Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/23172
標題: 金線連糖苷對小鼠內毒素休克、風濕性關節炎和骨質疏鬆之改善作用
Ameliorative effects of kinsenoside on endotoxic shock, rheumatoid arthritis and osteoporosis in mice
作者: 蕭宏柏
Hsiao, Hung-Bo
關鍵字: kinsenoside;金線連糖苷抗發炎;inflammatory
出版社: 生命科學系所
引用: Angus, D. C., W. T. Linde-Zwirble, et al. (2001). "Epidemiology of severe sepsis in the United States: analysis of incidence, outcome, and associated costs of care." Crit Care Med 29(7): 1303-10. Asagiri, M., K. Sato, et al. (2005). "Autoamplification of NFATc1 expression determines its essential role in bone homeostasis." J Exp Med 202(9): 1261-9. Asagiri, M. and H. Takayanagi (2007). "The molecular understanding of osteoclast differentiation." Bone 40(2): 251-64. Badolato, R. and J. J. Oppenheim (1996). "Role of cytokines, acute-phase proteins, and chemokines in the progression of rheumatoid arthritis." Semin Arthritis Rheum 26(2): 526-38. Baldwin, A. S., Jr. (1996). "The NF-kappa B and I kappa B proteins: new discoveries and insights." Annu Rev Immunol 14: 649-83. Belkaid, Y., C. A. Piccirillo, et al. (2002). "CD4+CD25+ regulatory T cells control Leishmania major persistence and immunity." Nature 420(6915): 502-7. Berlato, C., M. A. Cassatella, et al. (2002). "Involvement of suppressor of cytokine signaling-3 as a mediator of the inhibitory effects of IL-10 on lipopolysaccharide-induced macrophage activation." J Immunol 168(12): 6404-11. Biskobing, D. M., X. Fan, et al. (1995). "Characterization of MCSF-induced proliferation and subsequent osteoclast formation in murine marrow culture." J Bone Miner Res 10(7): 1025-32. Blei, F., E. L. Wilson, et al. (1993). "Mechanism of action of angiostatic steroids: suppression of plasminogen activator activity via stimulation of plasminogen activator inhibitor synthesis." J Cell Physiol 155(3): 568-78. Boonstra, A., R. Rajsbaum, et al. (2006). "Macrophages and myeloid dendritic cells, but not plasmacytoid dendritic cells, produce IL-10 in response to MyD88- and TRIF-dependent TLR signals, and TLR-independent signals." J Immunol 177(11): 7551-8. Borovikova, L. V., S. Ivanova, et al. (2000). "Vagus nerve stimulation attenuates the systemic inflammatory response to endotoxin." Nature 405(6785): 458-62. Brand, D. D., A. H. Kang, et al. (2003). "Immunopathogenesis of collagen arthritis." Springer Semin Immunopathol 25(1): 3-18. Calandra, T., B. Echtenacher, et al. (2000). "Protection from septic shock by neutralization of macrophage migration inhibitory factor." Nat Med 6(2): 164-70. Castellheim, A., O. L. Brekke, et al. (2009). "Innate immune responses to danger signals in systemic inflammatory response syndrome and sepsis." Scand J Immunol 69(6): 479-91. Cavaillon, J. M. and M. Adib-Conquy (2005). "Monocytes/macrophages and sepsis." Crit Care Med 33(12 Suppl): S506-9. Cawston, T. (1998). "Matrix metalloproteinases and TIMPs: properties and implications for the rheumatic diseases." Mol Med Today 4(3): 130-7. Chu, C. Q. and M. Londei (1999). "Differential activities of immunogenic collagen type II peptides in the induction of nasal tolerance to collagen-induced arthritis." J Autoimmun 12(1): 35-42. Coussens, L. M., C. L. Tinkle, et al. (2000). "MMP-9 supplied by bone marrow-derived cells contributes to skin carcinogenesis." Cell 103(3): 481-90. Cross, A. S., S. M. Opal, et