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Journal of Nephrology

, Volume 32, Issue 1, pp 101–110 | Cite as

Activation of the mTORC1 pathway by inflammation contributes to vascular calcification in patients with end-stage renal disease

  • Jing Liu
  • Wei Zhu
  • Chun Ming Jiang
  • Yuan Feng
  • Yang Yang Xia
  • Qing Yan Zhang
  • Miao ZhangEmail author
Original Article
  • 120 Downloads

Abstract

Background

Chronic inflammation plays an important role in the progression of vascular calcification (VC). This study was designed to explore the effects and underlying mechanisms of inflammation on VC in the radial arteries of patients with end-stage renal disease (ESRD) with arteriovenostomy.

Methods

Forty-eight ESRD patients were divided into control (n = 25) and inflammation groups (n = 23) according to plasma C-reactive protein (CRP) level. Surgically removed tissues from the radial arteries of patients receiving arteriovenostomy were used in this study. Alizarin Red S staining was used to examine calcium deposition. The expression of inflammation markers, bone structure-associated proteins and mammalian target of rapamycin complex1 (mTORC1) pathway-related proteins was assessed by immunohistochemical staining.

Results

The expression of tumor necrosis factor-α (TNF-α) and monocyte chemotactic protein-1 (MCP-1) was increased in the radial arteries of the inflammation group. Additionally, Alizarin Red S staining revealed a marked increase in calcium deposition in the inflammation group compared to controls. Further analysis by immunohistochemical staining demonstrated that the deposition was correlated with the increased expression of bone-associated proteins such as bone morphogenetic proteins-2 (BMP-2) and osteocalcin and collagen I, which suggested that inflammation induces osteogenic differentiation in vascular tissues and that osteogenic cells are the main cellular components involved in VC. Interestingly, there was a parallel increase in the expression of phosphorylated mTOR (p-mTOR) and pribosomal protein S6 kinase 1 (p-S6K1) in the inflammation group. Furthermore, mTORC1 pathway-related proteins were significantly associated with the enhanced expression of bone formation biomarkers.

Conclusions

Inflammation contributed to VC in the radial arteries of ESRD patients via the induction of osteogenic differentiation in vessel walls, which could be regulated by the activation of the mTORC1 pathway.

Keywords

Inflammation Vascular calcification mTORC1 pathway End-stage renal disease 

Notes

Author contributions

MZ and JL conceived and designed the experiments; JL performed the experiments; JL and WZ analyzed the data; CJ, YF, YX and QZ contributed reagents/materials/analysis tools; JL wrote the paper.

Funding

This study was supported by grants from the National Natural Science Foundation of China (No.81500585), and Nanjing Medical Science and Technique Development Foundation (No.QRX17120).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

All studies were approved by the Ethical Committee of Nanjing Drum Tower Hospital. Additional informed consent was obtained from all individual participants for whom identifying information is included in this article.

