Genistein stimulates the osteoblastic differentiation via NO/cGMP in bone marrow culture

• Published in: Journal of cellular biochemistry 2005-02, Vol.94 (2), p.307-316
• Main Authors: Pan, Wei, Quarles, L. Darryl, Song, Li-Hua, Yu, Yan-Hui, Jiao, Chen, Tang, Hong-Bo, Jiang, Chang-Hong, Deng, Han-Wu, Li, Yuan-Jian, Zhou, Hong-Hao, Xiao, Zhou-Sheng
• Format: Article
• Language: English
• Subjects: Alkaline Phosphatase - metabolism; Animals; Antineoplastic Agents - pharmacology; Bone Marrow Cells - cytology; Bone Marrow Cells - drug effects; Bone Marrow Cells - metabolism; Calcium - metabolism; Cell Differentiation; Cell Proliferation - drug effects; Core Binding Factor Alpha 1 Subunit; cyclic GMP; Cyclic GMP - metabolism; DNA-Binding Proteins - metabolism; Enzyme Inhibitors - pharmacology; Estradiol - analogs & derivatives; Estradiol - pharmacology; Estrogen Antagonists - pharmacology; estrogen receptor; Female; Fulvestrant; genistein; Genistein - pharmacology; Guanylate Cyclase - metabolism; mesenchymal stem cell; Mesenchymal Stem Cells - cytology; Mesenchymal Stem Cells - drug effects; Mesenchymal Stem Cells - metabolism; Mice; NG-Nitroarginine Methyl Ester - pharmacology; nitric oxide; Nitric Oxide - metabolism; Nitric Oxide Synthase - antagonists & inhibitors; Nitric Oxide Synthase - metabolism; osteoblast; Osteoblasts - drug effects; Osteoblasts - metabolism; phytoestrogen; Receptors, Estrogen - antagonists & inhibitors; Receptors, Estrogen - metabolism; Thymidine - metabolism; Transcription Factor AP-2; Transcription Factors - metabolism
• ISSN: 0730-2312; 1097-4644
• DOI: 10.1002/jcb.20308

The soybean phytoestrogen, genistein (Gen), has anabolic effects on bone through mechanisms that remain to be elucidated. We examined the role of nitric oxide (NO) and its downstream effector guanylyl cyclase (GC) in mediating the effects of Gen on the proliferation and osteoblastic maturation of primary mouse bone marrow‐derived mesenchymal stem cells (BMSCs). Gen (10−8 ∼ 10−6 M) resulted in a dose‐dependent increase in cell proliferation as measured by increased [3H]thymidine incorporation, and stimulated osteoblastic maturation as assessed by culture duration‐dependent increments in alkaline phosphatase (ALP) activity, calcium deposition into extracellular matrix and Runx2/Cbfa1 gene expression in BMSCs cultures. Gen also resulted in a dose‐dependent increase in NO synthase (NOS) activity, NO formation, and cGMP production in BMSCs cultures. The effects of Gen were mimicked by 17β‐estradiol (E2, 10−8 M). Concurrent treatment with the estrogen receptor (ER) antagonist ICI182,780 (10−7 M) or the NOS inhibitor L‐NAME (3 × 10−3 M) diminished the Gen (10−6 M)‐mediated increase in NOS activity, NO production, and cGMP content. In contrast, a soluble GC inhibitor 1H‐[1,2,4]oxadiazolo [4,3,‐a]quinoxalin‐1‐one (ODQ, 10−6 M) selectively blocked the Gen (10−6 M)‐mediated increase in cGMP content but not in NO production and NOS activity. Moreover, inhibition of ER, NOS activity or cGMP blocked Gen‐induced proliferation and osteoblastic differentiation of BMSCs and Runx2/Cbfa1 gene expression in culture. Gen has estrogen‐like activity and stimulates the proliferation and osteoblastic differentiation of mouse BMSCs at least in part through NO/cGMP pathway. © 2004 Wiley‐Liss, Inc.