al. (1993). "Choice of bacteria in animal models of sepsis." Infect Immun 61(7): 2741-7. Crowston, J. G., M. Salmon, et al. (1997). "T-lymphocyte-fibroblast interactions." Biochem Soc Trans 25(2): 529-31. Dean, D. D., J. Martel-Pelletier, et al. (1989). "Evidence for metalloproteinase and metalloproteinase inhibitor imbalance in human osteoarthritic cartilage." J Clin Invest 84(2): 678-85. Delaisse, J. M., M. T. Engsig, et al. (2000). "Proteinases in bone resorption: obvious and less obvious roles." Clin Chim Acta 291(2): 223-34. Di Giovine, F. S., G. Nuki, et al. (1988). "Tumour necrosis factor in synovial exudates." Ann Rheum Dis 47(9): 768-72. Dolhain, R. J., A. N. van der Heiden, et al. (1996). "Shift toward T lymphocytes with a T helper 1 cytokine-secretion profile in the joints of patients with rheumatoid arthritis." Arthritis Rheum 39(12): 1961-9. Dong, Q., R. Fan, et al. (2009). "Over-expression of SOCS-3 gene promotes IL-10 production by JEG-3 trophoblast cells." Placenta 30(1): 11-4. Engsig, M. T., Q. J. Chen, et al. (2000). "Matrix metalloproteinase 9 and vascular endothelial growth factor are essential for osteoclast recruitment into developing long bones." J Cell Biol 151(4): 879-89. Fink, M. P. and S. O. Heard (1990). "Laboratory models of sepsis and septic shock." J Surg Res 49(2): 186-96. Franzoso, G., L. Carlson, et al. (1997). "Requirement for NF-kappaB in osteoclast and B-cell development." Genes Dev 11(24): 3482-96. Fujiwara, N. and K. Kobayashi (2005). "Macrophages in inflammation." Curr Drug Targets Inflamm Allergy 4(3): 281-6. Galis, Z. S. and J. J. Khatri (2002). "Matrix metalloproteinases in vascular remodeling and atherogenesis: the good, the bad, and the ugly." Circ Res 90(3): 251-62. Galis, Z. S., G. K. Sukhova, et al. (1994). "Increased expression of matrix metalloproteinases and matrix degrading activity in vulnerable regions of human atherosclerotic plaques." J Clin Invest 94(6): 2493-503. Gangi, E., C. Vasu, et al. (2005). "IL-10-producing CD4+CD25+ regulatory T cells play a critical role in granulocyte-macrophage colony-stimulating factor-induced suppression of experimental autoimmune thyroiditis." J Immunol 174(11): 7006-13. Garrington, T. P. and G. L. Johnson (1999). "Organization and regulation of mitogen-activated protein kinase signaling pathways." Curr Opin Cell Biol 11(2): 211-8. Gordon, S. (2003). "Alternative activation of macrophages." Nat Rev Immunol 3(1): 23-35. Gordon, S. (2007). "The macrophage: past, present and future." Eur J Immunol 37 Suppl 1: S9-17. Graesser, D., S. Mahooti, et al. (2000). "Distinct roles for matrix metalloproteinase-2 and alpha4 integrin in autoimmune T cell extravasation and residency in brain parenchyma during experimental autoimmune encephalomyelitis." J Neuroimmunol 109(2): 121-31. Graves, D. T., C. P. Chen, et al. (2000). "Interleukin-1 receptor signaling rather than that of tumor necrosis factor is critical in protecting the host from the severe consequences of a polymicrobe anaerobic infection." Infect Immun 68(8): 4746-51. Ha, H., H. B. Kwak, et al. (2004). "Reactive oxygen species mediate RANK signaling in osteoclasts." Exp Cell Res 301(2): 119-27. Hadler-Olsen, E., B. Fadnes, et al. (2011). "Regulation of matrix metalloproteinase activity in health and disease." FEBS J 278(1): 