Zinc Deficiency Promoted Fibrosis via ROS and TIMP/MMPs in the Myocardium of Mice

  • Jing-wen Cao
  • Shi-yu Duan
  • Hong-xin Zhang
  • Yu Chen
  • Mengyao GuoEmail author


Zinc (Zn) is an important trace element in the body that has antioxidant effects. It has been proven that Zn deficiency can cause oxidative stress. The purpose of the present study was to clarify the effect and mechanism of Zn deficiency on myocardial fibrosis. Mice were fed with different Zn levels dietary for 9 weeks: Zn-normal group (ZnN, 34 mg Zn/kg), Zn-deficient group (ZnD, 2 mg Zn/kg), and Zn-adequate group (ZnA, 100 mg Zn/kg). We found that the Zn-deficient diet reduced the Zn concentration in myocardial tissue. Moreover, the TUNEL results demonstrated that cardiomyocytes in the ZnD group died in large numbers. Furthermore, ROS levels were significantly increased, and metallothionein (MT) expression levels decreased in the ZnD group. The results of Sirius Red staining indicated an increase in collagen in the ZnD group. Moreover, the ELISA results showed that collagen I, III, and IV and fibronectin (FN) were increased. In addition, the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinase (TIMPs) was detected by RT-qPCR. The results showed that the expression of TIMP-1 in the ZnD group was increased, while MMPs were decreased. Immunohistochemical results showed an increase in the content of α-smooth muscle actin (α-SMA), while H&E staining showed an increase in interstitial width and a decrease in the number of cardiac cells. All data suggest that Zn deficiency enhances the oxidative stress response of myocardial tissue and eventually triggers myocardial fibrosis.


Zn deficiency Myocardial fibrosis Oxygen radicals (ROS) Matrix metalloproteinases (MMPs) Extracellular matrix (ECM) 


Compliance with Ethical Standards

All animals were obtained from Huazhong Agricultural University Laboratory Animal Research Center (Wuhan, China), according to the animal welfare standards of use, and approved by the Ethical Committee on Animal Research at Huazhong Agricultural University (HZAUMO-2015-12).


