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Involvement of calcium-sensing receptor in oxLDL-induced MMP-2 production in vascular smooth muscle cells via PI3K/Akt pathway

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Abstract

Matrix metalloproteinase-2 (MMP-2) is constitutively expressed in vascular smooth muscle cells (VSMCs) and up-regulated in atherosclerotic lesion by various stimuli, such as oxidized low-density lipoprotein (oxLDL). Calcium-sensing receptor (CaSR) is also expressed in VSMCs, but it remains unclear whether CaSR is associated with overproduction of MMP-2 in VSMCs. In this study, the expression of MMP-2 was detected by real-time PCR and Western blot analysis, and the gelatinolytic activity of MMP-2 was measured using gelatin zymography. Our results showed that oxLDL enhanced MMP-2 expression and activity in rat aortic VSMCs in a time- and dose-dependent manner. In addition, CaSR expression was up-regulated by oxLDL. Manipulating CaSR function in these cells by NPS2390 (an antagonist of CaSR) or GdCl3 (an agonist of CaSR) affected the oxLDL-induced MMP-2 production. In VSMCs, oxLDL stimulated the rapid activation of phosphatidylinositol 3-kinase (PI3K)/Akt signal pathway, as determined by Western blot analysis. Phosphorylation of Akt and MMP-2 production stimulated by oxLDL were attenuated by LY294002 (a specific inhibitor of PI3K). Activation of Akt was suppressed by NPS2390 but enhanced by GdCl3. In contrast, oxLDL had no stimulatory effect on the phosphorylation of JNK, and pretreatment with SP600125 (an inhibitor of JNK) produced no significant effect on oxLDL-induced MMP-2 production. These results suggest that CaSR mediates oxLDL-induced MMP-2 production in VSMCs via PI3K/Akt signal pathway.

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References

  1. Auge N, Maupas-Schwalm F, Elbaz M, Thiers JC, Waysbort A, Itohara S, Krell HW, Salvayre R, Negre-Salvayre A (2004) Role for matrix metalloproteinase-2 in oxidized low-density lipoprotein-induced activation of the sphingomyelin/ceramide pathway and smooth muscle cell proliferation. Circulation 110:571–578

    Article  PubMed  CAS  Google Scholar 

  2. Aoyagi M, Yamamoto M, Azuma H, Nagashima G, Niimi Y, Tamaki M, Hirakawa K, Yamamoto K (1998) Immunolocalization of matrix metalloproteinases in rabbit carotid arteries after balloon denudation. Histochem Cell Biol 109:97–102

    Article  PubMed  CAS  Google Scholar 

  3. Li Z, Li L, Zielke HR, Cheng L, Xiao R, Crow MT, Stetler-Stevenson WG, Froehlich J, Lakatta EG (1996) Increased expression of 72-kd type IV collagenase (MMP-2) in human aortic atherosclerotic lesions. Am J Pathol 148:121–128

    PubMed  CAS  Google Scholar 

  4. Ikeda U, Shimada K (2003) Matrix metalloproteinases and coronary artery diseases. Clin Cardiol 26:55–59

    Article  PubMed  Google Scholar 

  5. Brown EM, Gamba G, Riccardi D, Lombardi M, Butters R, Kifor O, Sun A, Hediger MA, Lytton J, Hebert SC (1993) Cloning and characterization of an extracellular Ca2+-sensing receptor from bovine parathyroid. Nature 366:575–580

    Article  PubMed  CAS  Google Scholar 

  6. Brown EM (2000) The extracellular Ca2+-sensing receptor: central mediator of systemic calcium homeostasis. Annu Rev Nutr 20:507–533

    Article  PubMed  CAS  Google Scholar 

  7. Xi YH, Li HZ, Zhang WH, Wang LN, Zhang L, Lin Y, Bai SZ, Li HX, Wu LY, Wang R, Xu CQ (2010) The functional expression of calcium-sensing receptor in the differentiated THP-1 cells. Mol Cell Biochem 342:233–240

    Article  PubMed  CAS  Google Scholar 

  8. Tu CL, Chang W, Xie Z, Bikle DD (2008) Inactivation of the calcium sensing receptor inhibits E-cadherin-mediated cell–cell adhesion and calcium-induced differentiation in human epidermal keratinocytes. J Biol Chem 283:3519–3528

    Article  PubMed  CAS  Google Scholar 

  9. Molostvov G, Fletcher S, Bland R, Zehnder D (2008) Extracellular calcium-sensing receptor mediated signalling is involved in human vascular smooth muscle cell proliferation and apoptosis. Cell Physiol Biochem 22:413–422

