineral Research 14(6), 868-877.
Jean Yves Reginster, Nathalie Sarlet, Rita Deroisy, Adelin Albert, Uysse Gaspard, and Paul Franchimont (1992) Minimal Levels of Serum Estradiol Prevent Postmenopausal Bone Loss. Calcified Tissue Internatioanal, 51, 340-343
J. Tuukkanen, Zhiqi Peng, H.K. Vaananen (1994) Effect of Running Exercise on the Bone Loss Induced by Orchidectomy in the Rat. Calcified Tissue International, 55, 33-37
James K.Yeh, Chung C. Liu, and John F. Aloia (1993) Additive Effect of Treadmill Exercise and 17β-Estradiol Replacement on Prevention of Tibial Bone Loss in Adult Ovariectomized rat. Journal of Bone and Mineral Research, 8, 677-682
Jack C.-H. Hou,M.D.,Ph D, Ronald F.Zernicke, Ph.D.,and R.James Barnard, Ph.D.(1989) Experiment Diabetes,Insulin Treatment, and Femoral Neck Morphology and Biomechanics in Rats.
Jack C.-H. Hou, George J.Salem,Ronald F. Zernicke, and R James Barnard (1990) Structural and mechanical adaptations of immature trabecular bone to strenuous exercise. American Journal of Physiology,1309-1313
Janet M. Hock,Michael Centrella, and Ernesto Canalis (1988) Insulin-Like Growth Factor I Has Independent Effects on Bone Matrix Formation and Cell Replication. Endocrinology, 254-259
Kohrt W.M., Snead D.B., Slatopolsky E., Birge S.J. Jr. (1995) Addictvie effects of weight-bearing exercise and estrogen on bone mineral density in older women. Journal of Bone and Mineral Research, 10(9), 1303-1311
Kohtaro kawashima, Takeshi Inoue,Naoyuki Tsutsumit and Hiroyoshi Endo (1996) Effect of KCA-098 on the Function of Osteoblast-Like Cells and the Foramtion of TRAP-Positive Multinucleated Cells in a Mouse Bone Marrow Cell Population. Biochemical Pharmacology, 51, 133-139
Kathleen M. Reich and John A. Frangos (1991) Effect of flow on prostaglandin E2 and inositoltrisphosphate levels in osteoblasts. American Journal of Physiology,428-431
Lane E. Nancy, David Haupt, Donald B. Kimmel. Gunnar Modin, and John H. Kinney. (1999) Early estrogen replacement therapy reverse the rapid loss of trabecular bone volume and prevents further deterioration of connectivity in the rats. Journal of Bone and Mineral Research 14(2), 206-214.
Lipton S.A., Choi Y., Pan Z.(1993) A redox-based mechanism for the neuroprotective and neudestructive effects of nitric oxide and related nitroso-compounds. Nature 364:626-632
Löwik CWGM, Nibbering P.H., Van der Ruit M., Papapoulos S.E.(1994) Inducible production of nitric oxide in osteoblast-like cells and in fetal bone explants is associated with suppression of osteoclast bone resorpsion. Journal of clinical Investigation, 93,1465-1472
Mcallister T.N. and Frangos J.A. (1999) Steady and transient fluid shear stress stimulate NO release in osteoblast through distinct biochemical pathway. Journal of Bone and Mineral Research 14(6), 930-936
Mancini L., Bidhendi Moradi, Brandi M.L., and MacIntyre I.(1998) Nirtic oxide superoxide and peroxynitrite modulate osteoclast activity. Biochemical and Biophysical Research Communications, 243(2), 785-790
Momcada S., Higgs A. (1993) The L-arginine-nitric oxide pathway. New England Journal of Medicine. 329, 2002-2012
Mori S, Jee WSS, Li XJ, Chan S, Kimmel DB.(1990)Effect of Prostaglandin E2 on production of new cancellous bone in the axial skeleton of ovariectomizedrat. BONE 11, 103-113.
