Myocardial infarction is a frequent complication of cardiovascular disease leading to high morbidity and mortality worldwide. Elevated C-reactive protein (CRP) levels after myocardial infarction are associated with heart failure and poor prognosis. Cardiomyocyte microvesicles (CMV) are released during hypoxic conditions and can act as mediators of intercellular communication. MicroRNA (miRNA) are short non-coding RNA which can alter cellular mRNA-translation. Microvesicles (MV) have been shown to contain distinct patterns of miRNA from their parent cells which can affect protein expression in target cells. We hypothesized that miRNA containing CMV mediate hepatic CRP expression after cardiomyocyte hypoxia. H9c2-cells were cultured and murine cardiomyocytes were isolated from whole murine hearts. H9c2- and murine cardiomyocytes were exposed to hypoxic conditions using a hypoxia chamber. Microvesicles were isolated by differential centrifugation and analysed by flow cytometry. Next-generation-sequencing was performed to determine the miRNA-expression profile in H9c2 CMV compared to their parent cells. Microvesicles were incubated with a co-culture model of the liver consisting of THP-1 macrophages and HepG2 cells. IL-6 and CRP expression in the co-culture was assessed by qPCR and ELISA. CMV contain a distinct pattern of miRNA compared to their parent cells including many inflammation-related miRNA. CMV induced IL-6 expression in THP-1 macrophages alone and CRP expression in the hepatic co-culture model. MV from hypoxic cardiomyocytes can mediate CRP expression in a hepatic co-culture model. Further studies will have to show whether these effects are reproducible in-vivo.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Availability of data and material
All data is available from the authors upon reasonable request.
Benjamin EJ, Muntner P, Alonso A, Bittencourt MS, Callaway CW, Carson AP, Chamberlain AM, Chang AR, Cheng S, Das SR, Delling FN, Djousse L, Elkind MSV, Ferguson JF, Fornage M, Jordan LC, Khan SS, Kissela BM, Knutson KL, Kwan TW, Lackland DT, Lewis TT, Lichtman JH, Longenecker CT, Loop MS, Lutsey PL, Martin SS, Matsushita K, Moran AE, Mussolino ME, O'Flaherty M, Pandey A, Perak AM, Rosamond WD, Roth GA, Sampson UKA, Satou GM, Schroeder EB, Shah SH, Spartano NL, Stokes A, Tirschwell DL, Tsao CW, Turakhia MP, VanWagner LB, Wilkins JT, Wong SS, Virani SS (2019) Heart disease and stroke statistics—2019 update: a report from the American Heart Association. Circulation 139(10):e56–e528. https://doi.org/10.1161/cir.0000000000000659
Granger CB, Kochar A (2018) Understanding and targeting inflammation in acute myocardial infarction. An elusive goal. J Am Coll Cardiol 72(2):199–201. https://doi.org/10.1016/j.jacc.2018.05.006
Bursi F, Weston SA, Killian JM, Gabriel SE, Jacobsen SJ, Roger VL (2007) C-reactive protein and heart failure after myocardial infarction in the community. Am J Med 120(7):616–622. https://doi.org/10.1016/j.amjmed.2006.07.039
Suleiman M, Khatib R, Agmon Y, Mahamid R, Boulos M, Kapeliovich M, Levy Y, Beyar R, Markiewicz W, Hammerman H, Aronson D (2006) Early inflammation and risk of long-term development of heart failure and mortality in survivors of acute myocardial infarction predictive role of C-reactive protein. J Am Coll Cardiol 47(5):962–968. https://doi.org/10.1016/j.jacc.2005.10.055
Volanakis JE (2001) Human C-reactive protein: expression, structure, and function. Mol Immunol 38(2–3):189–197. https://doi.org/10.1016/s0161-5890(01)00042-6