    Article  PubMed  CAS  Google Scholar 

  10. Cifuentes M, Rojas CV (2008) Antilipolytic effect of calcium-sensing receptor in human adipocytes. Mol Cell Biochem 319:17–21

    Article  PubMed  CAS  Google Scholar 

  11. Wang R, Xu C, Zhao W, Zhang J, Cao K, Yang B, Wu L (2003) Calcium and polyamine regulated calcium-sensing receptors in cardiac tissues. Eur J Biochem 270:2680–2688

    Article  PubMed  CAS  Google Scholar 

  12. Tfelt-Hansen J, Hansen JL, Smajilovic S, Terwilliger EF, Haunso S, Sheikh SPN (2006) Calcium receptor is functionally expressed in rat neonatal ventricular cardiomyocytes. Am J Physiol Heart Circ Physiol 290:H1165–H1171

    Article  PubMed  CAS  Google Scholar 

  13. Molostvov G, Bland R, Zehnder D (2009) Expression and role of the calcium-sensing receptor in the blood vessel wall. Curr Pharm Biotechnol 10:282–288

    Article  PubMed  CAS  Google Scholar 

  14. Yamaguchi T, Kifor O, Chattopadhyay N, Bai M, Brown EM (1998) Extracellular calcium (Ca 2+o )-sensing receptor in a mouse monocyte-macrophage cell line (J774): potential mediator of the actions of Ca 2+o on the function of J774 cells. J Bone Miner Res 13:1390–1397

    Article  PubMed  CAS  Google Scholar 

  15. Guo J, Li HZ, Zhang WH, Wang LC, Wang LN, Zhang L, Li GW, Li HX, Yang BF, Wu L, Wang R, Xu CQ (2010) Increased expression of calcium-sensing receptors induced by ox-LDL amplifies apoptosis of cardiomyocytes during simulated ischaemia-reperfusion. Clin Exp Pharmacol Physiol 37:e128–e135

    Article  PubMed  CAS  Google Scholar 

  16. Witztum JL, Steinberg D (1991) Role of oxidized low density lipoprotein in atherogenesis. J Clin Investig 88:1785–1792

    Article  PubMed  CAS  Google Scholar 

  17. Rudijanto A (2007) The role of vascular smooth muscle cells on the pathogenesis of atherosclerosis. Acta Med Indones 39:86–93

    PubMed  Google Scholar 

  18. Robbesyn F, Auge N, Vindis C, Cantero AV, Barbaras R, Negre-Salvayre A, Salvayre R (2005) High-density lipoproteins prevent the oxidized low-density lipoprotein-induced endothelial growth factor receptor activation and subsequent matrix metalloproteinase-2 upregulation. Arterioscler Thromb Vasc Biol 25:1206–1212

    Article  PubMed  CAS  Google Scholar 

  19. Pauly RR, Bilato C, Cheng L, Monticone R, Crow MT (1997) Vascular smooth muscle cell cultures. Methods Cell Biol 52:133–154

    Article  PubMed  CAS  Google Scholar 

  20. Lee SJ, Seo KW, Yun MR, Bae SS, Lee WS, Hong KW, Kim CD (2008) 4-Hydroxynonenal enhances MMP-2 production in vascular smooth muscle cells via mitochondrial ROS-mediated activation of the Akt/NF-kappaB signaling pathways. Free Radic Biol Med 45:1487–1492

    Article  PubMed  CAS  Google Scholar 

  21. Wang LN, Wang C, Lin Y, Xi YH, Zhang WH, Zhao YJ, Li HZ, Tian Y, Lv YJ, Yang BF, Xu CQ (2008) Involvement of calcium-sensing receptor in cardiac hypertrophy-induced by angiotensinII through calcineurin pathway in cultured neonatal rat cardiomyocytes. Biochem Biophys Res Commun 369:584–589

    Article  PubMed  CAS  Google Scholar 

  22. Raffetto JD, Khalil RA (2008) Matrix metalloproteinases and their inhibitors in vascular remodeling and vascular disease. Biochem Pharmacol 75:346–359

    Article  PubMed  CAS  Google Scholar 

  23. Beaudeux JL, Giral P, Bruckert E, Foglietti MJ, Chapman MJ (2004) Matrix metalloproteinases, inflammation and atherosclerosis: therapeutic perspectives. Clin Chem Lab Med 42:121–131