Masami Kojima, Naoyuki Tsutsumi, Hideo Nagata,Fumiaki Itoh, Arao Ujiie, Kohtaro kawashima, Hiroyoshi Endo, and Mitsuo Okazaki (1994) Effect of KCA-098, a New Benzofuriquinoline Derivative, on Bone Mineral Metabolism. Biological Pharmacological Bulletin, 17, 504-508
Naoyuki Takahashi,Takuhiko Akatsu,Nobuyuki Udagawa, Takashisa Sasaki, Akria Yamaguchi, Jane M. Moseley, T. John Martin and Tatsuo Suda (1988) Osteoblastic Cells Are Involved in Osteolast Formation. Endocrinology, 2600-2602
Naoyuki Tsutsumi(1995) Effect of Coumestrol on Bone Metabolism in Organ Culture. Biological Pharmacological Bulletin, 18, 1012-1015
Om P. Sharma, Herman Adlercreutz, John D. Strandberg, Barry R. Zirkin,DOnald S. Coffey and Larry L.Ewing (1992) Soy of Dietary Source Plays a Role Against The Pathogenesis of Prostatitis in Rats. J. Steroid Biochem. Biol, 43, 557-564
Pai Chin Kao, Fang-Ku Peng (1995) How to Reduce the Rick Factors of Osteoporosis in Asia. Chin Med Journal (Taipei), 55, 209-13
Philip D. Chilibeck, Digby G. Sale and Colin E. Webber (1995) Exercise ang Bone Mineral Density. Sports Medicine, 19(2), 103-122
Pohlman, L.A. Darby, and A.J. Lechner (1985) Morphometry and calcium contents in appendicular and axial bones of exercised ovariectomized rats. American Journal of Physiology,12-17
Reid, M.B. (1998) Role of nitric oxide in skeleton muscle: sybthesis, distribution amd functional importance. Acta Physiologica Scandinavica, 162, 401-409
Robert L.Jika,Giao Hangoc,Giuseppe Girasole, Giovanni Passeri, Daniel C.Williams, John S. Abrams, Brendan Boyce, Hal Broxmeyer,Stavros C.Manolagas (1992) Increased Osteoclast Development After Estrogen Loss: Mediation by Interleukin-6. Science, 257 88-91
Szucs (1994) Osteoporosis from the point of view of the clinical endocrinologist. Agents Actions, 41 80-83
Tomoe Ishii, Tomoyuki Saito, Kyoko Morimoto, Yasuhiro Takeuchi, Shigetaka Asano, Masayoshi Kumegawa,Etsuro Ogata and Toshio Matsumoto (1993) Estrogen Stimulates The Elaboration of Cell/Matrix Surface-Associated Inhibitory Factor of Osteoclastic Bone Resorption From Osteoblastic Cells. Biochemical and Biophysical Research Communications, 191, 495-502
William R. Phipps,Margaret C. Martini, Johnna W. Lampe, Joanne L. Slavin, and Mindy S. Kurzer (1993) Effect of flax Seed Ingestion on the Menstrual Cycle. Journal of Clinical Endocrinology and Metabolism, 77, 1215-1219
Wimalawansa S.J., DeMarco G., Gangula P., and Yallampalli C. (1996) Nitric oxide donor alleviates ovariectomy-induced bone loss. Bone18(4), 301-304
Yang S. Chyun and Lawrence G.Raisz (1984) Stimulation if Bone Formation by Prostaglandin E2. American Journal of Physiology, 27, 97-101
Yeh J.K., Aloia J.F., Barilla ML. (1994) Effects of beta-estradiol replacement and treadmill exercise on vertebral and femoral bones of the ovariectomized rat. Bone and Mineral, 24(3), 223-34n
Yoshiko Ishimi,Chisato Miyaura,Cheng He Jin, Takuhiko Akatsu,Etsuko Abe,Yuchi Nakamura,Akira Yamaguchi,Shusaku Yoshiku, Tadashi Matsuda,Toshio Hirano, Tadamtsu Kishimoto,and Tatsuo Suda (1990) IL-6 Produced by Osteoblasts and Induces Bone Resorption. The Journal of Immunology, 145, 3297-3303
Zaman G., Pitsillides A.A., Rawlinson S.C.F., Suswillo R.F.L., Mosley J.R., Cheng M.Z., Platts L.A.M., Hukkanen M., Polak J.M., and Lanyon L.E. (1999) Mechanical strain stimulates nitric oxide production by rapid activation of endothelial nitric oxide synthase in osteocyte. Journal of Bone and Mineral Research 14(7), 1123-1131
Zamne G., Cheng M.Z., Jessop H.L., White R., and Lanyon L.E. (2000) Mechanical strain activates estrogen response elements in bone cells. Bone, 27(2), 233-239
Zhiqi Peng, Juha Tuukkanen, and H. Kalervo Vaananen (1994) Exercise Can Provide Protection Against Bone Loss and Prevent the Decrease in Mechanisal Strength of Fremoral Neck in Ovariectomized Rats. Journal of Bone and Mineral Research, 9, 1559-1564