28-45. Halleen, J. M., S. Raisanen, et al. (1999). "Intracellular fragmentation of bone resorption products by reactive oxygen species generated by osteoclastic tartrate-resistant acid phosphatase." J Biol Chem 274(33): 22907-10. Harris, M. L., H. J. Schiller, et al. (1992). "Free radicals and other reactive oxygen metabolites in inflammatory bowel disease: cause, consequence or epiphenomenon?" Pharmacol Ther 53(3): 375-408. Holmes, C. L., J. A. Russell, et al. (2003). "Genetic polymorphisms in sepsis and septic shock: role in prognosis and potential for therapy." Chest 124(3): 1103-15. Hotokezaka, H., E. Sakai, et al. (2002). "U0126 and PD98059, specific inhibitors of MEK, accelerate differentiation of RAW264.7 cells into osteoclast-like cells." J Biol Chem 277(49): 47366-72. Hsieh, C. C., H. B. Hsiao, et al. (2010). "A standardized aqueous extract of Anoectochilus formosanus modulated airway hyperresponsiveness in an OVA-inhaled murine model." Phytomedicine 17(8-9): 557-62. Ip, Y. T. and R. J. Davis (1998). "Signal transduction by the c-Jun N-terminal kinase (JNK)--from inflammation to development." Curr Opin Cell Biol 10(2): 205-19. Jones, C. B., D. C. Sane, et al. (2003). "Matrix metalloproteinases: a review of their structure and role in acute coronary syndrome." Cardiovasc Res 59(4): 812-23. Jovanovic, D. V., J. A. Di Battista, et al. (1998). "IL-17 stimulates the production and expression of proinflammatory cytokines, IL-beta and TNF-alpha, by human macrophages." J Immunol 160(7): 3513-21. Ju, J. H., M. L. Cho, et al. (2008). "Oral administration of type-II collagen suppresses IL-17-associated RANKL expression of CD4+ T cells in collagen-induced arthritis." Immunol Lett 117(1): 16-25. Kelchtermans, H., L. Geboes, et al. (2009). "Activated CD4+CD25+ regulatory T cells inhibit osteoclastogenesis and collagen-induced arthritis." Ann Rheum Dis 68(5): 744-50. Kim, K., S. H. Lee, et al. (2008). "NFATc1 induces osteoclast fusion via up-regulation of Atp6v0d2 and the dendritic cell-specific transmembrane protein (DC-STAMP)." Mol Endocrinol 22(1): 176-85. Kim, S. H., E. R. Lechman, et al. (2005). "Exosomes derived from IL-10-treated dendritic cells can suppress inflammation and collagen-induced arthritis." J Immunol 174(10): 6440-8. Kirkham, B. W., M. N. Lassere, et al. (2006). "Synovial membrane cytokine expression is predictive of joint damage progression in rheumatoid arthritis: a two-year prospective study (the DAMAGE study cohort)." Arthritis Rheum 54(4): 1122-31. Kyriakis, J. M. and J. Avruch (1996). "Sounding the alarm: protein kinase cascades activated by stress and inflammation." J Biol Chem 271(40): 24313-6. Li, Q. and I. M. Verma (2002). "NF-kappaB regulation in the immune system." Nat Rev Immunol 2(10): 725-34. Lim, H. W., P. Hillsamer, et al. (2005). "Cutting edge: direct suppression of B cells by CD4+ CD25+ regulatory T cells." J Immunol 175(7): 4180-3. Liu, Y. W., C. C. Chen, et al. (2006). "Lipopolysaccharide-induced transcriptional activation of interleukin-10 is mediated by MAPK- and NF-kappaB-induced CCAAT/enhancer-binding protein delta in mouse macrophages." Cell Signal 18(9): 1492-500. Lomaga, M. A., W. C. Yeh, et al. (1999). "TRAF6 deficiency results in osteopetrosis and defective interleukin-1, CD40, and LPS signaling." Genes Dev 13(8): 1015-24. Lubberts, E., M. I. Koenders, et