Sambucus Formosana Stimulates Differentiation of MC3T3-E1 Osteoblasts through BMP-2 and β-Catenin Pathways

有骨消(Sambucus formosana)藉由活化BMP-2及β-catenin訊息路徑刺激MC3T3-E1前驅造骨细胞分化

邱文慧(Wen-Fei Chiou) ； 陳建志(Chien-Chih Chen) ； 洪銘宏(Ming-Hung Hung)

《中醫藥雜誌》 21卷1&2期 (2010/07) Pp. 1-15

https://doi.org/10.6940/JCM.201007_21(1_2).01

摘要

臺灣民問草藥有骨消(Sambucus formosana, SF)常用於治療骨折，本實驗研究有骨消是否可促進MC3T3-E1前驅造骨細胞的分化及礦化，同時探討其對造骨新生(osteoblastogenesis)訊息路徑的影響。結果顯示有骨消乙醇萃取物之氯仿可溶層製備物(SF-C)呈現濃度相關性地增加鹼性磷酸嗨(ALP)活性、osteopontin濃度、osteocalcin蛋白及骨形態蛋白－2(BMP-2)表現、以及促進鈣化骨節的形成；同時SF-C促進造骨細胞分化的作用可被noggin、wortmannin、以及SB2003580所抑制，而SF-C也會增加磷酸化Smad1/5/8、磷酸化Akt、以及磷酸化P38的表現，猜測這些訊息分子均參與調控SF-C的造骨新生作用。另一方面，β-catenin在調控骨質生成作用上扮演相當重要的角色，本研究發現轉染β-catenin siRNA後可明顯抑制SF-C促進分化的能力。此外，雖然SF-C並不影響前驅造骨細胞分泌RANKL的能力，但卻會促進osteoprotegerin(OPG)的生成。總言之，我們首先證實了有骨消乙醇草取物之氣仿可溶層製備物(SF-C)確實會促進前驅造骨細胞的分化及礦化，而根據機轉探討結果發現SF-C的骨生成作用可能與活化BMP-2-及β-catenin之訊息路徑有關，再者，SF-C也可藉由提高OPG/RANKL的比值來問接抑制蝕骨細胞的分化及活化。以上實驗驗證了有骨消應用於改善骨折的臨床功效。

關鍵字

有骨消 ； 造骨細胞 ； 骨形態蛋白－2 BMP-2 ； β-catenin ； p38 ； Akt

並列摘要

Sambucus formosana Nakai is used as a folk medicine in Taiwan to remedy bone fracture. Here we examined the effects of S. formosana on the differentiation and mineralization of MC3T3-E1 osteoblasts. Then, the impact of S. formosana on signaling pathways known to be implicated in osteoblastogenesis was explored. Results showed the chloroform soluble fraction prepared from the ethanol extract of S. formosana (defined as SF-C) concentration-dependently increased in alkaline phosphatase (ALP) activity, levels of osteopontin, osteocalcin and bone morphogenetic proteins-2 (BMP-2), and calcium nodules formation. SF-C evoked differentiation was significantly abrogated by noggin, wortmannin and SB2003580 treatment. SF-C also induced Smad1/5/8, Akt and p38 phosphorylation suggesting that these signal pathways were probably involved. β-catenin plays a pivotal role in regulating bone mass formation. The results further showed that transfection with β-catenin siRNA significantly attenuated SF-C induced ALP activity. Considering the indirect effect on the regulation of osteoclastogenesis, we found SF-E could stimulate osteoprotegerin (OPG) production although have no effect on RANKL synthesis from MC3T3-E1 preosteoblasts. In summary, our results proved the clinical application of S. formosana and indicated that SF-C has bone anabolic effect being probably through BMP-2- and β-catenin-dependant signal pathways. Further, repression of bone catabolism via up-regulation of OPG/RANKL ratio may also participate in cure bone fracture by S. formosana.

並列關鍵字

Sambucus formosana ； osteoblast ； BMP-2 ； β-catenin ； p38 ； Akt