  1. 1.
    Writing C, Smith SC Jr, Collins A, Ferrari R, Holmes DR Jr, Logstrup S et al (2012) Our time: a call to save preventable death from cardiovascular disease (heart disease and stroke). Glob Heart 7:297–305CrossRefGoogle Scholar
  2. 2.
    Gyongyosi M, Winkler J, Ramos I, Do QT, Firat H, McDonald K et al (2017) Myocardial fibrosis: biomedical research from bench to bedside. Eur J Heart Fail 19:177–191CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Weber KT, Sun Y, Bhattacharya SK, Ahokas RA, Gerling IC (2013) Myofibroblast-mediated mechanisms of pathological remodelling of the heart. Nat Rev Cardiol 10:15–26CrossRefPubMedGoogle Scholar
  4. 4.
    Cohn JN, Ferrari R, Sharpe N (2000) Cardiac remodeling—concepts and clinical implications: a consensus paper from an international forum on cardiac remodeling. J Am Coll Cardiol 35:569–582CrossRefPubMedGoogle Scholar
  5. 5.
    Gonzalez A, Ravassa S, Beaumont J, Lopez B, Diez J (2011) New targets to treat the structural remodeling of the myocardium. J Am Coll Cardiol 58:1833–1843CrossRefPubMedGoogle Scholar
  6. 6.
    Zhao S, Wu H, Xia W, Chen X, Zhu S, Zhang S, Shao Y, Ma W, Yang D, Zhang J (2014) Periostin expression is upregulated and associated with myocardial fibrosis in human failing hearts. J Cardiol 63:373–378CrossRefPubMedGoogle Scholar
  7. 7.
    Li AH, Liu PP, Villarreal FJ, Garcia RA (2014) Dynamic changes in myocardial matrix and relevance to disease: translational perspectives. Circ Res 114:916–927CrossRefPubMedGoogle Scholar
  8. 8.
    Xiang FL, Fang M, Yutzey KE (2017) Loss of beta-catenin in resident cardiac fibroblasts attenuates fibrosis induced by pressure overload in mice. Nat Commun 8:712CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Zebrowski DC, Engel FB (2013) The cardiomyocyte cell cycle in hypertrophy, tissue homeostasis, and regeneration. Rev Physiol Biochem Pharmacol 165:67–96CrossRefPubMedGoogle Scholar
  10. 10.
    Kuhn B, del Monte F, Hajjar RJ, Chang YS, Lebeche D, Arab S et al (2007) Periostin induces proliferation of differentiated cardiomyocytes and promotes cardiac repair. Nat Med 13:962–969CrossRefPubMedGoogle Scholar
  11. 11.
    Shamhart PE, Meszaros JG (2010) Non-fibrillar collagens: key mediators of post-infarction cardiac remodeling? J Mol Cell Cardiol 48:530–537CrossRefPubMedGoogle Scholar
  12. 12.
    Nagase H, Visse R, Murphy G (2006) Structure and function of matrix metalloproteinases and TIMPs. Cardiovasc Res 69:562–573CrossRefPubMedGoogle Scholar
  13. 13.
    Visse R, Nagase H (2003) Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function, and biochemistry. Circ Res 92:827–839CrossRefPubMedGoogle Scholar
  14. 14.
    Scrimgeour AG, Carrigan CT, Condlin ML, Urso ML, van den Berg RM, van Helden HPM, Montain SJ, Joosen MJA (2018) Dietary zinc modulates matrix metalloproteinases in traumatic brain injury. J Neurotrauma 35:2495–2506CrossRefPubMedGoogle Scholar
  15. 15.
    Yoshida A, Kanamori H, Naruse G, Minatoguchi S, Iwasa M, Yamada Y, Mikami A, Kawasaki M, Nishigaki K, Minatoguchi S (2019) (Pro)renin receptor blockade ameliorates heart failure caused by chronic kidney disease. J Card Fail 25:286–300CrossRefPubMedGoogle Scholar
  16. 16.
    Jarosz M, Olbert M, Wyszogrodzka G, Mlyniec K, Librowski T (2017) Antioxidant and anti-inflammatory effects of zinc. Zinc-dependent NF-kappaB signaling. Inflammopharmacology 25:11–24CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Vasak M, Meloni G (2011) Chemistry and biology of mammalian metallothioneins. J Biol Inorganic Chem : JBIC : a publication of the Society of Biological Inorganic Chemistry 16:1067–1078CrossRefGoogle Scholar
  18. 18.
    Ji SG, Weiss JH (2018) Zn2+-induced disruption of neuronal mitochondrial function: synergism with Ca2+, critical dependence upon cytosolic Zn2+ buffering, and contributions to neuronal injury. Exp Neurol 302:181–195CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Castro L, Freeman B (2001) Reactive oxygen species in human health and disease. Nutrition 17:161–165CrossRefPubMedGoogle Scholar
  20. 20.
    Lorenzen JM, Schauerte C, Hubner A, Kolling M, Martino F, Scherf K et al (2015) Osteopontin is indispensible for AP1-mediated angiotensin II-related miR-21 transcription during cardiac fibrosis. Eur Heart J 36:2184–2196CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Xiang FL, Guo MZ, Yutzey KE (2016) Overexpression of Tbx20 in adult cardiomyocytes promotes proliferation and improves cardiac function after myocardial infarction. Circulation 133:1081–1092CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Tomat AL, Costa MD, Arranz CT (2011) Zinc restriction during different periods of life: influence in renal and cardiovascular diseases. Nutrition 27:392–398CrossRefPubMedGoogle Scholar
  23. 23.
    Maret W (2000) The function of zinc metallothionein: a link between cellular zinc and redox state. J Nutr 130:1455s–1458sCrossRefPubMedGoogle Scholar
  24. 24.
    Oliveira HCF, Cosso RG, Alberici LC, Maciel EN, Salerno AG, Dorighello GG et al (2004) Oxidative stress in atherosclerosis-prone mouse is due to low antioxidant capacity of mitochondria. FASEB J 18:278Google Scholar
  25. 25.
    Sato M, Kondoh M (2002) Recent studies on metallothionein: protection against toxicity of heavy metals and oxygen free radicals. Tohoku J Exp Med 196:9–22CrossRefPubMedGoogle Scholar
  26. 26.
    Yang LF, Ma JP, Tan Y, Zheng QJ, Dong ML, Guo W et al (2019) Cardiac-specific overexpression of metallothionein attenuates L-NAME-induced myocardial contractile anomalies and apoptosis. J Cell Mol Med 23:4640–4652PubMedPubMedCentralGoogle Scholar
  27. 27.
    Laity JH, Andrews GK (2007) Understanding the mechanisms of zinc-sensing by metal-response element binding transcription factor-1 (MTF-1). Arch Biochem Biophys 463:201–210CrossRefPubMedGoogle Scholar
  28. 28.
    Prasad AS (2008) Zinc in human health: effect of zinc on immune cells. Mol Med 14:353–357CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Frangogiannis NG (2019) Cardiac fibrosis: cell biological mechanisms, molecular pathways and therapeutic opportunities. Mol Asp Med 65:70–99CrossRefGoogle Scholar
  30. 30.
    Li L, Zhao Q, Kong W (2018) Extracellular matrix remodeling and cardiac fibrosis. Matrix Biol 68-69:490–506CrossRefPubMedGoogle Scholar
  31. 31.
    Malemud CJ (2006) Matrix metalloproteinases (MMPs) in health and disease: an overview. Front Biosci-Landmrk 11:1696–1701CrossRefGoogle Scholar
  32. 32.
    Peterson JT, Li H, Dillon L, Bryant JW (2000) Evolution of matrix metalloprotease and tissue inhibitor expression during heart failure progression in the infarcted rat. Cardiovasc Res 46:307–315CrossRefPubMedGoogle Scholar
  33. 33.
    Lovelock JD, Baker AH, Gao F, Dong JF, Bergeron AL, McPheat W, Sivasubramanian N, Mann DL (2005) Heterogeneous effects of tissue inhibitors of matrix metalloproteinases on cardiac fibroblasts. Am J Physiol-Heart C 288:H461–H468CrossRefGoogle Scholar
  34. 34.
    Karamanos NK, Theocharis AD, Neill T, Iozzo RV (2019) Matrix modeling and remodeling: a biological interplay regulating tissue homeostasis and diseases. Matrix Biol 75-76:1–11CrossRefPubMedGoogle Scholar
  35. 35.
    Barbolina MV, Stack MS (2008) Membrane type 1-matrix metalloproteinase: substrate diversity in pericellular proteolysis. Semin Cell Dev Biol 19:24–33CrossRefPubMedGoogle Scholar
  36. 36.
    Moore L, Fan D, Basu R, Kandalam V, Kassiri Z (2012) Tissue inhibitor of metalloproteinases (TIMPs) in heart failure. Heart Fail Rev 17:693–706CrossRefPubMedGoogle Scholar
  37. 37.
    Sivasubramanian N, Coker ML, Kurrelmeyer KM, MacLellan WR, DeMayo FJ, Spinale FG et al (2001) Left ventricular remodeling in transgenic mice with cardiac restricted overexpression of tumor necrosis factor. Circulation 104:826–831CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Clinical Veterinary Medicine, College of Veterinary MedicineHuazhong Agricultural UniversityWuhanPeople’s Republic of China

Personalised recommendations