IL-1α induces apoptosis and inhibits the osteoblast differentiation of MC3T3-E1 cells through the JNK and p38 MAPK pathways

Guo,Chun; Yang,Xu-Guang; Wang,Fei; Ma,Xu-Yuan

doi:10.3892/ijmm.2016.2606

IL-1α induces apoptosis and inhibits the osteoblast differentiation of MC3T3-E1 cells through the JNK and p38 MAPK pathways

Authors:
- Chun Guo
- Xu-Guang Yang
- Fei Wang
- Xu-Yuan Ma
View Affiliations

Affiliations: Department of Medicine, Luohe Medical College, Luohe, Henan 462002, P.R. China, Huaihe Hospital, Henan University, Kaifeng, Henan 475000, P.R. China
Published online on: May 25, 2016 https://doi.org/10.3892/ijmm.2016.2606
Pages: 319-327

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

Interleukin (IL)-1 is a proinflammatory cytokine that plays important roles in inflammation and host responses to infection. The present study aimed to evaluate the effects of IL-1α on the apoptosis and differentiation of osteoblasts, and to elucidate the mechanism responsible for these effects in the osteoblast‑like cell line MC3T3-E1. The MC3T3-E1 cells were non-treated or treated with IL-1α. Following treatment, cell viability, alkaline phosphatase (ALP) activity and caspase-3 activity were evaluated. The expression of osteoblast-specific genes as well as Bax, Bcl-2 and caspase-3 were determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The protein levels of Bax, Bcl-2, caspase-3 and the phosphorylation of mitogen-activated protein kinases (MAPKs, also known as MAP kinases) were evaluated using western blot analysis. The MAPK signaling pathway was blocked by pre-treatment with MAPK inhibitors SB203580, PD98059 and SP600125. IL-1α treatment induced a significant decrease in cell viability and ALP activity in the MC3T3-E1 cells. IL-1α also significantly decreased the mRNA expression and protein levels of osteoblast-related genes in the MC3T3-E1 cells. On the other hand, IL-1α significantly upregulated the mRNA expression and protein levels of Bax and caspase-3 as well as caspase-3 activity, whereas Bcl-2 expression was decreased in the MC3T3-E1 cells. Furthermore, IL-1α activated the apoptotic signaling pathway by increasing the phosphorylation of c-Jun N-terminal kinase (JNK) and p38-MAPK, whereas it inhibited the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2). Moreover, pre-treatment with MAPK inhibitors attenuated the phosphorylation of JNK, p38 and Bax expression enhanced by IL-1α. However, MAPK inhibitors markedly increased the protein expression of osteoblast-related genes and Bcl-xL in the MC3T3-E1 cells downregulated by IL-1α. Taken together, these findings suggest that IL-1α induces the apoptosis of osteoblasts and inhibits osteoblast differentiation by activating the JNK and the p38 MAPK pathways.