al. (2004). "Treatment with a neutralizing anti-murine interleukin-17 antibody after the onset of collagen-induced arthritis reduces joint inflammation, cartilage destruction, and bone erosion." Arthritis Rheum 50(2): 650-9. Ma, Y. and R. M. Pope (2005). "The role of macrophages in rheumatoid arthritis." Curr Pharm Des 11(5): 569-80. Maini, R. N., M. J. Elliott, et al. (1995). "Beneficial effects of tumour necrosis factor-alpha (TNF-alpha) blockade in rheumatoid arthritis (RA)." Clin Exp Immunol 101(2): 207-12. Martel-Pelletier, J., D. J. Welsch, et al. (2001). "Metalloproteases and inhibitors in arthritic diseases." Best Pract Res Clin Rheumatol 15(5): 805-29. Massova, I., L. P. Kotra, et al. (1998). "Matrix metalloproteinases: structures, evolution, and diversification." FASEB J 12(12): 1075-95. Masuda, K., M. Ikeuchi, et al. (2008). "Suppressive effects of Anoectochilus formosanus extract on osteoclast formation in vitro and bone resorption in vivo." J Bone Miner Metab 26(2): 123-9. Matusevicius, D., P. Kivisakk, et al. (1999). "Interleukin-17 mRNA expression in blood and CSF mononuclear cells is augmented in multiple sclerosis." Mult Scler 5(2): 101-4. Mayeux, P. R. (1997). "Pathobiology of lipopolysaccharide." J Toxicol Environ Health 51(5): 415-35. Miyazaki, T., H. Katagiri, et al. (2000). "Reciprocal role of ERK and NF-kappaB pathways in survival and activation of osteoclasts." J Cell Biol 148(2): 333-42. Mohamed, S. G., E. Sugiyama, et al. (2007). "Interleukin-10 inhibits RANKL-mediated expression of NFATc1 in part via suppression of c-Fos and c-Jun in RAW264.7 cells and mouse bone marrow cells." Bone 41(4): 592-602. Molet, S., Q. Hamid, et al. (2001). "IL-17 is increased in asthmatic airways and induces human bronchial fibroblasts to produce cytokines." J Allergy Clin Immunol 108(3): 430-8. Morgan, M. E., R. Flierman, et al. (2005). "Effective treatment of collagen-induced arthritis by adoptive transfer of CD25+ regulatory T cells." Arthritis Rheum 52(7): 2212-21. Muriel, P. and Y. Escobar (2003). "Kupffer cells are responsible for liver cirrhosis induced by carbon tetrachloride." J Appl Toxicol 23(2): 103-8. Nelson, A. R., B. Fingleton, et al. (2000). "Matrix metalloproteinases: biologic activity and clinical implications." J Clin Oncol 18(5): 1135-49. O''Garra, A., F. J. Barrat, et al. (2008). "Strategies for use of IL-10 or its antagonists in human disease." Immunol Rev 223: 114-31. O''Toole, T. E., Y. T. Zheng, et al. (2009). "Acrolein activates matrix metalloproteinases by increasing reactive oxygen species in macrophages." Toxicol Appl Pharmacol 236(2): 194-201. Okada, Y., N. Takeuchi, et al. (1989). "Immunolocalization of matrix metalloproteinase 3 (stromelysin) in rheumatoid synovioblasts (B cells): correlation with rheumatoid arthritis." Ann Rheum Dis 48(8): 645-53. Pahl, H. L. (1999). "Activators and target genes of Rel/NF-kappaB transcription factors." Oncogene 18(49): 6853-66. Pope, R. M. and J. Tschopp (2007). "The role of interleukin-1 and the inflammasome in gout: implications for therapy." Arthritis Rheum 56(10): 3183-8. Rajashekhar, G., M. Kamocka, et al. (2010). "Pro-inflammatory angiogenesis is mediated by p38 MAP kinase." J Cell Physiol 226(3): 800-8. Ramnath, R. D., S. W. Ng, et al. (2008). "Role of MCP-1 in endotoxemia and