Regulation by AMP-activated protein kinase of PGE2-induced osteoprotegerin synthesis in osteoblasts

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

doi:10.3892/mmr.2016.4900

Regulation by AMP-activated protein kinase of PGE2-induced osteoprotegerin synthesis in osteoblasts

Authors:
- Shingo Kainuma
- Takanobu Otsuka
- Gen Kuroyanagi
- Naohiro Yamamoto
- Rie Matsushima‑Nishiwaki
- Osamu Kozawa
- Haruhiko Tokuda
View Affiliations

Affiliations: Department of Orthopedic Surgery, Nagoya University Graduate School of Medical Sciences, Nagoya, Aichi 467‑8601, Japan, Department of Pharmacology, Gifu University Graduate School of Medicine, Gifu, Gifu 501‑1194, Japan
Published online on: February 17, 2016 https://doi.org/10.3892/mmr.2016.4900
Pages: 3363-3369

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Adenosine monophosphate-activated protein kinase (AMPK) is currently recognized to act as a key sensing enzyme in the regulation of cellular energy homeostasis. It has been previously demonstrated that prostaglandin E2 (PGE2) stimulates the synthesis of osteoprotegerin (OPG) through the activation of p38 mitogen-activated protein (MAP) kinase, p44/p42 MAP kinase and stress-activated protein kinase/c‑Jun N‑terminal kinase (SAPK/JNK) in osteoblast‑like MC3T3‑E1 cells. In the present study, it was investigated whether AMPK is implicated in the PGE2‑induced OPG synthesis in MC3T3‑E1 cells. PGE2 was observed to induce the phosphorylation of AMPKα (Thr‑172) and AMPKβ (Ser‑108) in a time‑dependent manner. PGE2 additionally induced the phosphorylation of acetyl‑coenzyme A (CoA) carboxylase, a direct substrate of AMPK. Compound C, an inhibitor of AMPK, which attenuated the phosphorylation of acetyl‑CoA carboxylase, significantly suppressed the PGE2‑stimulated OPG release and the mRNA expression level. Compound C failed to affect the PGE2‑stimulated phosphorylation of p38 MAP kinase or p44/p42 MAP kinase. On the contrary, the phosphorylation of SAPK/JNK was markedly attenuated by compound C. The results of the current study suggest that AMPK acts as a positive regulator in PGE2-stimulated OPG synthesis via SAPK/JNK signaling in osteoblasts.