References

  1. 1.
    Foley RN, Murray AM, Li S, Herzog CA, McBean AM, Eggers PW, Collins AJ (2005) Chronic kidney disease and the risk for cardiovascular disease, renal replacement, and death in the United States Medicare population, 1998 to 1999. J Am Soc Nephrol 16(2):489–495.  https://doi.org/10.1681/ASN.2004030203 CrossRefGoogle Scholar
  2. 2.
    Stenvinkel P, Carrero JJ, Axelsson J, Lindholm B, Heimbürger O, Massy Z (2008) Emerging biomarkers for evaluating cardiovascular risk in the chronic kidney disease patient: how do new pieces t into the uremic puzzle? Clin J Am Soc Nephrol 3(2):505–521.  https://doi.org/10.2215/CJN.03670807 CrossRefGoogle Scholar
  3. 3.
    Kaysen GA (2001) The microinflammatory state in uremia: causes and potential consequences. J Am Soc Nephrol 12(7):1549–1557Google Scholar
  4. 4.
    Chen NC, Hsu CY, Chen CL (2017) The strategy to prevent and regress the vascular calcification in dialysis patients. Biomed Res Int 2017:9035193.  https://doi.org/10.1155/2017/9035193
  5. 5.
    Yamada S, Tokumoto M, Tatsumoto N, Taniguchi M, Noguchi H, Nakano T, Masutani K, Ooboshi H, Tsuruya K, Kitazono T (2014) Phosphate overload directly induces systemic inflammation and malnutrition as well as vascular calcification in uremia. Am J Physiol Renal Physiol 306(12):1418–1428.  https://doi.org/10.1152/ajprenal.00633.2013 CrossRefGoogle Scholar
  6. 6.
    Yoshihara F (2016) Systemic inflammation is a key factor for mortality risk stratification in chronic kidney disease patients with coronary artery calcification. Circ J 80(7):1537–1538.  https://doi.org/10.1253/circj.CJ-16-0506 CrossRefGoogle Scholar
  7. 7.
    Evrard S, Delanaye P, Kamel S, Cristol JP, Cavalier E (2015) Vascular calcification: from pathophysiology to biomarkers. Clin Chim Acta 438:401–414.  https://doi.org/10.1016/j.cca.2014.08.034 CrossRefGoogle Scholar
  8. 8.
    Lau WL, Ix JH (2013) Clinical detection, risk factors, and cardiovascular consequences of medial arterial calci cation: a pattern of vascular injury associated with aberrant mineral metabolism. Semin Nephrol 33(2):93–105.  https://doi.org/10.1016/j.semnephrol.2012.12.011 CrossRefGoogle Scholar
  9. 9.
    Steitz SA, Speer MY, Curinga G, Yang HY, Haynes P, Aebersold R, Schinke T, Karsenty G, Giachelli CM (2001) Smooth muscle cell phenotypic transition associated with calcification: upregulation of Cbfa1 and downregulation of smooth muscle lineage markers. Circ Res 89(12):1147–1154CrossRefGoogle Scholar
  10. 10.
    Neven E, Dauwe S, De Broe ME, D’Haese PC, Persy V (2007) Endochondral bone formation is involved in media calcification in rats and in men. Kidney Int 72(5):574–581.  https://doi.org/10.1038/sj.ki.5002353 CrossRefGoogle Scholar
  11. 11.
    Towler DA (2013) Molecular and cellular aspects of calcific aortic valve disease. Circ Res 113(2):198–208.  https://doi.org/10.1161/CIRCRESAHA.CrossRefGoogle Scholar
  12. 12.
    Barreto FC, Barreto DV, Moyses RM, Neves CL, Jorgetti V, Draibe SA, Canziani ME, Carvalho AB (2006) Osteoporosis in hemodial-ysis patients revisited by bone histomorphometry: a new insight into an old problem. Kidney Int 69(10):1852–1857CrossRefGoogle Scholar
  13. 13.
