Incretins amplify TNF-α-stimulated IL-6 synthesis in osteoblasts: Suppression of the IκB/NF-κB pathway

Fujita,Kazuhiko; Tokuda,Haruhiko; Yamamoto,Naohiro; Kainuma,Shingo; Kawabata,Tetsu; Sakai,Go; Kuroyanagi,Gen; Matsushima‑Nishiwaki,Rie; Harada,Atsushi; Kozawa,Osamu; Otsuka,Takanobu

doi:10.3892/ijmm.2017.2892

Incretins amplify TNF-α-stimulated IL-6 synthesis in osteoblasts: Suppression of the IκB/NF-κB pathway

Authors:
- Kazuhiko Fujita
- Haruhiko Tokuda
- Naohiro Yamamoto
- Shingo Kainuma
- Tetsu Kawabata
- Go Sakai
- Gen Kuroyanagi
- Rie Matsushima‑Nishiwaki
- Atsushi Harada
- Osamu Kozawa
- Takanobu Otsuka
View Affiliations

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, Department of Orthopedic Surgery, National Center for Geriatrics and Gerontology, Obu, Aichi 474‑8511, Japan
Published online on: February 15, 2017 https://doi.org/10.3892/ijmm.2017.2892
Pages: 1053-1060

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

Incretins including glucagon-like peptide-1 (GLP-1) and glucose‑dependent insulinotropic polypeptide (GIP) secreted from the small intestine after oral food ingestion are currently recognized to stimulate insulin secretion from pancreatic β cells. We previously reported that p70 S6 kinase limits the tumor necrosis factor‑α (TNF‑α)‑stimulated interleukin-6 (IL‑6) synthesis in osteoblast‑like MC3T3‑E1 cells. In the present study, we investigated the effects of incretins on the TNF‑α‑induced IL‑6 synthesis and the underlying mechanism in MC3T3‑E1 cells. GLP‑1 and GIP significantly upregulated both TNF‑α‑stimulated IL‑6 release and mRNA levels. Wedelolactone, an inhibitor of IκB kinase, amplified the TNF-α-induced IL‑6 release. GLP‑1 significantly attenuated the TNF‑α‑induced phosphorylation of IκB without affecting the phosphorylation of p70 S6 kinase. On the other hand, GLP‑1 markedly induced the phosphorylation of cAMP response element-binding protein (CREB). H‑89, an inhibitor of protein kinase A, significantly suppressed the enhancement by GLP-1 of TNF-α-stimulated IL‑6 release. Dibutyryl cAMP, a permeable analogue of cAMP, which suppressed the TNF-α-induced IκB phosphorylation, amplified the IL‑6 release. These results strongly suggest that incretins upregulate the TNF-α-stimulated IL‑6 synthesis in osteoblasts, and that the amplifying effect of incretin is exerted via reducing the IκB/NF‑κB pathway through the adenylyl cyclase-cAMP system.