Thompson D, Pepys MB, Wood SP (1999) The physiological structure of human C-reactive protein and its complex with phosphocholine. Structure 7(2):169–177. https://doi.org/10.1016/s0969-2126(99)80023-9
Black S, Kushner I, Samols D (2004) C-reactive protein. J Biol Chem 279(47):48487–48490. https://doi.org/10.1074/jbc.R400025200
Sturk A, Hack CE, Aarden LA, Brouwer M, Koster RR, Sanders GT (1992) Interleukin-6 release and the acute-phase reaction in patients with acute myocardial infarction: a pilot study. J Lab Clin Med 119(5):574–579
Marcoux G, Duchez AC, Cloutier N, Provost P, Nigrovic PA, Boilard E (2016) Revealing the diversity of extracellular vesicles using high-dimensional flow cytometry analyses. Sci Rep 6:35928. https://doi.org/10.1038/srep35928
Yu H, Wang Z (2019) Cardiomyocyte-derived exosomes: biological functions and potential therapeutic implications. Front Physiol 10:1049–1049. https://doi.org/10.3389/fphys.2019.01049
Diehl P, Fricke A, Sander L, Stamm J, Bassler N, Htun N, Ziemann M, Helbing T, El-Osta A, Jowett JB, Peter K (2012) Microparticles: major transport vehicles for distinct microRNAs in circulation. Cardiovasc Res 93(4):633–644. https://doi.org/10.1093/cvr/cvs007
Loyer X, Zlatanova I, Devue C, Yin M, Howangyin K-Y, Klaihmon P, Guerin CL, Kheloufi M, Vilar J, Zannis K, Fleischmann BK, Hwang DW, Park J, Lee H, Menasché P, Silvestre J-S, Boulanger CM (2018) Intra-cardiac release of extracellular vesicles shapes inflammation following myocardial infarction. Circ Res 123(1):100–106. https://doi.org/10.1161/CIRCRESAHA.117.311326
Yang J, Yu X, Xue F, Li Y, Liu W, Zhang S (2018) Exosomes derived from cardiomyocytes promote cardiac fibrosis via myocyte-fibroblast cross-talk. Am J Transl Res 10(12):4350–4366
Yu X, Deng L, Wang D, Li N, Chen X, Cheng X, Yuan J, Gao X, Liao M, Wang M, Liao Y (2012) Mechanism of TNF-alpha autocrine effects in hypoxic cardiomyocytes: initiated by hypoxia inducible factor 1alpha, presented by exosomes. J Mol Cell Cardiol 53(6):848–857. https://doi.org/10.1016/j.yjmcc.2012.10.002
Waldenström A, Gennebäck N, Hellman U, Ronquist G (2012) Cardiomyocyte microvesicles contain DNA/RNA and convey biological messages to target cells. PLoS ONE 7(4):e34653. https://doi.org/10.1371/journal.pone.0034653
Chaturvedi P, Kalani A, Medina I, Familtseva A, Tyagi SC (2015) Cardiosome mediated regulation of MMP9 in diabetic heart: role of mir29b and mir455 in exercise. J Cell Mol Med 19(9):2153–2161. https://doi.org/10.1111/jcmm.12589
Giricz Z, Varga ZV, Baranyai T, Sipos P, Pálóczi K, Kittel Á, Buzás EI, Ferdinandy P (2014) Cardioprotection by remote ischemic preconditioning of the rat heart is mediated by extracellular vesicles. J Mol Cell Cardiol 68:75–78. https://doi.org/10.1016/j.yjmcc.2014.01.004
Zordoky BN, El-Kadi AO (2007) H9c2 cell line is a valuable in vitro model to study the drug metabolizing enzymes in the heart. J Pharmacol Toxicol Methods 56(3):317–322. https://doi.org/10.1016/j.vascn.2007.06.001
O'Connell TD, Rodrigo MC, Simpson PC (2007) Isolation and culture of adult mouse cardiac myocytes. Methods Mol Biol 357:271–296. https://doi.org/10.1385/1-59745-214-9:271
Riccardi C, Nicoletti I (2006) Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat Protoc 1(3):1458–1461. https://doi.org/10.1038/nprot.2006.238
Camps C, Buffa FM, Colella S, Moore J, Sotiriou C, Sheldon H, Harris AL, Gleadle JM, Ragoussis J (2008) hsa-miR-210 Is induced by hypoxia and is an independent prognostic factor in breast cancer. Clin Cancer Res 14(5):1340–1348. https://doi.org/10.1158/1078-0432.ccr-07-1755
