Background Hepatic ischemia-reperfusion (I/R) is a well-studied model of liver injury and has demonstrated a biphasic injury followed by recovery and regeneration. Microparticles (MPs) are a developing field of study and these small membrane bound vesicles have been shown to have effector function in other physiologic and pathologic states. This study was designed to quantify the levels of MPs from various cell origins–platelets, neutrophils, and endolethial cells–following hepatic ischemia-reperfusion injury. Methods A murine model was used with mice undergoing 90 minutes of partial hepatic ischemia followed by various times of reperfusion. Following reperfusion, plasma samples were taken and MPs of various cell origins were labeled and levels were measured using flow cytometry. Additionally, cell specific MPs were further assessed by Annexin V, which stains for the presence of phosphatidylserine, a cell surface marker linked to apoptosis. Statistical analysis was performed using one-way analysis of variance with subsequent Student-Newman-Keuls test with data presented as the mean and standard error of the mean. Results MPs from varying sources show an increase in circulating levels following hepatic I/R injury. However, the timing of the appearance of different MP subtypes differs for each cell type. Platelet and neutrophil-derived MP levels demonstrated an acute elevation following injury whereas endothelial-derived MP levels demonstrated a delayed elevation. Conclusion This is the first study to characterize circulating levels of cell-specific MPs after hepatic I/R injury and suggests that MPs derived from platelets and neutrophils serve as markers of inflammatory injury and may be active participants in this process. In contrast, MPs derived from endothelial cells increase after the injury response during the reparative phase and may be important in angiogenesis that occurs in the regenerating liver.
Liu DL, Jeppsson B, Hakansson CH, Odselius R (1996) Multiple-system organ damage resulting from prolonged hepatic inflow interruption. Arch Surg 131: 442–447.
[3]
Huguet C, Gavelli A, Bona S (1994) Hepatic resection with ischemia of the liver exceeding one hour. J Am Coll Surg 178: 454–458.
[4]
Lemasters JJ, Thurman RG (1997) Reperfusion injury after liver preservation for transplantation. Annu Rev Pharmacol Toxicol 37: 327–338.
[5]
Jaeschke H, Farhood A, Smith CW (1990) Neutrophils contribute to ischemia/reperfusion injury in rat liver in vivo. FASEB J 4: 3355–3359.
[6]
Jaeschke H (2006) Mechanisms of Liver Injury. II. Mechanisms of neutrophil-induced liver cell injury during hepatic ischemia-reperfusion and other acute inflammatory conditions. Am J Physiol Gastrointest Liver Physiol 290: G1083–1088.
[7]
Lentsch AB, Kato A, Yoshidome H, McMasters KM, Edwards MJ (2000) Inflammatory mechanisms and therapeutic strategies for warm hepatic ischemia/reperfusion injury. Hepatology 32: 169–173.
[8]
Husted TL, Lentsch AB (2006) The role of cytokines in pharmacological modulation of hepatic ischemia/reperfusion injury. Curr Pharm Des 12: 2867–2873.
[9]
Jaeschke H, Bautista AP, Spolarics Z, Spitzer JJ (1992) Superoxide generation by neutrophils and Kupffer cells during in vivo reperfusion after hepatic ischemia in rats. J Leukoc Biol 52: 377–382.
[10]
Jaeschke H, Farhood A (1991) Neutrophil and Kupffer cell-induced oxidant stress and ischemia-reperfusion injury in rat liver. Am J Physiol 260: G355–362.
[11]
Morel O, Jesel L, Freyssinet JM, Toti F (2011) Cellular mechanisms underlying the formation of circulating microparticles. Arterioscler Thromb Vasc Biol 31: 15–26.
[12]
Azevedo LC, Pedro MA, Laurindo FR (2007) Circulating microparticles as therapeutic targets in cardiovascular diseases. Recent Pat Cardiovasc Drug Discov 2: 41–51.
[13]
Morel O, Morel N, Jesel L, Freyssinet JM, Toti F (2011) Microparticles: a critical component in the nexus between inflammation, immunity, and thrombosis. Semin Immunopathol 33: 469–486.
[14]
Lee TH, D’Asti E, Magnus N, Al-Nedawi K, Meehan B, et al. (2011) Microvesicles as mediators of intercellular communication in cancer–the emerging science of cellular ‘debris’. Semin Immunopathol 33: 455–467.
[15]
Hunter MP, Ismail N, Zhang X, Aguda BD, Lee EJ, et al. (2008) Detection of microRNA expression in human peripheral blood microvesicles. PLoS One 3: e3694.
[16]
George JN, Thoi LL, McManus LM, Reimann TA (1982) Isolation of human platelet membrane microparticles from plasma and serum. Blood 60: 834–840.
[17]
Meziani F, Delabranche X, Asfar P, Toti F (2010) Bench-to-bedside review: circulating microparticles–a new player in sepsis? Crit Care 14: 236.
[18]
Ogura H, Kawasaki T, Tanaka H, Koh T, Tanaka R, et al. (2001) Activated platelets enhance microparticle formation and platelet-leukocyte interaction in severe trauma and sepsis. J Trauma 50: 801–809.
