Molecular Medicine

, Volume 21, Issue 1, pp 616–625 | Cite as

Angiotensin II Downregulates MicroRNA-145 to Regulate Kruppel-like Factor 4 and Myocardin Expression in Human Coronary Arterial Smooth Muscle Cells under High Glucose Conditions

  • Kou-Gi Shyu
  • Wen-Ping Cheng
  • Bao-Wei Wang
Research Article


MicroRNA (miR)-145 is the most abundant miR in vascular smooth muscle cells (VSMCs). However, the effect of hyperglycemia on the regulation of miR-145 is unknown. We hypothesized that the hyperglycemic condition activates a proinflammatory response that mediates the expression of miR-145 in VSMCs. We investigated whether miR-145 serves as a critical regulator to regulate the downstream proliferation factors (including Kruppel-like factor 4 (Klf4) and myocardin) in VSMCs under hyperglycemic conditions. Human coronary artery smooth muscle cells (HCASMCs) were cultured under high glucose conditions. Sustained high glucose at 25 mmol/L significantly decreased the expression of miR-145 in HCASMCs. High glucose significantly increased angiotensin II (Ang II) secretion from HCASMCs and Ang II suppressed miR-145 expression in HCASMCs. Ang II repression of miR145 expression resulted in increased Klf4 and decreased myocardin expression under conditions of high glucose. Overexpression of miR-145 significantly decreased Klf4 and increased myocardin expression and inhibited HCASMC proliferation and migration induced by a high glucose state. Balloon injury of the carotid artery in diabetic rats was performed to investigate miR-145, Klf and myocardin expression. The expression of miR-145 was maximally increased at 7 d after carotid injury and gradually declined thereafter. Overexpression of miR-145 and treatment with valsartan reversed Klf4 and myocardin protein expression induced by balloon injury and improved vascular injury. In conclusion, our study reveals that Ang II downregulates miR-145 to regulate Klf4 and myocardin expression in HCASMCs under high glucose conditions. Ang II plays a critical role in the regulation of miR-145 under hyperglycemic conditions.



This study was supported by grants from Ministry of Science and Technology, Taiwan and Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan.

Supplementary material

10020_2015_2101616_MOESM1_ESM.pdf (2.9 mb)
Supplementary material, approximately 2.88 MB.


