Possible Mechanisms of Di(2-ethylhexyl) Phthalate-Induced MMP-2 and MMP-9 Expression in A7r5 Rat Vascular Smooth Muscle Cells
Abstract
:1. Introduction
2. Results and Discussion
2.1. The Effects of Di(2-ethylhexyl)phthalate (DEHP) on Vascular Smooth Muscle Cells (VSMC) Cell Viability
2.2. The Effects of DEHP on Matrix Metalloproteinase (MMP)-2 and -9 Protein Expression and Their Activities in the VSMC
2.3. Regulatory Pathways of MMP-2 and MMP-9 by DEHP via Phosphorylated p38 Mitogen Activated Protein Kinase (MAPK), Phosphorylated Extracellular Signal Regulated Kinase 1 and 2 (ERK1/2), Phosphorylated Akt, and Nuclear Factor Kappa (NF-κB) (p65)
2.4. The Effects of DEHP on VSMC Migration
3. Experimental Section
3.1. Reagents
3.2. Cell Cultures
3.3. Cell Viability and MMP-2 and MMP-9 Activity Assay
3.4. Western Blotting Analysis of MMP-2, MMP-9, p38 MAPK, ERK1/2, Akt and NF-κB Expression
3.5. Cell Migration Assay
3.6. Statistical Analysis
4. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
- Rudijanto, A. The role of vascular smooth muscle cells on the pathogenesis of atherosclerosis. Acta Med. Indones. 2007, 39, 86–93. [Google Scholar] [PubMed]
- Shah, P.K.; Galis, Z.S. Matrix metalloproteinase hypothesis of plaque rupture: Players keep piling up but questions remain. Circulation 2001, 104, 1878–1880. [Google Scholar] [PubMed]
- Caird, J.; Napoli, C.; Taggart, C.; Farrell, M.; Bouchier-Hayes, D. Matrix metalloproteinases 2 and 9 in human atherosclerotic and non-atherosclerotic cerebral aneurysms. Eur. J. Neurol. 2006, 13, 1098–1105. [Google Scholar] [CrossRef] [PubMed]
- Risinger, G.M., Jr.; Hunt, T.S.; Updike, D.L.; Bullen, E.C.; Howard, E.W. Matrix metalloproteinase-2 expression by vascular smooth muscle cells is mediated by both stimulatory and inhibitory signals in response to growth factors. J. Biol. Chem. 2006, 281, 25915–25925. [Google Scholar] [CrossRef] [PubMed]
- Cho, A.; Reidy, M.A. Matrix metalloproteinase-9 is necessary for the regulation of smooth muscle cell replication and migration after arterial injury. Circ. Res. 2002, 91, 845–851. [Google Scholar] [CrossRef] [PubMed]
- Seo, K.W.; Lee, S.J.; Kim, Y.H.; Bae, J.U.; Park, S.Y.; Bae, S.S.; Kim, C.D. Mechanical stretch increases MMP-2 production in vascular smooth muscle cells via activation of PDGFR-β/Akt signaling pathway. PLoS ONE 2013, 8, e70437. [Google Scholar] [CrossRef] [PubMed]
- Hauser, R.; Calafat, A.M. Phthalates and human health. Occup. Environ. Med. 2005, 62, 806–818. [Google Scholar] [CrossRef] [PubMed]
- Vandenberg, L.N.; Hauser, R.; Marcus, M.; Olea, N.; Welshons, W.V. Human exposure to bisphenol A (BPA). Reprod. Toxicol. 2007, 24, 139–177. [Google Scholar] [CrossRef] [PubMed]
- Latini, G.; del Vecchio, A.; Massaro, M.; Verrotti, A.; de Felice, C. Phthalate exposure and male infertility. Toxicology 2006, 226, 90–98. [Google Scholar] [CrossRef] [PubMed]
- Yao, P.L.; Lin, Y.C.; Richburg, J.H. Mono-(2-ethylhexyl) phthalate-induced disruption of junctional complexes in the seminiferous epithelium of the rodent testis is mediated by MMP2. Biol. Reprod. 2010, 82, 516–527. [Google Scholar] [CrossRef] [PubMed]
- Zhu, H.; Zheng, J.; Xiao, X.; Zheng, S.; Dong, K.; Liu, J.; Wang, Y. Environmental endocrine disruptors promote invasion and metastasis of SK-N-SH human neuroblastoma cells. Oncol. Rep. 2010, 23, 129–139. [Google Scholar] [PubMed]