Yu X, Deng L, Wang D, Li N, Chen X, Cheng X, Yuan J, Gao X, Liao M, Wang M, Liao Y (2012) Mechanism of TNF-α autocrine effects in hypoxic cardiomyocytes: Initiated by hypoxia inducible factor 1α, presented by exosomes. J Mol Cell Cardiol 53(6):848–857. https://doi.org/10.1016/j.yjmcc.2012.10.002
Gupta S, Knowlton AA (2007) HSP60 trafficking in adult cardiac myocytes: role of the exosomal pathway. Am J Physiol Heart Circ Physiol 292(6):H3052–H3056. https://doi.org/10.1152/ajpheart.01355.2006
Valadi H, Ekstrom K, Bossios A, Sjostrand M, Lee JJ, Lotvall JO (2007) Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol 9(6):654–659. https://doi.org/10.1038/ncb1596
Zhang J, Chiodini R, Badr A, Zhang G (2011) The impact of next-generation sequencing on genomics. J Genet Genomics 38(3):95–109. https://doi.org/10.1016/j.jgg.2011.02.003
Li C, Li J, Xue K, Zhang J, Wang C, Zhang Q, Chen X, Gao C, Yu X, Sun L (2019) MicroRNA-143-3p promotes human cardiac fibrosis via targeting sprouty3 after myocardial infarction. J Mol Cell Cardiol 129:281–292. https://doi.org/10.1016/j.yjmcc.2019.03.005
Parahuleva MS, Euler G, Mardini A, Parviz B, Schieffer B, Schulz R, Aslam M (2017) Identification of microRNAs as potential cellular monocytic biomarkers in the early phase of myocardial infarction: a pilot study. Sci Rep 7(1):15974. https://doi.org/10.1038/s41598-017-16263-y
Das S, Kohr M, Dunkerly-Eyring B, Lee DI, Bedja D, Kent OA, Leung AK, Henao-Mejia J, Flavell RA, Steenbergen C (2017) Divergent effects of miR-181 family members on myocardial function through protective cytosolic and detrimental mitochondrial microRNA targets. J Am Heart Assoc. https://doi.org/10.1161/jaha.116.004694
Yuan J, Chen H, Ge D, Xu Y, Xu H, Yang Y, Gu M, Zhou Y, Zhu J, Ge T, Chen Q, Gao Y, Wang Y, Li X, Zhao Y (2017) Mir-21 promotes cardiac fibrosis after myocardial infarction via targeting Smad7. Cell Physiol Biochem 42(6):2207–2219. https://doi.org/10.1159/000479995
Liu X, Dong Y, Chen S, Zhang G, Zhang M, Gong Y, Li X (2015) Circulating MicroRNA-146a and MicroRNA-21 predict left ventricular remodeling after ST-elevation myocardial infarction. Cardiology 132(4):233–241. https://doi.org/10.1159/000437090
Pfeiffer D, Roßmanith E, Lang I, Falkenhagen D (2017) miR-146a, miR-146b, and miR-155 increase expression of IL-6 and IL-8 and support HSP10 in an In vitro sepsis model. PLoS ONE 12(6):e0179850–e0179850. https://doi.org/10.1371/journal.pone.0179850
Tukov FF, Maddox JF, Amacher DE, Bobrowski WF, Roth RA, Ganey PE (2006) Modeling inflammation-drug interactions in vitro: a rat Kupffer cell-hepatocyte coculture system. Toxicol In Vitro 20(8):1488–1499. https://doi.org/10.1016/j.tiv.2006.04.005
Matak P, Chaston TB, Chung B, Srai SK, McKie AT, Sharp PA (2009) Activated macrophages induce hepcidin expression in HuH7 hepatoma cells. Haematologica 94(6):773–780. https://doi.org/10.3324/haematol.2008.003400
Couch Y, Evans MC, Gardiner C, Sargent I, Losey P, Lambertsen KL, Anthony DC (2014) Brain-derived microvesicles confer sickness behaviours by switching on the acute phase response in the liver. J Neuroimmunol 275(1):57. https://doi.org/10.1016/j.jneuroim.2014.08.150
Neri T, Armani C, Pegoli A, Cordazzo C, Carmazzi Y, Brunelleschi S, Bardelli C, Breschi MC, Paggiaro P, Celi A (2011) Role of NF-kappaB and PPAR-gamma in lung inflammation induced by monocyte-derived microparticles. Eur Respir J 37(6):1494–1502. https://doi.org/10.1183/09031936.00023310
Fink K, Moebes M, Vetter C, Bourgeois N, Schmid B, Bode C, Helbing T, Busch H-J (2015) Selenium prevents microparticle-induced endothelial inflammation in patients after cardiopulmonary resuscitation. Crit Care 19(1):58–58. https://doi.org/10.1186/s13054-015-0774-3