    Article  PubMed  CAS  Google Scholar 

  24. Risinger GM Jr, Hunt TS, Updike DL, Bullen EC, Howard EW (2006) 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 281:25915–25925

    Article  PubMed  CAS  Google Scholar 

  25. Lu Q, Hong W (2009) Bcl2 enhances c-Myc-mediated MMP-2 expression of vascular smooth muscle cells. Cell Signal 21:1054–1059

    Article  PubMed  CAS  Google Scholar 

  26. Molostvov G, James S, Fletcher S, Bennett J, Lehnert H, Bland R, Zehnder D (2007) Extracellular calcium-sensing receptor is functionally expressed in human artery. Am J Physiol Ren Physiol 293:F946–F955

    Article  CAS  Google Scholar 

  27. Lee SJ, Bae SS, Kim KH, Lee WS, Rhim BY, Hong KW, Kim CD (2007) High glucose enhances MMP-2 production in adventitial fibroblasts via Akt1-dependent NF-kappaB pathway. FEBS Lett 581:4189–4194

    Article  PubMed  CAS  Google Scholar 

  28. Cellurale C, Sabio G, Kennedy NJ, Das M, Barlow M, Sandy P, Jacks T, Davis RJ (2011) Requirement of c-Jun NH(2)-terminal kinase for Ras-initiated tumor formation. Mol Cell Biol 31:1565–1576

    Article  PubMed  CAS  Google Scholar 

  29. Xiao LW, Yang M, Dong J, Xie H, Sui GL, He YL, Lei JX, Liao EY, Yuan X (2011) Stretch-inducible expression of connective tissue growth factor (CTGF) in human osteoblasts-like cells is mediated by PI3K-JNK pathway. Cell Physiol Biochem 28:297–304

    Article  PubMed  CAS  Google Scholar 

  30. Takabe W, Li R, Ai L, Yu F, Berliner J, Hsiai TK (2010) Oxidized low-density lipoprotein-activated c-Jun NH2-terminal kinase regulates manganese superoxide dismutase ubiquitination: implication for mitochondrial redox status and apoptosis. Arterioscler Thromb Vasc Biol 30:436–441

    Article  PubMed  CAS  Google Scholar 

  31. Boudot C, Saidak Z, Boulanouar AK, Petit L, Gouilleux F, Massy Z, Brazier M, Mentaverri R, Kamel S (2010) Implication of the calcium sensing receptor and the phosphoinositide 3-kinase/Akt pathway in the extracellular calcium-mediated migration of RAW 264.7 osteoclast precursor cells. Bone 46:1416–1423

    Article  PubMed  CAS  Google Scholar 

  32. Thyberg J (1998) Phenotypic modulation of smooth muscle cells during formation of neointimal thickenings following vascular injury. Histol Histopathol 13:871–891

    PubMed  CAS  Google Scholar 

  33. Hultgardh-Nilsson A, Lovdahl C, Blomgren K, Kallin B, Thyberg J (1997) Expression of phenotype- and proliferation-related genes in rat aortic smooth muscle cells in primary culture. Cardiovasc Res 34:418–430

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This study is supported by the National Natural Science Foundation of China (30871012, 81070123, 81170178) and the National Basic Research Program of China (973 Program, 2007CB512000).

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Authors and Affiliations

  1. Department of Pathophysiology, Harbin Medical University, Harbin, 150086, China

    Hong-Xia Li, Fan-Juan Kong, Shu-Zhi Bai, Wen He, Wen-Jing Xing, Yu-Hui Xi, Guang-Wei Li, Jin Guo, Hong-Zhu Li, Ling-Yun Wu, Rui Wang, Ye Tian & Chang-Qing Xu

  2. Bio-Pharmaceutical Key Laboratory of Heilongjiang Province, Harbin, 150086, China

    Chang-Qing Xu

  3. Department of Biology, Lakehead University, Thunder Bay, ON, P7B 5E1, Canada

    Ling-Yun Wu, Rui Wang & Guang-Dong Yang

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  1. Hong-Xia Li
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Correspondence to Chang-Qing Xu.

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Hong-Xia Li and Fan-Juan Kong are considered as co-first authors.

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Li, HX., Kong, FJ., Bai, SZ. et al. Involvement of calcium-sensing receptor in oxLDL-induced MMP-2 production in vascular smooth muscle cells via PI3K/Akt pathway. Mol Cell Biochem 362, 115–122 (2012). https://doi.org/10.1007/s11010-011-1133-6

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  • DOI: https://doi.org/10.1007/s11010-011-1133-6

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