Possible involvement of AMP‑activated protein kinase in PGE1‑induced synthesis of osteoprotegerin in osteoblasts

Kainuma,Shingo; Otsuka,Takanobu; Kuroyanagi,Gen; Yamamoto,Naohiro; Matsushima‑Nishiwaki,Rie; Kozawa,Osamu; Tokuda,Haruhiko

doi:10.3892/etm.2016.3099

Possible involvement of AMP‑activated protein kinase in PGE1‑induced synthesis of osteoprotegerin in osteoblasts

Authors:
- Shingo Kainuma
- Takanobu Otsuka
- Gen Kuroyanagi
- Naohiro Yamamoto
- Rie Matsushima‑Nishiwaki
- Osamu Kozawa
- Haruhiko Tokuda
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Affiliations: Department of Orthopedic Surgery, Nagoya City University Graduate School of Medical Sciences, Nagoya, Aichi 467‑8601, Japan, Department of Pharmacology, Gifu University Graduate School of Medicine, Gifu 501‑1194, Japan
Published online on: February 22, 2016 https://doi.org/10.3892/etm.2016.3099
Pages: 2042-2048

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Abstract

AMP‑activated protein kinase (AMPK) is firmly established as a central regulator of cellular energy homeostasis. We have previously reported that prostaglandin E1 (PGE1) stimulates the synthesis of osteoprotegerin through p38 mitogen‑activated protein (MAP) kinase and stress‑activated protein kinase/c‑Jun N‑terminal kinase (SAPK/JNK) in osteoblast‑like MC3T3‑E1 cells. The present study investigated the involvement of AMPK in PGE1‑induced osteoprotegerin synthesis in MC3T3‑E1 cells. The levels of osteoprotegerin were measured using an enzyme‑linked immunosorbent assay, while the phosphorylation of AMPK, acetyl‑CoA carboxylase, p38 MAP kinase and SAPK/JNK were analyzed by western blotting. In addition, the mRNA expression levels of osteoprotegerin were determined by a reverse transcription‑quantitative polymerase chain reaction. It was revealed that PGE1 significantly induced the phosphorylation of the α and β subunits of AMPK in a time‑dependent manner (P<0.05). In addition, acetyl‑CoA carboxylase, a direct substrate of AMPK, was significantly phosphorylated by PGE1 (P<0.05). Compound C, an AMPK inhibitor, was revealed to suppress the phosphorylation of acetyl‑CoA carboxylase, which significantly reduced the release and mRNA expression levels of PGE1‑stimulated osteoprotegerin (P<0.05). However, the PGE1‑induced phosphorylation of p38 MAP kinase and SAPK/JNK were not affected by compound C. The results of the present study indicated that AMPK may positively regulate PGE1‑stimulated osteoprotegerin synthesis in osteoblasts; thus providing novel insight into the regulatory mechanisms underlying bone metabolism.

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