sepsis." Int Immunopharmacol 8(6): 810-8. Rank, N., C. Michel, et al. (2000). "N-acetylcysteine increases liver blood flow and improves liver function in septic shock patients: results of a prospective, randomized, double-blind study." Crit Care Med 28(12): 3799-807. Redlich, K., S. Hayer, et al. (2002). "Osteoclasts are essential for TNF-alpha-mediated joint destruction." J Clin Invest 110(10): 1419-27. Roodman, G. D. (1996). "Advances in bone biology: the osteoclast." Endocr Rev 17(4): 308-32. Sakurai, H., H. Chiba, et al. (1999). "IkappaB kinases phosphorylate NF-kappaB p65 subunit on serine 536 in the transactivation domain." J Biol Chem 274(43): 30353-6. Sen, R. and D. Baltimore (1986). "Inducibility of kappa immunoglobulin enhancer-binding protein Nf-kappa B by a posttranslational mechanism." Cell 47(6): 921-8. Senftleben, U. and M. Karin (2002). "The IKK/NF-kappaB pathway." Crit Care Med 30(1 Supp): S18-S26. Shevach, E. M. (2002). "CD4+ CD25+ suppressor T cells: more questions than answers." Nat Rev Immunol 2(6): 389-400. Shih, C. C., Y. W. Wu, et al. (2004). "Effect of Anoectochilus formosanus on fibrosis and regeneration of the liver in rats." Clin Exp Pharmacol Physiol 31(9): 620-5. Shih, C. C., Y. W. Wu, et al. (2001). "Ameliorative effects of Anoectochilus formosanus extract on osteopenia in ovariectomized rats." J Ethnopharmacol 77(2-3): 233-8. Song, I., J. H. Kim, et al. (2009). "Regulatory mechanism of NFATc1 in RANKL-induced osteoclast activation." FEBS Lett 583(14): 2435-40. Steiner, G., M. Tohidast-Akrad, et al. (1999). "Cytokine production by synovial T cells in rheumatoid arthritis." Rheumatology (Oxford) 38(3): 202-13. Sternlicht, M. D., M. J. Bissell, et al. (2000). "The matrix metalloproteinase stromelysin-1 acts as a natural mammary tumor promoter." Oncogene 19(8): 1102-13. Stetler-Stevenson, W. G. (1996). "Dynamics of matrix turnover during pathologic remodeling of the extracellular matrix." Am J Pathol 148(5): 1345-50. Stout, R. D. (1993). "Macrophage activation by T cells: cognate and non-cognate signals." Curr Opin Immunol 5(3): 398-403. Suda, T., N. Takahashi, et al. (1999). "Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families." Endocr Rev 20(3): 345-57. Sundaram, K., R. Nishimura, et al. (2007). "RANK ligand signaling modulates the matrix metalloproteinase-9 gene expression during osteoclast differentiation." Exp Cell Res 313(1): 168-78. Surh, Y. J., S. S. Han, et al. (2000). "Inhibitory effects of curcumin and capsaicin on phorbol ester-induced activation of eukaryotic transcription factors, NF-kappaB and AP-1." Biofactors 12(1-4): 107-12. Svensson, L., K. S. Nandakumar, et al. (2002). "IL-4-deficient mice develop less acute but more chronic relapsing collagen-induced arthritis." Eur J Immunol 32(10): 2944-53. Swaminathan, R. (2001). "Biochemical markers of bone turnover." Clin Chim Acta 313(1-2): 95-105. Taams, L. S., J. M. van Amelsfort, et al. (2005). "Modulation of monocyte/macrophage function by human CD4+CD25+ regulatory T cells." Hum Immunol 66(3): 222-30. Takayanagi, H. (2005). "Inflammatory bone destruction and osteoimmunology." J Periodontal Res 40(4): 287-93. Takayanagi, H. (2007). "Osteoimmunology: shared mechanisms and crosstalk between the immune and bone systems." Nat Rev Immunol 7(4): 292-304. Thorbecke, G. J., R. Schwarcz, et