View Figures

View References

1	Roodman GD: Advances in bone biology: the osteoclast. Endocr Rev. 17:308–332. 1996.PubMed/NCBI
2	Nair SP, Meghji S, Wilson M, Reddi K, White P and Henderson B: Bacterially induced bone destruction: mechanisms and misconceptions. Infect Immun. 64:2371–2380. 1996.PubMed/NCBI
3	Dinarello CA: Biologic basis for interleukin-1 in disease. Blood. 87:2095–2147. 1996.PubMed/NCBI
4	Nakae S, Asano M, Horai R and Iwakura Y: Interleukin-1 beta, but not interleukin-1 alpha, is required for T-cell-dependent antibody production. Immunology. 104:402–409. 2001. View Article : Google Scholar
5	Garlanda C, Dinarello CA and Mantovani A: The interleukin-1 family: back to the future. Immunity. 39:1003–1018. 2013. View Article : Google Scholar : PubMed/NCBI
6	Stashenko P, Dewhirst FE, Rooney ML, Desjardins LA and Heeley JD: Interleukin-1 beta is a potent inhibitor of bone formation in vitro. J Bone Miner Res. 2:559–565. 1987. View Article : Google Scholar : PubMed/NCBI
7	Ohmori Y, Hanazawa S, Amano S, Hirose K, Kumegawa M and Kitano S: Effects of recombinant human interleukin I alpha and interleukin 1 beta on cell growth and alkaline phosphatase of the mouse osteoblastic cell line MC3T3-E1. Biochim Biophys Acta. 970:22–30. 1988. View Article : Google Scholar : PubMed/NCBI
8	Lacey DL, Grosso LE, Moser SA, Erdmann J, Tan HL, Pacifici R and Villareal DT: IL-1-induced murine osteoblast IL-6 production is mediated by the type 1 IL-1 receptor and is increased by 1,25 dihydroxyvitamin D3. J Clin Invest. 91:1731–1742. 1993. View Article : Google Scholar : PubMed/NCBI
9	Lee YM, Fujikado N, Manaka H, Yasuda H and Iwakura Y: IL-1 plays an important role in the bone metabolism under physiological conditions. Int Immunol. 22:805–816. 2010. View Article : Google Scholar : PubMed/NCBI
10	Gowen M, Wood DD, Ihrie EJ, McGuire MK and Russell RG: An interleukin 1-like factor stimulates bone resorption in vitro. Nature. 306:378–380. 1983. View Article : Google Scholar : PubMed/NCBI
11	Lorenzo JA, Sousa SL, Alander C, Raisz LG and Dinarello CA: Comparison of the bone-resorbing activity in the supernatants from phytohemagglutinin-stimulated human peripheral blood mononuclear cells with that of cytokines through the use of an antiserum to interleukin 1. Endocrinology. 121:1164–1170. 1987. View Article : Google Scholar : PubMed/NCBI
12	Nakamura I and Jimi E: Regulation of osteoclast differentiation and function by interleukin-1. Vitam Horm. 74:357–370. 2006. View Article : Google Scholar : PubMed/NCBI
13	Matsuguchi T, Chiba N, Bandow K, Kakimoto K, Masuda A and Ohnishi T: JNK activity is essential for Atf4 expression and late-stage osteoblast differentiation. J Bone Miner Res. 24:398–410. 2009. View Article : Google Scholar
14	Guo C, Yuan L, Wang JG, Wang F, Yang XK, Zhang FH, Song JL, Ma XY, Cheng Q and Song GH: Lipopolysaccharide (LPS) induces the apoptosis and inhibits osteoblast differentiation through JNK pathway in MC3T3-E1 cells. Inflammation. 37:621–631. 2014. View Article : Google Scholar
15	Guo C, Wang SL, Xu ST, Wang JG and Song GH: SP600125 reduces lipopolysaccharide-induced apoptosis and restores the early-stage differentiation of osteoblasts inhibited by LPS through the MAPK pathway in MC3T3-E1 cells. Int J Mol Med. 35:1427–1434. 2015.PubMed/NCBI
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
17	Komori T and Kishimoto T: Cbfa1 in bone development. Curr Opin Genet Dev. 8:494–499. 1998. View Article : Google Scholar : PubMed/NCBI
18	Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K, Shimizu Y, Bronson RT, Gao YH, Inada M, et al: Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell. 89:755–764. 1997. View Article : Google Scholar : PubMed/NCBI
19	Nakashima K, Zhou X, Kunkel G, Zhang Z, Deng JM, Behringer RR and de Crombrugghe B: The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation. Cell. 108:17–29. 2002. View Article : Google Scholar : PubMed/NCBI
20	Fu H, Doll B, McNelis T and Hollinger JO: Osteoblast differentiation in vitro and in vivo promoted by Osterix. J Biomed Mater Res A. 83:770–778. 2007. View Article : Google Scholar : PubMed/NCBI
21	Kim HK, Cho SG, Kim JH, Doan TK, Hu QS, Ulhaq R, Song EK and Yoon TR: Mevinolin enhances osteogenic genes (ALP, type I collagen and osteocalcin), CD44, CD47 and CD51 expression during osteogenic differentiation. Life Sci. 84:290–295. 2009. View Article : Google Scholar : PubMed/NCBI
22	Pearson G, Robinson F, Beers Gibson T, Xu BE, Karandikar M, Berman K and Cobb MH: Mitogen-activated protein (MAP) kinase pathways: regulation and physiological functions. Endocr Rev. 22:153–183. 2001.PubMed/NCBI
23	Kyriakis JM and Avruch J: Mammalian MAPK signal transduction pathways activated by stress and inflammation: a 10-year update. Physiol Rev. 92:689–737. 2012. View Article : Google Scholar : PubMed/NCBI
24	Xia Z, Dickens M, Raingeaud J, Davis RJ and Greenberg ME: Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science. 270:1326–1331. 1995. View Article : Google Scholar : PubMed/NCBI
25	Tang C, Liang J, Qian J, Jin L, Du M, Li M and Li D: Opposing role of JNK-p38 kinase and ERK1/2 in hydrogen peroxide-induced oxidative damage of human trophoblast-like JEG-3 cells. Int J Clin Exp Pathol. 7:959–968. 2014.PubMed/NCBI
26	Fister S, Günthert AR, Aicher B, Paulini KW, Emons G and Gründker C: GnRH-II antagonists induce apoptosis in human endometrial, ovarian, and breast cancer cells via activation of stress-induced MAPKs p38 and JNK and proapoptotic protein Bax. Cancer Res. 69:6473–6481. 2009. View Article : Google Scholar : PubMed/NCBI