    Cai T, Sun D, Duan Y, Wen P, Dai C, Yang J, He W (2016) WNT/β-catenin signaling promotes VSMCs to osteogenic transdifferentiation and calcification through directly modulating Runx2 gene expression. Exp Cell Res 345(2):206–217.  https://doi.org/10.1016/j.yexcr.2016.06.007 CrossRefGoogle Scholar
  14. 14.
    Khosla S (2011) The bone and beyond: a shift in calcium. Nat Med 17(4):430–431.  https://doi.org/10.1038/nm0411-430 CrossRefGoogle Scholar
  15. 15.
    Hruska KA, Mathew S, Lund RJ, Memon I, Saab G (2009) The pathogenesis of vascular calcification in the chronic kidney disease mineral bone disorder: the links between bone and the vasculature. Semin Nephrol 29(2):156–165.  https://doi.org/10.1016/j.semnephrol CrossRefGoogle Scholar
  16. 16.
    Alesutan I, Voelkl J, Feger M, Kratschmar DV, Castor T, Mia S, Sacherer M, Viereck R, Borst O, Leibrock C, Gawaz M, Kuro-O M, Pilz S, Tomaschitz A, Odermatt A, Pieske B, Wagner CA, Lang F (2017) Involvement of vascular aldosterone synthase in phosphate-induced osteogenic transformation of vascular smooth muscle cells. Sci Rep 7(1):2059.  https://doi.org/10.1038/s41598-017-01882-2 CrossRefGoogle Scholar
  17. 17.
    Jewell JL, Guan KL (2013) Nutrient signaling to mTOR and cell growth. Trends Biochem Sci 38(5):233–242.  https://doi.org/10.1016/j.tibs.2013.01.004 CrossRefGoogle Scholar
  18. 18.
    Zhao Y, Zhao MM, Cai Y, Zheng MF, Sun WL, Zhang SY, Kong W, Gu J, Wang X, Xu MJ (2015) Mammalian target of rapamycin signaling inhibition ameliorates vascular calcification via Klotho upregulation. Kidney Int 88(4):711–721.  https://doi.org/10.1038/ki.2015.160 CrossRefGoogle Scholar
  19. 19.
    Vervloet MG, Adema AY, Larsson TE, Massy ZA (2014) The role of klotho on vascular calcification and endothelial function in chronic kidney disease. Semin Nephrol 34(6):578–585.  https://doi.org/10.1016/j.semnephrol.2014.09.003 CrossRefGoogle Scholar
  20. 20.
    Yeh LC, Ma X, Ford JJ, Adamo ML, Lee JC (2013) Rapamycin inhibits BMP-7-induced osteogenic and lipogenic marker expressions in fetal rat calvarial cells. J Cell Biochem 114(8):1760–1771.  https://doi.org/10.1002/jcb.24519 CrossRefGoogle Scholar
  21. 21.
    Ma KL, Liu J, Wang CX, Ni J, Zhang Y, Wu Y, Lv LL, Ruan XZ, Liu BC (2013) Activation of mTOR modulates SREBP-2 to induce foam cell formation through increased retinoblastoma protein phosphorylation. Cardiovasc Res 100(3):450–460.  https://doi.org/10.1093/cvr/cvt203 CrossRefGoogle Scholar
  22. 22.
    Liu J, Ma KL, Zhang Y, Wu Y, Hu ZB, Lv LL, Tang RN, Liu H, Ruan XZ, Liu BC (2015) Activation of mTORC1 disrupted LDL receptor pathway: a potential new mechanism for the progression of non-alcoholic fatty liver disease. Int J Biochem Cell Biol 61:8–19.  https://doi.org/10.1016/j.biocel.2015.01.011 CrossRefGoogle Scholar
  23. 23.
    Jung HH, Kim SW, Han H (2006) Inflammation, mineral metabolism and progressive coronary artery calcification in patients on haemodialysis. Nephrol Dial Transplant 21(7):1915–1920.  https://doi.org/10.1093/ndt/gfl118 CrossRefGoogle Scholar
  24. 24.
    Liu J, Ma KL, Gao M, Wang CX, Ni J, Zhang Y, Zhang XL, Liu H, Wang YL, Liu BC (2012) Inflammation disrupts the LDL receptor pathway and accelerates the progression of vascular calcification in ESRD patients. PLoS One 7(10):47217.  https://doi.org/10.1371/journal.pone.0047217 CrossRefGoogle Scholar
  25. 25.