al. (1999). "Modulation by cytokines of induction of oral tolerance to type II collagen." Arthritis Rheum 42(1): 110-8. Udagawa, N., N. Takahashi, et al. (1990). "Origin of osteoclasts: mature monocytes and macrophages are capable of differentiating into osteoclasts under a suitable microenvironment prepared by bone marrow-derived stromal cells." Proc Natl Acad Sci U S A 87(18): 7260-4. van Schaik, S. M. (2007). "Do pediatric patients with septic shock benefit from steroid therapy? A critical appraisal of "Low-dose hydrocortisone improves shock reversal and reduces cytokine levels in early hyperdynamic septic shock" by Oppert et al. (Crit Care Med 2005; 33:2457-2464)." Pediatr Crit Care Med 8(2): 174-6. VanSaun, M. N. and L. M. Matrisian (2006). "Matrix metalloproteinases and cellular motility in development and disease." Birth Defects Res C Embryo Today 78(1): 69-79. Walsh, N. C., M. Cahill, et al. (2003). "Multiple tissue-specific promoters control expression of the murine tartrate-resistant acid phosphatase gene." Gene 307: 111-23. Wang, H. H., H. L. Hsieh, et al. (2009). "Oxidized low-density lipoprotein induces matrix metalloproteinase-9 expression via a p42/p44 and JNK-dependent AP-1 pathway in brain astrocytes." Glia 57(1): 24-38. Wu, J. B., H. R. Chuang, et al. (2010). "A standardized aqueous extract of Anoectochilus formosanus ameliorated thioacetamide-induced liver fibrosis in mice: the role of Kupffer cells." Biosci Biotechnol Biochem 74(4): 781-7. Xie, F., C. F. Wu, et al. (2005). "The osteoprotective effect of Herba epimedii (HEP) extract in vivo and in vitro." Evid Based Complement Alternat Med 2(3): 353-61. Yagi, M., T. Miyamoto, et al. (2005). "DC-STAMP is essential for cell-cell fusion in osteoclasts and foreign body giant cells." J Exp Med 202(3): 345-51. Yoshimura, A. (2009). "Regulation of cytokine signaling by the SOCS and Spred family proteins." Keio J Med 58(2): 73-83. Yu, M., J. L. Moreno, et al. (2009). "Complex regulation of tartrate-resistant acid phosphatase (TRAP) expression by interleukin 4 (IL-4): IL-4 indirectly suppresses receptor activator of NF-kappaB ligand (RANKL)-mediated TRAP expression but modestly induces its expression directly." J Biol Chem 284(47): 32968-79. Zhang, Y., J. Cai, et al. (2007). "Antihyperglycemic activity of kinsenoside, a high yielding constituent from Anoectochilus roxburghii in streptozotocin diabetic rats." J Ethnopharmacol 114(2): 141-5. Zhao, Q., X. Wang, et al. (2010). "NFATc1: functions in osteoclasts." Int J Biochem Cell Biol 42(5): 576-9. Zhu, C. and J. F. Woessner, Jr. (1991). "A tissue inhibitor of metalloproteinases and alpha-macroglobulins in the ovulating rat ovary: possible regulators of collagen matrix breakdown." Biol Reprod 45(2): 334-42. 甘偉松 (1965). "台灣植物藥材誌." 第二輯: 86. 甘偉松 (1986). "藥用植物學." 藥用植物學: 647. 周先樂、陳世明、楊政民 (1975). "中藥藥理作用之研究." 臺灣醫誌 74: 702-707. 周佰隆 (2004). "台灣特有「藥王」台灣金線連." 中華民國自然生態保育協會大自然雜誌. 林文川、施純青、吳岳文 (2000). "妊娠期及授乳期母鼠投予臺灣金線連水抽取物對新生兒的影響." 中醫藥雜誌 11(2): 79-85. 林文川等 (2000). "臺灣金線連水抽取物對大鼠90天餵食毒性作用." 中醫藥雜誌 11(1): 19-29. 林俊義 (1986). "金線連的繁殖與推廣." 農友 37: 4-17. 梁文俐等 (1990). "臺灣產金線連之研究I.金線連之生理活性研究." 臺灣科學 43: 47-58. 劉新裕 (1998). "金線連之品種特性與毒性研究." 中華農業研究 47(3): 242-258. 蔡佳慈 (2007). "金線連糖苷對四氯化碳誘導之小鼠肝損傷的保護作用:經由抑制庫氏細胞的活化." 蔡新聲 (2003). "台灣金線連風華再現." 科學發展 364: 16-21. 蘇洪傑 (1975). "台灣野生蘭." 豐年叢書: 79-81.