Ikeda U, Ohkawa F, Seino Y, Yamamoto K, Hidaka Y, Kasahara T, Kawai T, Shimada K (1992) Serum interleukin 6 levels become elevated in acute myocardial infarction. J Mol Cell Cardiol 24(6):579–584. https://doi.org/10.1016/0022-2828(92)91042-4
Shu J, Ren N, Du JB, Zhang M, Cong HL, Huang TG (2007) Increased levels of interleukin-6 and matrix metalloproteinase-9 are of cardiac origin in acute coronary syndrome. Scand Cardiovasc J 41(3):149–154. https://doi.org/10.1080/14017430601164263
Suleiman M, Aronson D, Reisner SA, Kapeliovich MR, Markiewicz W, Levy Y, Hammerman H (2003) Admission C-reactive protein levels and 30-day mortality in patients with acute myocardial infarction. Am J Med 115(9):695–701. https://doi.org/10.1016/j.amjmed.2003.06.008
Thiele JR, Habersberger J, Braig D, Schmidt Y, Goerendt K, Maurer V, Bannasch H, Scheichl A, Woollard KJ, von Dobschutz E, Kolodgie F, Virmani R, Stark GB, Peter K, Eisenhardt SU (2014) Dissociation of pentameric to monomeric C-reactive protein localizes and aggravates inflammation: in vivo proof of a powerful proinflammatory mechanism and a new anti-inflammatory strategy. Circulation 130(1):35–50. https://doi.org/10.1161/circulationaha.113.007124
Habersberger J, Strang F, Scheichl A, Htun N, Bassler N, Merivirta RM, Diehl P, Krippner G, Meikle P, Eisenhardt SU, Meredith I, Peter K (2012) Circulating microparticles generate and transport monomeric C-reactive protein in patients with myocardial infarction. Cardiovasc Res 96(1):64–72. https://doi.org/10.1093/cvr/cvs237
Cordazzo C, Petrini S, Neri T, Lombardi S, Carmazzi Y, Pedrinelli R, Paggiaro P, Celi A (2014) Rapid shedding of proinflammatory microparticles by human mononuclear cells exposed to cigarette smoke is dependent on Ca2+ mobilization. Inflamm Res 63(7):539–547. https://doi.org/10.1007/s00011-014-0723-7
Terrisse AD, Puech N, Allart S, Gourdy P, Xuereb JM, Payrastre B, Sie P (2010) Internalization of microparticles by endothelial cells promotes platelet/endothelial cell interaction under flow. J Thromb Haemost 8(12):2810–2819. https://doi.org/10.1111/j.1538-7836.2010.04088.x
Christopher AF, Kaur RP, Kaur G, Kaur A, Gupta V, Bansal P (2016) MicroRNA therapeutics: discovering novel targets and developing specific therapy. Perspect Clin Res 7(2):68–74. https://doi.org/10.4103/2229-3485.179431
Cheng L, Sharples RA, Scicluna BJ, Hill AF (2014) Exosomes provide a protective and enriched source of miRNA for biomarker profiling compared to intracellular and cell-free blood. J Extracell Ves 3(1):23743. https://doi.org/10.3402/jev.v3.23743
Kaur A, Mackin ST, Schlosser K, Wong FL, Elharram M, Delles C, Stewart DJ, Dayan N, Landry T, Pilote L (2019) Systematic review of microRNA biomarkers in acute coronary syndrome and stable coronary artery disease. Cardiovasc Res. https://doi.org/10.1093/cvr/cvz302
This work was supported by the German Research Foundation (DFG) and by the National Health and Medical Research Council (NHMRC) of Australia.
Conflict of interest
The authors declare no conflicts of interest.
All animal studies were approved by the ethics committee of the University of Freiburg Medical Center (Ethics No. X17/02R).
Consent for publication
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Siegel, P.M., Schmich, J., Barinov, G. et al. Cardiomyocyte microvesicles: proinflammatory mediators after myocardial ischemia?. J Thromb Thrombolysis (2020). https://doi.org/10.1007/s11239-020-02156-x
- C-reactive protein
- Myocardial infarction