    Din ASharafE, Salem UA, Abdulazim MM DO (2016) Vascular calcification: when should we interfere in chronic kidney disease patients and how? World J Nephrol 5(5):398–417.  https://doi.org/10.5527/wjn.v5.i5.398 CrossRefGoogle Scholar
  26. 26.
    Hwang IC, Park HE, Kim HL, Kim HM, Park JB, Yoon YE, Lee SP, Kim HK, Cho GY, Sohn DW, Kim YJ (2016) Systemic in ammation is associated with coronary artery calcifica-tion and all-cause mortality in chronic kidney disease. Circ J 80(7):1644–1652.  https://doi.org/10.1253/circj.CJ-15-1224 CrossRefGoogle Scholar
  27. 27.
    Martínez-Moreno JM1, Muñoz-Castañeda JR, Herencia C, Oca AM, Estepa JC, Canalejo R, Rodríguez-Ortiz ME, Perez-Martinez P, Aguilera-Tejero E, Canalejo A, Rodríguez M, Almadén Y (2012) In vascular smooth muscle cells paricalcitol prevents phosphate-induced Wnt/β-catenin activation. Am J Physiol Renal Physiol 303(8):1136–1144.  https://doi.org/10.1152/ajprenal.00684.2011 CrossRefGoogle Scholar
  28. 28.
    Viaene L, Behets GJ, Heye S, Claes K, Monbaliu D, Pirenne J, D’Haese PC, Evenepoel P (2016) Inflammation and the bone-vascular axis in end-stage renal disease. Osteoporos Int 27(2):489–497.  https://doi.org/10.1007/s00198-015-3233-8.CrossRefGoogle Scholar
  29. 29.
    Cheng L, Zhang L, Yang J, Hao L (2017) Activation of peroxisome proliferatoractivated receptor γ inhibits vascular calcification by upregulating Klotho. Exp Ther Med 13(2):467–474.  https://doi.org/10.3892/etm.2016.3996 CrossRefGoogle Scholar
  30. 30.
    Montezano AC, Touyz RM (2014) Mammalian target of rapamycin: a novel pathway in vascular calcification. Can J Cardiol 30(5):482–484.  https://doi.org/10.1016/j.cjca.2014.03.001 CrossRefGoogle Scholar
  31. 31.
    Hu Y, Lou J, Mao YY, Lai TW, Liu LY, Zhu C, Zhang C, Liu J, Li YY, Zhang F, Li W, Ying SM, Chen ZH, Shen HH (2016) Activation of MTOR in pulmonary epithelium promotes LPS-induced acute lung injury. Autophagy 12(12):2286–2299.  https://doi.org/10.1080/15548627.2016.1230584 docCrossRefGoogle Scholar
  32. 32.
    Xiao Z, Peng J, Gan N, Arafat A, Yin F (2016) Interleukin-1β plays a pivotal role via the PI3K/Akt/mTOR signaling pathway in the chronicity of mesial temporal lobe epilepsy. Neuroimmunomodulation 23(5–6):332–344.  https://doi.org/10.1159/000460254 CrossRefGoogle Scholar
  33. 33.
    Xian L, Wu X, Pang L, Lou M, Rosen CJ, Qiu T, Crane J, Frassica F, Zhang L, Rodriguez JP, Jia X, Yakar S, Xuan S, Efstratiadis A, Wan M, Cao X (2012) Matrix IGF-1 maintains bone mass by activation of mTOR in mesenchymal stem cells. Nat Med 18(7):1095–1101.  https://doi.org/10.1038/nm.2793 CrossRefGoogle Scholar
  34. 34.
    Zhan JK, Wang YJ, Wang Y, Wang S, Tan P, Huang W, Liu YS (2014) The mammalian target of rapamycin signalling pathway is involved in osteoblastic differentiation of vascular smooth muscle cells. Can J Cardiol 30(5):568–575.  https://doi.org/10.1016/j.cjca.2013.11.005 CrossRefGoogle Scholar

Copyright information

© Italian Society of Nephrology 2018

Authors and Affiliations

  • Jing Liu
    • 1
  • Wei Zhu
    • 1
  • Chun Ming Jiang
    • 1
  • Yuan Feng
    • 1
  • Yang Yang Xia
    • 1
  • Qing Yan Zhang
    • 1
  • Miao Zhang
    • 1
    Email author
  1. 1.Institute of Nephrology, Affiliated Drum Tower HospitalMedical School of Nanjing UniversityNanjingChina

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