摘要: 
台灣金線連是台灣傳統且珍貴草藥而金線連糖苷為主要的活性成分。在我們先前的實驗當中發現,金線連糖苷可以抑制肝臟的巨噬細胞也就是Kupffer cells經過LPS刺激所產生的TNF-α。這些的結果我們猜測了金線連糖苷可能可以抑制了巨噬細胞的活化。因為巨噬細胞都會參予各種的發炎過程。因此我們在這裡去探討了金線連糖苷在一些疾病當中巨噬細胞參予的影響包含了內毒素休克、關節炎跟骨質疏鬆。我們獲得了以下的結果:
1. 在LPS刺激之下,金線連糖苷可以明顯的抑制腹腔巨噬細胞釋放發炎物質包含了NO、TNF-α、IL-1β、MCP-1跟MIF。抗發炎細胞激素IL-10卻是有增加的現象。在動物模式中,預防跟治療的處理方式證明了金線連糖苷可以增加ICR小鼠在LPS的致死劑量 (80毫克/公斤,腹腔注射)的存活率。LPS誘導急性發炎的模式當中,金線連糖苷組別在1小時內的時候可以降低血清中TNF-α、IL-1β和IL-10和MCP-1和MIF的濃度。在注射後 24小時內之後金線連糖苷可以促進血清中IL-10濃度增加。總之金線連糖苷可以抑制前趨發炎細胞激素的產生和增強抗發炎細胞激素的生成。
2. 在關節炎動物模式當中,服用金線連糖苷可以抑制脾臟細胞TNF-α,IFN-γ跟IL-17分泌,卻可以增強IL-4跟IL-10的表現。我們利用RT-PCR發現,金線連糖苷也減少TNF-α、IL-1β和MMP-9的表現,同時增加了IL-10的表現。我們發現金線連糖苷也可以增加CD4+ CD25+調節性T細胞的數目。在H&E染色、TRAP還有免疫組織化學染色顯示用金線連糖苷餵食後細胞浸潤明顯減少且減少骨質的破壞。
3. μCT分析之後,金線連糖苷可以減少去卵巢母鼠所造成的骨質流失。金線連糖苷也減少了血漿C-末端肽Ⅰ型膠原蛋白的濃度,但並沒有抑制血漿鹼性磷酸酶活性。RT-PCR分析還說明金線連糖苷降低了脛骨當中TRAP跟MMP-9的表現。金線連糖苷是透過抑制在NF-κB調控路徑並且抑制NFATc1的表現進而去減少蝕骨細胞的特定基因的表現,如TRAP、DC-STAMP、MMP-9和cathepsin K。
4. MMP-9在疾病方面扮演著一個重要的角色。更進一步發現了金線連糖苷可以抑制MMP-9的表現。RT-PCR和Western blot的分析說明了,經過RANKL的處理 Raw 264.7細胞在金線連糖苷可以明顯抑制MMP-9基因和蛋白的表現。在MMP-9的啟動子活性當中也發現,金線連糖苷可以明顯減少冷光酵素表現。這些的結果可以證明,在RANKL刺激的Raw 264.7細胞當中金線連糖苷能夠有效地抑制MMP-9的產生是透過抑制p38/AP-1信號和NF-κB和ROS。
最後,我們證明了金線連糖苷在小鼠當中內毒素休克、關節炎還有骨質疏鬆動物模式的影響。金線連糖苷包含了多種的機轉去改善了一些症狀。首先金線連糖苷可以藉由抑制NF-κB去調控發炎物質的釋放。再來就是金線連糖苷可以透過MAPK跟ROS的路徑去抑制了MMP-9的產生。

Kinsenoside is a significant and active compound of the Anoectochilus formosanus (Orchidaceae), an important ethnomedicinal plant of Taiwan. In our previous works, we have reported kinsenoside inhibited the TNF-α secretion from Kupffer cells, hepatic macrophages, to LPS stimulation. These results suggest that kinsenoside may inhibit the activation of macrophages. Because macrophages participate in the various inflammatory process. Therefore in this study, we examined the effect of kinsenoside on the disease that including the participation of macrophages, sucsh as entotoxin shock, arthritis and osteoporosis. The following results were obtained.
