Biochanin A Promotes Osteogenic but Inhibits Adipogenic Differentiation: Evidence with Primary Adipose-Derived Stem Cells

Evidence-Based Complementary and Alternative Medicine, Jun 2013

Biochanin A has promising effects on bone formation in vivo, although the underlying mechanism remains unclear yet. This study therefore aimed to investigate whether biochanin A regulates osteogenic and adipogenic differentiation using primary adipose-derived stem cells. The effects of biochanin A (at a physiologically relevant concentration of 0.1–1 M) were assessed in vitro using various approaches, including Oil red O staining, Nile red staining, alizarin red S staining, alkaline phosphatase (ALP) activity, flow cytometry, RT-PCR, and western blotting. The results showed that biochanin A significantly suppressed adipocyte differentiation, as demonstrated by the inhibition of cytoplasmic lipid droplet accumulation, along with the inhibition of peroxisome proliferator-activated receptor gamma (PPARγ), lipoprotein lipase (LPL), and leptin and osteopontin (OPN) mRNA expression, in a dose-dependent manner. On the other hand, treatment of cells with 0.3 M biochanin A increased the mineralization and ALP activity, and stimulated the expression of the osteogenic marker genes ALP and osteocalcin (OCN). Furthermore, biochanin A induced the expression of runt-related transcription factor 2 (Runx2), osteoprotegerin (OPG), and Ras homolog gene family, member A (RhoA) proteins. These observations suggest that biochanin A prevents adipogenesis, enhances osteoblast differentiation in mesenchymal stem cells, and has beneficial regulatory effects in bone formation.

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Biochanin A Promotes Osteogenic but Inhibits Adipogenic Differentiation: Evidence with Primary Adipose-Derived Stem Cells

Biochanin A Promotes Osteogenic but Inhibits Adipogenic Differentiation: Evidence with Primary Adipose-Derived Stem Cells Shu-Jem Su,1 Yao-Tsung Yeh,1 Shu-Hui Su,2 Kee-Lung Chang,3 Huey-Wen Shyu,1 Kuan-Ming Chen,3 and Hua Yeh1 1Department of Medical Laboratory Science and Biotechnology, School of Medicine and Health Sciences, FooYin University, 151 Chin-Hsueh Road, Ta-Liao, Kaohsiung 83101, Taiwan 2Institute of Medical Sciences, College of Medicine, Tzu Chi University, Hualien, Taiwan 3Department of Biochemistry, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan Received 29 March 2013; Accepted 28 May 2013 Academic Editor: Stephen Cho Wing Sze Copyright © 2013 Shu-Jem Su et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Biochanin A has promising effects on bone formation in vivo, although the underlying mechanism remains unclear yet. This study therefore aimed to investigate whether biochanin A regulates osteogenic and adipogenic differentiation using primary adipose-derived stem cells. The effects of biochanin A (at a physiologically relevant concentration of 0.1–1  M) were assessed in vitro using various approaches, including Oil red O staining, Nile red staining, alizarin red S staining, alkaline phosphatase (ALP) activity, flow cytometry, RT-PCR, and western blotting. The results showed that biochanin A significantly suppressed adipocyte differentiation, as demonstrated by the inhibition of cytoplasmic lipid droplet accumulation, along with the inhibition of peroxisome proliferator-activated receptor gamma (PPARγ), lipoprotein lipase (LPL), and leptin and osteopontin (OPN) mRNA expression, in a dose-dependent manner. On the other hand, treatment of cells with 0.3  M biochanin A increased the mineralization and ALP activity, and stimulated the expression of the osteogenic marker genes ALP and osteocalcin (OCN). Furthermore, biochanin A induced the expression of runt-related transcription factor 2 (Runx2), osteoprotegerin (OPG), and Ras homolog gene family, member A (RhoA) proteins. These observations suggest that biochanin A prevents adipogenesis, enhances osteoblast differentiation in mesenchymal stem cells, and has beneficial regulatory effects in bone formation. 1. Introduction Menopause or age-related osteoporosis is associated with a progressive decrease in bone formation and an increase in adipogenesis in the bone marrow, thus increasing the risk of bone fractures [1, 2]. Bone development and homeostasis is mainly determined by the biological balance between osteogenesis and adipogenesis [3]. Recent studies in osteoporotic patients observed abnormal bone marrow-derived stem cells (BMSCs), wherein the BMSCs were deficient in their ability to differentiate into an osteogenic lineage and displayed increased adipogenic potential [4, 5]. Adipose-derived stem cells (ADSCs) and BMSCs have been most extensively evaluated as they offer the most accessible source of MSCs for use in research and clinical applications. They also display similar differentiation potentials. However, ADSCs may have greater advantage than BMSCs in clinical uses due to the following reasons. First, ADSCs are relatively abundant and easy to isolate. Second, ADSCs have been shown to have multilineage potential capable of differentiating into adipocyte, chondrocyte, and osteoblast, and they share similar phenotypic and functional characteristics with the BMSCs [6, 7]. Third, MSCs extracted from adipose tissue are less invasive and less expensive than those extracted from bone marrow [8]. Fourth, ADSCs have a significantly shorter doubling time [9]. Fifth, few or no ethical issue is involved in using ADSCs for research and medical purposes. Collectively, these intrinsic characteristics and advantages make ADSCs an ideal stem cell source for cell-based tissue engineering and/or therapies [7, 10]. Additionally, ADSCs were used in the regenerative treatment of traumatic calvarial bone defects in humans [11]. ADSCs can differentiate into multiple mesenchymal lineages, and they are a unique model to improve our understanding of the early differentiation events. It was reported that inhibiting marrow adipogenesis could help to either prevent or treat osteopenic disorders [12]. Therefore, more functional bone cells can be generated by the inhibition of marrow adipogenesis, with a concomitant increase in osteogenesis; this could either prevent further increases in adipocyte formation or divert existing adipocytes to become more osteoblastic, resulting in increased functional bone cells [13]. The differentiation of adipocytes requires activation by several adipogenesis-related genes, including peroxisome proliferator-activated receptor gamma (PPARγ) and the PPARγ-target gene lipoprotein lipase (LPL) [14]. PPARγ is a key transcr (...truncated)


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Shu-Jem Su, Yao-Tsung Yeh, Shu-Hui Su, Kee-Lung Chang, Huey-Wen Shyu, Kuan-Ming Chen, Hua Yeh. Biochanin A Promotes Osteogenic but Inhibits Adipogenic Differentiation: Evidence with Primary Adipose-Derived Stem Cells, Evidence-Based Complementary and Alternative Medicine, 2013, 2013, DOI: 10.1155/2013/846039