- Ghosh, J.; Das, J.; Manna, P.; Sil, P.C. Hepatotoxicity of di-(2-ethylhexyl)phthalate is attributed to calcium aggravation, ROS-mediated mitochondrial depolarization, and ERK/NF-κB pathway activation. Free Radic. Biol. Med. 2010, 49, 1779–1791. [Google Scholar] [CrossRef] [PubMed]
- Zhao, J.F.; Hsiao, S.H.; Hsu, M.H.; Pao, K.C.; Kou, Y.R.; Shyue, S.K.; Lee, T.S. Di-(2-ethylhexyl) phthalate accelerates atherosclerosis in apolipoprotein E-deficient mice. Arch. Toxicol. 2014. [Google Scholar] [CrossRef] [PubMed]
- Wang, J.; Liao, Y.; Fan, J.; Ye, T.; Sun, X.; Dong, S. Apigenin inhibits the expression of IL-6, IL-8, and ICAM-1 in DEHP-stimulated human umbilical vein endothelial cells and in vivo. Inflammation 2012, 35, 1466–1476. [Google Scholar] [CrossRef] [PubMed]
- Erkekoglu, P.; Rachidi, W.; Yuzugullu, O.G.; Giray, B.; Favier, A.; Ozturk, M.; Hincal, F. Evaluation of cytotoxicity and oxidative DNA damaging effects of di(2-ethylhexyl)-phthalate (DEHP) and mono(2-ethylhexyl)-phthalate (MEHP) on MA-10 Leydig cells and protection by selenium. Toxicol. Appl. Pharmacol. 2010, 248, 52–62. [Google Scholar] [CrossRef] [PubMed]
- Yao, P.L.; Lin, Y.C.; Richburg, J.H. Mono-(2-ethylhexyl) phthalate (MEHP) promotes invasion and migration of human testicular embryonal carcinoma cells. Biol. Reprod. 2012. [Google Scholar] [CrossRef] [PubMed]
- Blankenberg, S.; Rupprecht, H.J.; Poirier, O.; Bickel, C.; Smieja, M.; Hafner, G.; Meyer, J.; Cambien, F.; Tiret, L. AtheroGene Investigators. Plasma concentrations and genetic variation of matrix metalloproteinase 9 and prognosis of patients with cardiovascular disease. Circulation 2003, 107, 1579–1585. [Google Scholar] [CrossRef] [PubMed]
- Halade, G.V.; Jin, Y.F.; Lindsey, M.L. Matrix metalloproteinase (MMP)-9: A proximal biomarker for cardiac remodeling and a distal biomarker for inflammation. Pharmacol. Ther. 2013, 139, 32–40. [Google Scholar] [CrossRef] [PubMed]
- Johnson, J.L.; George, S.J.; Newby, A.C.; Jackson, C.L. Divergent effects of matrix metalloproteinases 3, 7, 9, and 12 on atherosclerotic plaque stability in mouse brachiocephalic arteries. Proc. Natl. Acad. Sci. USA 2005, 102, 15575–15580. [Google Scholar] [CrossRef] [PubMed]
- Li, Z.; Li, L.; Zielke, H.R.; Cheng, L.; Xiao, R.; Crow, M.T.; Stetler-Stevenson, W.G.; Froehlich, J.; Lakatta, E.G. Increased expression of 72-kd type IV collagenase (MMP-2) in human aortic atherosclerotic lesions. Am. J. Pathol. 1996, 148, 121–128. [Google Scholar] [PubMed]
- Sluijter, J.P.; Pulskens, W.P.; Schoneveld, A.H.; Velema, E.; Strijder, C.F.; Moll, F.; de Vries, J.P.; Verheijen, J.; Hanemaaijer, R.; de Kleijn, D.P.; et al. Matrix metalloproteinase 2 is associated with stable and matrix metalloproteinases 8 and 9 with vulnerable carotid atherosclerotic lesions: A study in human endarterectomy specimen pointing to a role for different extracellular matrix metalloproteinase inducer glycosylation forms. Stroke 2006, 37, 235–239. [Google Scholar] [PubMed]
- Lee, S.O.; Chang, Y.C.; Whang, K.; Kim, C.H.; Lee, I.S. Role of NAD(P)H:quinone oxidoreductase 1 on tumor necrosis factor-α-induced migration of human vascular smooth muscle cells. Cardiovasc. Res. 2007, 76, 331–339. [Google Scholar] [CrossRef] [PubMed]
