秦皮素促進成骨細胞分化以及阻斷發炎性細胞激素所導致成骨細胞凋亡之作用機制

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Title:	秦皮素促進成骨細胞分化以及阻斷發炎性細胞激素所導致成骨細胞凋亡之作用機制 Fraxetin stimulates differentiation and inhibits the induction of inflammatory cytokines-mediated apoptosis in human osteoblasts
Authors:	黃郁婷 Yu-ting Huang
Contributors:	張竣凱郭柏麟嘉南藥理科技大學：生物科技研究所
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Inhibitory effect of glucocorticoid for osteoblast apoptosis induced by activated peripheral blood mononuclear cells. Endocrinology. 1998;139:2032-40. 71.Hengartner MO. The biochemistry of apoptosis. Nature. 2000;407:770-76. 72.Nagata S, Golstein P. The Fas death factor. Science. 1995;267:1449-56. 73.Bratton SB, MacFarlane M, Cain K, Cohen GM. Protein complexes activate distinct caspase cascades in death receptor and stress-induced apoptosis. Exp Cell Res. 2000;256:27-33. 74.Tran SE, Meinander A, Holmstrom TH, Rivero-Muller A, Heiskanen KM, Linnau EK, et al. Heat stress downregulates FLIP and sensitizes cells to Fas receptor-mediated apoptosis. Cell Death Differ. 2003;10:1137-47. 細胞分化鹼性磷酸酶骨鈣素成型蛋白2 骨質疏鬆症腫瘤壞死因子白介素-1 Fraxetin Osteoblasts Cell differentiation ALP
Date:	2006
Issue Date:	2008-11-24 17:01:32 (UTC+8)
Abstract:	秦皮素(7,8-dihydroxy-6-methoxy coumarin)，是香豆素(coumarin)的衍生物。本研究分析秦皮素對人類成骨細胞株的分化作用中顯示，秦皮素能增加ALP活性與osteocalcin的分泌。而推測秦皮素會促進早期以及晚期成骨細胞的分化。我們亦探討秦皮素對於BMP-2以及BMP-4誘導細胞分化作用的情形，研究顯示BMP-2或BMP-4與秦皮素併用時，對細胞株ALP活性以及osteocalcin的分泌並不具有加成作用。然而BMP的抑制劑Noggin，可明顯阻斷由BMP-2以及BMP-4與秦皮素所誘導的ALP活性與osteocalcin的分泌。由此顯示秦皮素誘導成骨細胞的成熟與分化乃透過增加BMP-2以及BMP-4的表現所致。因此，秦皮素可做為預防以及治療骨質疏鬆症的天然藥物。在發炎性關節腔疾病的病人，例如類風濕性關節炎的患者，成骨細胞的存活對於這些疾病引起骨質疏鬆症的形成扮演著決定性的因素。在本研究中乃分析秦皮素是否會影響發炎性細胞激素所誘導成骨細胞的凋亡。研究顯示TNF-ㄘ呰L-1β會加強經anti-Fas IgM處理之MG-63的細胞凋亡，但是，單獨給予TNF-ㄘ呰L-1β並不會引起成骨細胞的凋亡。秦皮素不僅會抑制anti-Fas IgM所誘導的細胞凋亡，也會阻斷Fas ligand與TNF-ㄘ呰L-1β誘導細胞凋亡的協同作用。在探討秦皮素抑制成骨細胞凋亡之作用機制方面，秦皮素主要是抑制由TNF-ㄘMIL-1β所調節之Fas表現活性，同時並促進FLIP的表現而抑制caspase-8、caspase-3的活化作用。實驗結果顯示秦皮素可藉由抑制發炎性細胞激素所導致成骨細胞的凋亡，而預防骨質疏鬆症。 Fraxetin (7,8-dihydroxy-6-methox coumarin), a coumarin derivative, was investigated for its effects on differentiation of osteoblasts. By means of alkaline phosphatase (ALP) activity and osteocalcin ELISA assay, we have shown that fraxetin exhibits a significant induction of differentiation in two human osteoblast-like cell lines, MG-63 and hFOB. Alkaline phosphatase and osteocalcin are phenotypic markers for early-stage differentiated osteoblasts and terminally differentiated osteoblasts, respectively. Our results indicated that fraxetin stimulated osteoblast differentiation at various stages (from osteoprogenitors to terminally differentiated osteoblasts). Induction of differentiation by fraxetin was associated with increased BMP-2 and BMP-4 productions. Addition of purified BMP-2 and BMP-4 proteins did not increase the upregulation of ALP activity and osteocalcin secretion by fraxetin, whereas the BMPs antagonist noggin blocked both fraxetin and BMP-2 and BMP-4 mediated ALP activity and osteocalcin secretion enhancement, indicating that BMP-2 and BMP-4 productions are required in fraxetin-mediated osteoblast maturation and differentiation. These findings are novel and may be important in the treatment and prevention of osteoporosis. The survival of osteoblast cells is one of the determinants of the development of osteoporosis in patients with inflamed synovium, such as in rheumatoid arthritis (RA). By means of alkaline phosphatase (ALP) activity and osteocalcin ELISA assay, this study have shown that fraxetin exhibits a significant induction of differentiation in the human osteoblast-like cell line MG-63. In addition, this study also assessed whether fraxetin affects inflammatory cytokine-mediated apoptosis in osteoblast cells. TNF-?or IL-1β enhance apoptotic DNA fragmentation in anti-Fas IgM-treated MG-63 cells by increasing Fas receptor expression. However, TNF-?or IL-1β treatment alone does not induce apoptosis. Treatment of MG-63 cells with fraxetin not only inhibited anti-Fas IgM-induced apoptosis, but also blocked the synergetic effect of anti-Fas IgM with TNF-?or IL-1β on cell death. The apoptotic inhibition of fraxetin is associated with inhibition of TNF-?and IL-1β-mediated Fas expression and enhancement of FLIP expression, resulting in a decrease of caspase-8 and caspase-3 activation. These results indicate a potential use of fraxetin in preventing osteoporosis by inhibiting inflammatory cytokine-mediated apoptosis in osteoblast cells.
Relation:	校內公開，校外永不公開
Appears in Collections:	[Dept. of Biotechnology (including master's program)] Dissertations and Theses

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