Involvement of AMP-activated protein kinase in TGF-β-stimulated VEGF synthesis in osteoblasts

Mizutani,Jun; Tokuda,Haruhiko; Matsushima-Nishiwaki,Rie; Kato,Kenji; Kondo,Akira; Natsume,Hideo; Kozawa,Osamu; Otsuka,Takanobu

doi:10.3892/ijmm.2012.893

Involvement of AMP-activated protein kinase in TGF-β-stimulated VEGF synthesis in osteoblasts

Authors:
- Jun Mizutani
- Haruhiko Tokuda
- Rie Matsushima-Nishiwaki
- Kenji Kato
- Akira Kondo
- Hideo Natsume
- Osamu Kozawa
- Takanobu Otsuka
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Affiliations: Department of Orthopedic Surgery, Nagoya City University Graduate School of Medical Sciences, Mizuho-Cho, Mizuho-Ku, Nagoya 467-8601, Japan, Department of Pharmacology, Gifu University Graduate School of Medicine, Gifu 501-1194, Japan
Published online on: January 23, 2012 https://doi.org/10.3892/ijmm.2012.893
Pages: 550-556
Copyright: © Mizutani et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

It is generally recognized that AMP-activated protein kinase (AMPK) acts as a key regulator of energy homeostasis. We have previously shown that transforming growth factor-β (TGF-β) stimulates synthesis of vascular endothelial growth factor (VEGF) via p44/p42 mitogen-activated protein (MAP) kinase, stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) and p38 MAP kinase in osteoblast-like MC3T3-E1 cells. In the present study, we investigated whether AMPK is involved in the TGF-β-stimulated VEGF synthesis in osteoblast-like MC3T3-E1 cells. TGF-β time-dependently induced the phosphorylation of the AMPK α-subunit (Thr172) and the AMPK β-subunit (Ser108). Compound C, an AMPK inhibitor, significantly reduced the TGF-β-stimulated VEGF release. The inhibitory effect of compound C was also observed in normal human osteoblasts (NHOst). Although compound C failed to affect the TGF-β-induced phosphorylation of SAPK/JNK, p38 MAP kinase or Smad2, it markedly suppressed the TGF-β-induced phosphorylation of both MEK1/2 and p44/p42 MAP kinase. In addition, compound C significantly suppressed the VEGF mRNA expression induced by TGF-β. Taken together, our results strongly suggest that AMPK is involved in TGF-β-stimulated VEGF synthesis, and that it functions at a point upstream of MEK1/2.

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