- Yang, H.L.; Huang, P.J.; Liu, Y.R.; Kumar, K.J.; Hsu, L.S.; Lu, T.L.; Chia, Y.C.; Takajo, T.; Kazunori, A.; Hseu, Y.C. Toona sinensis inhibits LPS-induced inflammation and migration in vascular smooth muscle cells via suppression of reactive oxygen species and NF-κB signaling pathway. Oxidative Med. Cell. Longev. 2014, 2014, 901315. [Google Scholar] [CrossRef] [PubMed]
- Spallarossa, P.; Altieri, P.; Garibaldi, S.; Ghigliotti, G.; Barisione, C.; Manca, V.; Fabbi, P.; Ballestrero, A.; Brunelli, C.; Barsotti, A. Matrix metalloproteinase-2 and -9 are induced differently by doxorubicin in H9c2 cells: The role of MAP kinases and NAD(P)H oxidase. Cardiovasc. Res. 2006, 69, 736–745. [Google Scholar] [CrossRef] [PubMed]
- Cheng, C.Y.; Hsieh, H.L.; Hsiao, L.D.; Yang, C.M. PI3-K/Akt/JNK/NF-κB is essential for MMP-9 expression and outgrowth in human limbal epithelial cells on intact amniotic membrane. Stem Cell Res. 2012, 9, 9–23. [Google Scholar] [CrossRef] [PubMed]
- Yuan, S.; Zhang, J.; Zhang, L.; Huang, L.; Peng, J.; Huang, S.; Chen, S.; Xu, A. The archaic roles of the amphioxus NF-κB/IκB complex in innate immune responses. J. Immunol. 2013, 191, 1220–1230. [Google Scholar] [CrossRef] [PubMed]
- Munshi, H.G.; Wu, Y.; Mukhopadhyay, S.; Ottaviano, A.J.; Sassano, A.; Koblinski, J.E.; Platanias, L.C.; Stack, M.S. Differential regulation of membrane type 1-matrix metalloproteinase activity by ERK 1/2- and p38 MAPK-modulated tissue inhibitor of metalloproteinases 2 expression controls transforming growth factor-beta1-induced pericellular collagenolysis. J. Biol. Chem. 2004, 279, 39042–39050. [Google Scholar] [CrossRef] [PubMed]
- Mukhopadhyay, S.; Munshi, H.G.; Kambhampati, S.; Sassano, A.; Platanias, L.C.; Stack, M.S. Calcium-induced matrix metalloproteinase 9 gene expression is differentially regulated by ERK1/2 and p38 MAPK in oral keratinocytes and oral squamous cell carcinoma. J. Biol. Chem. 2004, 279, 33139–33146. [Google Scholar] [CrossRef] [PubMed]
- Choe, N.; Kwon, J.S.; Kim, J.R.; Eom, G.H.; Kim, Y.; Nam, K.I.; Ahn, Y.; Kee, H.J.; Kook, H. The microRNA miR-132 targets Lrrfip1 to block vascular smooth muscle cell proliferation and neointimal hyperplasia. Atherosclerosis 2013, 229, 348–355. [Google Scholar] [CrossRef] [PubMed]
- Dong, S.; Xiong, W.; Yuan, J.; Li, J.; Liu, J.; Xu, X. MiRNA-146a regulates the maturation and differentiation of vascular smooth muscle cells by targeting NF-κB expression. Mol. Med. Rep. 2013, 8, 407–412. [Google Scholar] [CrossRef] [PubMed]
© 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Shih, M.-F.; Pan, K.-H.; Cherng, J.Y. Possible Mechanisms of Di(2-ethylhexyl) Phthalate-Induced MMP-2 and MMP-9 Expression in A7r5 Rat Vascular Smooth Muscle Cells. Int. J. Mol. Sci. 2015, 16, 28800-28811. https://doi.org/10.3390/ijms161226131
Shih M-F, Pan K-H, Cherng JY. Possible Mechanisms of Di(2-ethylhexyl) Phthalate-Induced MMP-2 and MMP-9 Expression in A7r5 Rat Vascular Smooth Muscle Cells. International Journal of Molecular Sciences. 2015; 16(12):28800-28811. https://doi.org/10.3390/ijms161226131
Chicago/Turabian StyleShih, Mei-Fen, Kuang-Hung Pan, and Jong Yuh Cherng. 2015. "Possible Mechanisms of Di(2-ethylhexyl) Phthalate-Induced MMP-2 and MMP-9 Expression in A7r5 Rat Vascular Smooth Muscle Cells" International Journal of Molecular Sciences 16, no. 12: 28800-28811. https://doi.org/10.3390/ijms161226131