  1. 1.
    Paneni F, Beckman JA, Creager MA, Cosentino F. (2013) Diabetes and vascular disease: pathophysiology, clinical consequences, and medical therapy: part 1. Eur. Heart J. 34:2436–46.CrossRefGoogle Scholar
  2. 2.
    Shntikumarn S, Caporali A, Emanueli C. (2012) Role of microRNAs in diabetes and its cardiovascular complications. Cardiovasc. Res. 93:583–93.CrossRefGoogle Scholar
  3. 3.
    Zampetaki A, Mayr M. (2012) MicroRNAs in vascular and metabolic disease. Circ. Res. 110:508–22.CrossRefGoogle Scholar
  4. 4.
    Boettger T, et al. (2009) Acquisition of the contractile phenotype by murine arterial smooth muscle cells depends on the Mir143/145 gene cluster. J. Clin. Invest. 119:2634–47.CrossRefGoogle Scholar
  5. 5.
    Cordes KR, et al. (2009) miR-145 and miR-143 regulate smooth muscle cell fate and plasticity. Nature. 460:705–10.PubMedPubMedCentralGoogle Scholar
  6. 6.
    Cheng Y, et al. (2009) MicroRNA-145, a novel smooth muscle cell phenotypic marker and modulator, controls vascular neointimal lesion formation. Circ. Res. 105:158–66.CrossRefGoogle Scholar
  7. 7.
    Xin M, et al. (2009) MicroRNAs miR-143 and miR-145 modulate cytoskelet al dynamics and responsiveness of smooth muscle cells to injury. Genes. Dev. 23:2166–78.CrossRefGoogle Scholar
  8. 8.
    Wang Z, et al. (2004) Myocardin and ternary complex factors compete for SRF to control smooth muscle gene expression. Nature. 428:185–9.CrossRefGoogle Scholar
  9. 9.
    Garvey SM, Sinden DS, Schoppee Bortz PD, Wamhoff BR. (2010) Cyclosporine up-regulates Kruppel-like factor-4 (KLF4) in vascular smooth muscle cells and drives phenotypic modulation in vivo. J. Pharmacol. Exp. Ther. 333:34–42.CrossRefGoogle Scholar
  10. 10.
    Zanella MT, Kohlmann O Jr., Ribeiro AB. (2001) Treatment of obesity, hypertension and diabetes syndrome. Hypertension. 38:705–8.CrossRefGoogle Scholar
  11. 11.
    Shyu KG, Wang BW, Wu GJ, Lin CM, Chang H. (2013) Mechanical stretch via transforming growth factor-β 1 activates microRNA208a to regulate endoglin expression in cultured rat cardiac myoblasts. Eur. J. Heart Fail. 15:36–45.CrossRefGoogle Scholar
  12. 12.
    Cheng WP, Hung HF, Wang BW, Shyu KG. (2008) The molecular regulation of GADD153 in apoptosis of cultured vascular smooth muscle cells by cyclic mechanical stretch. Cardiovasc. Res. 77:551–9.CrossRefGoogle Scholar
  13. 13.
    Wang BW, Chang H, Lin S, Kuan P, Shyu KG. (2003) Induction of matrix met alloproteinase-14 and -2 by cyclical mechanical stretch is mediated by tumor necrosis factor-α in cultured human umbilical vein endothelial cells. Cardiovasc. Res. 59:460–9.CrossRefGoogle Scholar
  14. 14.
    Chang H, et al. (2003) GL-331 inhibits HIF-1α expression in a lung cancer model. Biochem. Biophys. Res. Commun. 302:95–100.CrossRefGoogle Scholar
  15. 15.
    Shyu KG, Wang BW, Kuan P, Chang H. (2008) RNA Interference for discoidin domain receptor 2 attenuates neointimal formation in balloon injured rat carotid artery. Arterioscler. Thromb. Vasc. Biol. 28:1447–53.CrossRefGoogle Scholar
  16. 16.
    Committee for the Update of the Guide for the Care and Use of Laboratory Animals, Institute for Laboratory Animal Research, Division on Earth and Life Studies, National Research Council of the National Academies. (2011) Guide for the Care and Use of Laboratory Animals. 8th edition. Washington (DC): National Academic Press.Google Scholar
  17. 17.
    Hu B, et al. (2014) Mechanical stretch suppresses microRNA-145 expression by activating extracellular signal-regulated kinase1/2 and upregulating angiotensin-converting enzyme to alter vascular smooth muscle cell phenotype. PLoS One. 9:e96338.CrossRefGoogle Scholar
  18. 18.
    Davies-Dusenbery BN, et al. (2011) Downregulation of Kruppel-like factor-4 (KLF4) by microRNA-143/145 is critical for modulation of vascular smooth muscle cell phenotype by transforming growth factor-β and bone morphogenetic protein 4. J. Biol. Chem. 286:28097–110.CrossRefGoogle Scholar
  19. 19.
    Rangrez AY, Massy ZA, Meuth VML, Metzinger L. (2011) miR-143 and miR-145: Molecular keys to switch the phenotype of vascular smooth muscle cells. Circ. Cardiovasc. Genet. 4:197–205.CrossRefGoogle Scholar
  20. 20.
    Shu HJ, Wen JK, Miao SB, Liu Y, Zheng B. (2012) KLF5 and hhLIM cooperatively promote proliferation of vascular smooth muscle cells. Mol. Cell Biochem. 367:185–94.CrossRefGoogle Scholar
  21. 21.
    Bartel DP. (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 116:281–97.CrossRefGoogle Scholar
  22. 22.
    Natarajan R, Putta S, Kato M. (2012) MicroRNAs and diabetic complication. J. Cardiovasc. Trans. Res. 5:413–22.CrossRefGoogle Scholar
  23. 23.
    Small EM, Olson EN. (2007) Pervasive roles of microRNAs in cardiovascular biology. Nature. 469:36–342.Google Scholar
  24. 24.
    Ji R, et al. (2007) MicroRNA expression signature and antisense-mediated depletion reveal an essential role of microRNA in vascular neointimal lesion formation. Circ. Res. 100:1579–88.CrossRefGoogle Scholar
  25. 25.
    Rzucidlo EM, Martin KA, Powell RJ. (2007) Regulation of vascular smooth muscle cell differentiation. J. Vasc. Surg. 45:A25–32.CrossRefGoogle Scholar
  26. 26.
    Hutcheson R, et al. (2013) MicroRNA-145 restores contractile vascular smooth muscle phenotype and coronary collateral growth in the metabolic syndrome. Arterioscler. Thromb. Vasc. Biol. 33:727–36.CrossRefGoogle Scholar
  27. 27.
    Natarajan R, Scott S, Bai W, Yerneni KKV, Nadler J. (1999) Angiotensin II signaling in vascular smooth muscle cells under high glucose conditions. Hypertension. 33:378–84.CrossRefGoogle Scholar
  28. 28.
    Sodhi CP, Kanwar YS, Sahai A. (2003) Hypoxia and high glucose upregulate AT1 receptor expression and potentiate ANG II-induced proliferation in VSM cells. Am. J. Physiol. Heart Circ. Physiol. 284:H846–52.CrossRefGoogle Scholar
  29. 29.
    Xue, et al. (2013) H2S inhibits hyperglycemia-induced intrarenal renin-angiotensin system activation via attenuation of reactive oxygen species generation. PLoS One. 8:e74336.CrossRefGoogle Scholar
  30. 30.
    Wen et al. (2014) miRNA-145 is involved in the development of resistin-induced insulin resistance in HepG2 cells. Biochem. Biophys. Res. Commun. 445:517–23.CrossRefGoogle Scholar
  31. 31.
    Norta GD, et al. (2012) MicroRNA 143-145 deficiency impairs vascular function. Int. J. Immunol. Pharmacol. 25:467–84.Google Scholar
  32. 32.
    Lovren F, et al. (2012) MicroRNA-145 targeted therapy reduces atherosclerosis Circulation. 126(11 Suppl 1):S81–90.CrossRefGoogle Scholar

Copyright information

© The Author(s) 2015

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (

Authors and Affiliations

  1. 1.Division of CardiologyShin Kong Wu Ho-Su Memorial HospitalTaipeiTaiwan
  2. 2.Graduate Institute of Clinical MedicineTaipei Medical UniversityTaipeiTaiwan
  3. 3.Department of Medical Education and ResearchShin Kong Wu Ho-Su Memorial HospitalTaipeiTaiwan

Personalised recommendations