View Figures

View References

1	Kular J, Tickner J, Chim SM and Xu J: An overview of the regulation of bone remodelling at the cellular level. Clin Biochem. 45:863–873. 2012. View Article : Google Scholar : PubMed/NCBI
2	Chim SM, Tickner J, Chow ST, Kuek V, Guo B, Zhang G, Rosen V, Erber W and Xu J: Angiogenic factors in bone local environment. Cytokine Growth Factor Rev. 24:297–310. 2013. View Article : Google Scholar : PubMed/NCBI
3	Boyce BF and Xing L: Functions of RANKL/RANK/OPG in bone modeling and remodeling. Arch Biochem Biophys. 473:139–146. 2008. View Article : Google Scholar : PubMed/NCBI
4	Kwan Tat S, Padrines M, Théoleyre S, Heymann D and Fortun Y: IL-6, RANKL, TNF-alpha/IL-1: Interrelations in bone resorption pathophysiology. Cytokine Growth Factor Rev. 15:49–60. 2004. View Article : Google Scholar : PubMed/NCBI
5	Zou W, Hakim I, Tschoep K, Endres S and Bar-Shavit Z: Tumor necrosis factor-α mediates RANK ligand stimulation of osteoclast differentiation by an autocrine mechanism. J Cell Biochem. 83:70–83. 2001. View Article : Google Scholar : PubMed/NCBI
6	Gilbert L, He X, Farmer P, Boden S, Kozlowski M, Rubin J and Nanes MS: Inhibition of osteoblast differentiation by tumor necrosis factor-α. Endocrinology. 141:3956–3964. 2000.PubMed/NCBI
7	Puimège L, Libert C and Van Hauwermeiren F: Regulation and dysregulation of tumor necrosis factor receptor-1. Cytokine Growth Factor Rev. 25:285–300. 2014. View Article : Google Scholar : PubMed/NCBI
8	Kozawa O, Suzuki A, Kaida T, Tokuda H and Uematsu T: Tumor necrosis factor-alpha autoregulates interleukin-6 synthesis via activation of protein kinase C. Function of sphingosine 1-phosphate and phosphatidylcholine-specific phospholipase C. J Biol Chem. 272:25099–25104. 1997. View Article : Google Scholar : PubMed/NCBI
9	Minamitani C, Tokuda H, Adachi S, Matsushima-Nishiwaki R, Yamauchi J, Kato K, Natsume H, Mizutani J, Kozawa O and Otsuka T: p70 S6 kinase limits tumor necrosis factor-α-induced interleukin-6 synthesis in osteoblast-like cells. Mol Cell Endocrinol. 315:195–200. 2010. View Article : Google Scholar
10	Hirano T, Yasukawa K, Harada H, Taga T, Watanabe Y, Matsuda T, Kashiwamura S, Nakajima K, Koyama K, Iwamatsu A, et al: Complementary DNA for a novel human interleukin (BSF-2) that induces B lymphocytes to produce immunoglobulin. Nature. 324:73–76. 1986. View Article : Google Scholar : PubMed/NCBI
11	Johnson RW, Brennan HJ, Vrahnas C, Poulton IJ, McGregor NE, Standal T, Walker EC, Koh TT, Nguyen H, Walsh NC, et al: The primary function of gp130 signaling in osteoblasts is to maintain bone formation and strength, rather than promote osteoclast formation. J Bone Miner Res. 29:1492–1505. 2014. View Article : Google Scholar
12	Fazzalari NL: Bone fracture and bone fracture repair. Osteoporos Int. 22:2003–2006. 2011. View Article : Google Scholar : PubMed/NCBI
13	Franchimont N, Wertz S and Malaise M: Interleukin-6: An osteotropic factor influencing bone formation? Bone. 37:601–606. 2005. View Article : Google Scholar : PubMed/NCBI
14	Holst JJ: The physiology of glucagon-like peptide 1. Physiol Rev. 87:1409–1439. 2007. View Article : Google Scholar : PubMed/NCBI
15	Meier C, Schwartz AV, Egger A and Lecka-Czernik B: Effects of diabetes drugs on the skeleton. Bone. 82:93–100. 2016. View Article : Google Scholar
16	Baggio LL and Drucker DJ: Biology of incretins: GLP-1 and GIP. Gastroenterology. 132:2131–2157. 2007. View Article : Google Scholar : PubMed/NCBI
17	Bollag RJ, Zhong Q, Phillips P, Min L, Zhong L, Cameron R, Mulloy AL, Rasmussen H, Qin F, Ding KH, et al: Osteoblast-derived cells express functional glucose-dependent insulinotropic peptide receptors. Endocrinology. 141:1228–1235. 2000.PubMed/NCBI
18	Bollag RJ, Zhong Q, Ding KH, Phillips P, Zhong L, Qin F, Cranford J, Mulloy AL, Cameron R and Isales CM: Glucose- dependent insulinotropic peptide is an integrative hormone with osteotropic effects. Mol Cell Endocrinol. 177:35–41. 2001. View Article : Google Scholar : PubMed/NCBI
19	Yamada C, Yamada Y, Tsukiyama K, Yamada K, Udagawa N, Takahashi N, Tanaka K, Drucker DJ, Seino Y and Inagaki N: The murine glucagon-like peptide-1 receptor is essential for control of bone resorption. Endocrinology. 149:574–579. 2008. View Article : Google Scholar
20	Sanz C, Vázquez P, Blázquez C, Barrio PA, Alvarez MM and Blázquez E: Signaling and biological effects of glucagon-like peptide 1 on the differentiation of mesenchymal stem cells from human bone marrow. Am J Physiol Endocrinol Metab. 298:E634–E643. 2010. View Article : Google Scholar
21	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
22	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
23	Simpson DA, Feeney S, Boyle C and Stitt AW: Retinal VEGF mRNA measured by SYBR green I fluorescence: A versatile approach to quantitative PCR. Mol Vis. 6:178–183. 2000.PubMed/NCBI
24	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
25	Kato K, Ito H, Hasegawa K, Inaguma Y, Kozawa O and Asano T: Modulation of the stress-induced synthesis of hsp27 and α B-crystallin by cyclic AMP in C6 rat glioma cells. J Neurochem. 66:946–950. 1996. View Article : Google Scholar : PubMed/NCBI
26	Hehlgans T and Pfeffer K: The intriguing biology of the tumour necrosis factor/tumour necrosis factor receptor superfamily: Players, rules and the games. Immunology. 115:1–20. 2005. View Article : Google Scholar : PubMed/NCBI
27	Kobori M, Yang Z, Gong D, Heissmeyer V, Zhu H, Jung YK, Gakidis MA, Rao A, Sekine T, Ikegami F, et al: Wedelolactone suppresses LPS-induced caspase-11 expression by directly inhibiting the IKK complex. Cell Death Differ. 11:123–130. 2004. View Article : Google Scholar
28	Yu Z and Jin T: New insights into the role of cAMP in the production and function of the incretin hormone glucagon-like peptide-1 (GLP-1). Cell Signal. 22:1–8. 2010. View Article : Google Scholar
29	Chijiwa T, Mishima A, Hagiwara M, Sano M, Hayashi K, Inoue T, Naito K, Toshioka T and Hidaka H: Inhibition of forskolin-induced neurite outgrowth and protein phosphorylation by a newly synthesized selective inhibitor of cyclic AMP-dependent protein kinase, N-[2-(p-bromocinnamylamino) ethyl]-5-isoquinolinesulfonamide (H-89), of PC12D pheochromocytoma cells. J Biol Chem. 265:5267–5272. 1990.PubMed/NCBI
30	Nuche-Berenguer B, Portal-Núñez S, Moreno P, González N, Acitores A, López-Herradón A, Esbrit P, Valverde I and Villanueva-Peñacarrillo ML: Presence of a functional receptor for GLP-1 in osteoblastic cells, independent of the cAMP-linked GLP-1 receptor. J Cell Physiol. 225:585–592. 2010. View Article : Google Scholar : PubMed/NCBI
31	Seino Y and Yabe D: Glucose-dependent insulinotropic polypeptide and glucagon-like peptide-1: Incretin actions beyond the pancreas. J Diabetes Investig. 4:108–130. 2013. View Article : Google Scholar : PubMed/NCBI
32	Hoffmann A and Baltimore D: Circuitry of nuclear factor-κB signaling. Immunol Rev. 210:171–186. 2006. View Article : Google Scholar : PubMed/NCBI
33	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