1. In LPS-stimulated MPLMs, kinsenoside inhibited the inflammatory mediators, such as NO, TNF-α, IL-1β, MCP-1 and MIF production. In contrast, it stimulated anti-inflammatory cytokine IL-10 generation. In an animal model, both pre- and post-treatment of kinsenoside increased the survival rate of ICR mice challenged by LPS (80 mg/kg, i.p.). Pretreatment with kinsenoside decreased serum levels of TNF-α, IL-1β, IL-10, MCP-1 and MIF at 1 h after sublethal dose of LPS (40 mg/kg, i.p.) in mice. In contrast, kinsenoside enhanced serum IL-10 level at 24 h after LPS injection in mice. Kinsenoside inhibited the production of inflammatory mediators and enhanced anti-inflammatory cytokine generation.
2. In CIA mice, administration of kinsenoside significantly suppressed levels of TNF-α, IFN-γ and IL-17, but increased concentrations of IL-4 and IL-10 in the supernatants of each of the splenocytes. In RT-PCR analysis of mRNA expression in hind paws of CIA mice, kinsenoside decreased TNF-α, IL-1, and MMP-9 expression, and increased the expression of IL-10. H&E, TRAP and immunohistochemical staining indicated kinsenoside significantly reduced cellular infiltration and bone destruction. We also showed that kinsenoside enhanced the cell nimbers of CD4+ CD25+ Treg cells of splenocytes.
3. Microtomography scanning found that kinsenoside suppressed bone loss in OVX mice. Kinsenoside also decreased plasma CTx concentration but did not inhibit plasma ALP activities. RT-PCR analysis also showed that kinsenoside decreased the tibial mRNA expression of TRAP and MMP-9. In in vitro study, kinsenoside could inhibit RANKL-induced osteoclastogenesis in Raw 264.7 cells. Kinsenoside inhibited not only the RANKL-triggered nuclear translocations of NF-κB and NFATc1 but also the subsequent NFATc1 induction. NFATc1 promoted osteoclast-specific genes such as TRAP, DC-STAMP, and cathepsin K expression were inhibited by kinsenoside.
4. Since MMP-9 play an important role in several diseases. We further investigated the mechanism of kinsenoside to inhibit MMP-9 production. RT-PCR and Western blot analysis showed that kinsenoside significantly suppressed the mRNA and protein expression of inducible MMP-9 in RANKL-treated Raw 264.7 cells. Luciferase activity assay also showed that MMP-9 promoter activity was reduced by kinsenoside. Our data also showed that kinsenoside attenuated MMP-9 production via the inhibition of p38/AP-1 signaling, NF-κB and ROS activation in RANKL-treated Raw 264.7 cells.
In conclusion, our results demonstrated the beneficial effects of kinsenoside on endotoxic shock, CIA and osteoporosis in mice. Multiple mechanisms were involved in the ameliorative action of kinsenoside. Firstly, kinsenoside inhibited inflammatory mediators release via NF-κB pathway. Secondary, kinsenoside suppressed the MMP-9 secretion might be mediated through inhibition of MAPK pathway and ROS production.
URI: http://hdl.handle.net/11455/23172
其他識別: U0005-3101201122560300
Appears in Collections:生命科學系所

Show full item record
 

Google ScholarTM

Check


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.