[19]
Densmore JC, Signorino PR, Ou J, Hatoum OA, Rowe JJ, et al. (2006) Endothelium-derived microparticles induce endothelial dysfunction and acute lung injury. Shock 26: 464–471.
[20]
Lentsch AB, Yoshidome H, Cheadle WG, Miller FN, Edwards MJ (1998) Chemokine involvement in hepatic ischemia/reperfusion injury in mice: roles for macrophage inflammatory protein-2 and KC. Hepatology 27: 1172–1177.
[21]
Prakash PS, Caldwell CC, Lentsch AB, Pritts TA, Robinson BR (2012) Human microparticles generated during sepsis in patients with critical illness are neutrophil-derived and modulate the immune response. J Trauma Acute Care Surg 73: 401–406 discussion 406–407.
[22]
Dalli J, Jones CP, Cavalcanti DM, Farsky SH, Perretti M, et al. (2012) Annexin A1 regulates neutrophil clearance by macrophages in the mouse bone marrow. FASEB J 26: 387–396.
[23]
Nagendra S, Schlueter AJ (2004) Absence of cross-reactivity between murine Ly-6C and Ly-6G. Cytometry A 58: 195–200.
[24]
Mostefai HA, Meziani F, Mastronardi ML, Agouni A, Heymes C, et al. (2008) Circulating microparticles from patients with septic shock exert protective role in vascular function. Am J Respir Crit Care Med 178: 1148–1155.
[25]
Ogura H, Tanaka H, Koh T, Fujita K, Fujimi S, et al. (2004) Enhanced production of endothelial microparticles with increased binding to leukocytes in patients with severe systemic inflammatory response syndrome. J Trauma 56: 823–830 discussion 830–821.
[26]
Janiszewski M, Do Carmo AO, Pedro MA, Silva E, Knobel E, et al. (2004) Platelet-derived exosomes of septic individuals possess proapoptotic NAD(P)H oxidase activity: A novel vascular redox pathway. Crit Care Med 32: 818–825.
[27]
Forlow SB, McEver RP, Nollert MU (2000) Leukocyte-leukocyte interactions mediated by platelet microparticles under flow. Blood 95: 1317–1323.
[28]
Barry OP, Pratico D, Savani RC, FitzGerald GA (1998) Modulation of monocyte-endothelial cell interactions by platelet microparticles. J Clin Invest 102: 136–144.
[29]
Brown GT, McIntyre TM () Lipopolysaccharide signaling without a nucleus: kinase cascades stimulate platelet shedding of proinflammatory IL-1beta-rich microparticles. J Immunol 186: 5489–5496.
[30]
Jy W, Mao WW, Horstman L, Tao J, Ahn YS (1995) Platelet microparticles bind, activate and aggregate neutrophils in vitro. Blood Cells Mol Dis 21: 217–231 discussion 231a.
[31]
Yadav SS, Howell DN, Steeber DA, Harland RC, Tedder TF, et al. (1999) P-Selectin mediates reperfusion injury through neutrophil and platelet sequestration in the warm ischemic mouse liver. Hepatology. 29: 1494–1502.
[32]
Sindram D, Porte RJ, Hoffman MR, Bentley RC, Clavien PA (2000) Platelets induce sinusoidal endothelial cell apoptosis upon reperfusion of the cold ischemic rat liver. Gastroenterology 118: 183–191.
[33]
Lindemann S, Tolley ND, Dixon DA, McIntyre TM, Prescott SM, et al. (2001) Activated platelets mediate inflammatory signaling by regulated interleukin 1beta synthesis. J Cell Biol 154: 485–490.
[34]
Gasser O, Schifferli JA (2005) Microparticles released by human neutrophils adhere to erythrocytes in the presence of complement. Exp Cell Res 307: 381–387.
[35]
Mesri M, Altieri DC (1998) Endothelial cell activation by leukocyte microparticles. J Immunol 161: 4382–4387.
[36]
Mesri M, Altieri DC (1999) Leukocyte microparticles stimulate endothelial cell cytokine release and tissue factor induction in a JNK1 signaling pathway. J Biol Chem 274: 23111–23118.
[37]
Faure V, Dou L, Sabatier F, Cerini C, Sampol J, et al. (2006) Elevation of circulating endothelial microparticles in patients with chronic renal failure. J Thromb Haemost 4: 566–573.
[38]
Werner N, Wassmann S, Ahlers P, Kosiol S, Nickenig G (2006) Circulating CD31+/annexin V+ apoptotic microparticles correlate with coronary endothelial function in patients with coronary artery disease. Arterioscler Thromb Vasc Biol 26: 112–116.
[39]
Dignat-George F, Boulanger CM (2011) The many faces of endothelial microparticles. Arterioscler Thromb Vasc Biol 31: 27–33.
[40]
Leroyer AS, Rautou PE, Silvestre JS, Castier Y, Leseche G, et al. (2008) CD40 ligand+ microparticles from human atherosclerotic plaques stimulate endothelial proliferation and angiogenesis a potential mechanism for intraplaque neovascularization. J Am Coll Cardiol 52: 1302–1311.