View Figures

View References

1	Fogarty S and Hardie DG: Development of protein kinase activators: AMPK as a target in metabolic disorders and cancer. Biochim Biophys Acta. 1804:581–591. 2010. View Article : Google Scholar
2	Mihaylova MM and Shaw RJ: The AMPK signalling pathway coordinates cell growth, autophagy and metabolism. Nat Cell Biol. 13:1016–1023. 2011. View Article : Google Scholar : PubMed/NCBI
3	Rutter GA and Leclerc I: The AMP-regulated kinase family: Enigmatic targets for diabetes therapy. Mol Cell Endocrinol. 297:41–49. 2009. View Article : Google Scholar
4	Karsenty G and Wagner EF: Reaching a genetic and molecular understanding of skeletal development. Dev Cell. 2:389–406. 2002. View Article : Google Scholar : PubMed/NCBI
5	Shah M, Kola B, Bataveljic A, Arnett TR, Viollet B, Saxon L, Korbonits M and Chenu C: AMP-activated protein kinase (AMPK) activation regulates in vitro bone formation and bone mass. Bone. 47:309–319. 2010. View Article : Google Scholar : PubMed/NCBI
6	Kato K, Tokuda H, Adachi S, Matsushima-Nishiwaki R, Natsume H, Yamakawa K, Gu Y, Otsuka T and Kozawa O: AMP-activated protein kinase positively regulates FGF-2-stimulated VEGF synthesis in osteoblasts. Biochem Biophys Res Commun. 400:123–127. 2010. View Article : Google Scholar : PubMed/NCBI
7	Simonet WS, Lacey DL, Dunstan CR, Kelley M, Chang MS, Lüthy R, Nguyen HQ, Wooden S, Bennett L, Boone T, et al: Osteoprotegerin: A novel secreted protein involved in the regulation of bone density. Cell. 89:309–319. 1997. View Article : Google Scholar : PubMed/NCBI
8	Kong YY, Yoshida H, Sarosi I, Tan HL, Timms E, Capparelli C, Morony S, Oliveira-dos-Santos AJ, Van G, Itie A, et al: OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. Nature. 397:315–323. 1999. View Article : Google Scholar : PubMed/NCBI
9	Theoleyre S, Wittrant Y, Tat SK, Fortun Y, Redini F and Heymann D: The molecular triad OPG/RANK/RANKL: Involvement in the orchestration of pathophysiological bone remodeling. Cytokine Growth Factor Rev. 15:457–475. 2004. View Article : Google Scholar : PubMed/NCBI
10	Hikiji H, Takato T, Shimizu T and Ishii S: The roles of prostanoids, leukotrienes and platelet-activating factor in bone metabolism and disease. Prog Lipid Res. 47:107–126. 2008. View Article : Google Scholar : PubMed/NCBI
11	Zhu Z, Fu C, Li X, Song Y, Li C, Zou M, Guan Y and Zhu Y: Prostaglandin E₂ promotes endothelial differentiation from bone marrow-derived cells through AMPK activation. PloS One. 6:e235542011. View Article : Google Scholar
12	Yamamoto N, Tokuda H, Kuroyanagi G, Mizutani J, Matsushima-Nishiwaki R, Kondo A, Kozawa O and Otsuka T: Regulation by resveratrol of prostaglandin E₂-stimulated osteoprotegerin synthesis in osteoblasts. Int J Mol Med. 34:1439–1445. 2014.PubMed/NCBI
13	Sudo H, Kodama HA, Amagai Y, Yamamoto S and Kasai S: In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria. J Cell Biol. 96:191–198. 1983. View Article : Google Scholar : PubMed/NCBI
14	Kozawa O, Tokuda H, Miwa M, Kotoyori J and Oiso Y: Cross-talk regulation between cyclic AMP production and phosphoinositide hydrolysis induced by prostaglandin E₂ in osteoblast-like cells. Exp Cell Res. 198:130–134. 1992. View Article : Google Scholar : PubMed/NCBI
15	Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227:680–685. 1970. View Article : Google Scholar : PubMed/NCBI
16	Kato K, Ito H, Hasegawa K, Inaguma Y, Kozawa O and Asano T: Modulation of the stress-induced synthesis of hsp27 and alpha B-crystallin by cyclic AMP in C6 rat glioma cells. J Neurochem. 66:946–950. 1996. View Article : Google Scholar : PubMed/NCBI
17	Hawley SA, Davison M, Woods A, Davies SP, Beri RK, Carling D and Hardie DG: Characterization of the AMP-activated protein kinase kinase from rat liver and identification of threonine 172 as the major site at which it phosphorylates AMP-activated protein kinase. J Biol Chem. 271:27879–27887. 1996. View Article : Google Scholar : PubMed/NCBI
18	Zhou G, Myers R, Li Y, Chen Y, Shen X, Fenyk-Melody J, Wu M, Ventre J, Doebber T, Fujii N, et al: Role of AMP-activated protein kinase in mechanism of metformin action. J Clin Invest. 108:1167–1174. 2001. View Article : Google Scholar : PubMed/NCBI
19	Kyriakis JM and Avruch J: Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. Physiol Rev. 81:807–869. 2001.PubMed/NCBI
20	Widmann C, Gibson S, Jarpe MB and Johnson GL: Mitogen-activated protein kinase: Conservation of a three-kinase module from yeast to human. Physiol Rev. 79:143–180. 1999.PubMed/NCBI
21	Kanazawa I, Yamaguchi T, Yano S, Yamauchi M and Sugimoto T: Metformin enhances the differentiation and mineralization of osteoblastic MC3T3-E1 cells via AMP kinase activation as well as eNOS and BMP-2 expression. Biochem Biophys Res Commun. 375:414–419. 2008. View Article : Google Scholar : PubMed/NCBI
22	Kim JE, Ahn MW, Baek SH, Lee IK, Kim YW, Kim JY, Dan JM and Park SY: AMPK activator, AICAR, inhibits palmitate-induced apoptosis in osteoblast. Bone. 43:394–404. 2008. View Article : Google Scholar : PubMed/NCBI
23	Jang WG, Kim EJ, Lee KN, Son HJ and Koh JT: AMP-activated protein kinase (AMPK) positively regulates osteoblast differentiation via induction of Dlx5-dependent Runx2 expression in MC3T3E1 cells. Biochem Biophys Res Commun. 404:1004–1009. 2011. View Article : Google Scholar