title: Oxidized hemoglobin is an endogenous proinflammatory agonist that targets vascular endothelial cells.
abstract: Several pathologic conditions are associated with hemolysis, i.e. release of ferrous (Fe(II)) hemoglobin from red blood cells. Oxidation of cell-free hemoglobin produces (Fe(III)) methemoglobin. More extensive oxidation produces (Fe(III)/Fe(IV) O) ferryl hemoglobin. Both cell-free methemoglobin and ferryl hemoglobin are thought to contribute to the pathogenesis of hemolytic disorders. We show hereby that ferryl hemoglobin, but not hemoglobin or methemoglobin, acts as a potent proinflammatory agonist that induces vascular endothelial cells in vitro to rearrange the actin cytoskeleton, forming intercellular gaps and disrupting the integrity of the endothelial cell monolayer. Furthermore, ferryl hemoglobin induces the expression of proinflammatory genes in endothelial cells in vitro, e.g. E-selectin, Icam-1, and Vcam-1, through the activation of the nuclear factor kappaB family of transcription factors. This proinflammatory effect, which requires actin polymerization, involves the activation of the c-Jun N-terminal kinase and the p38 mitogen-activated protein kinase signal transduction pathways. When administered to naïve mice, ferryl hemoglobin induces the recruitment of polymorphonuclear cells, demonstrating that it acts as a proinflammatory agonist in vivo. In conclusion, oxidized hemoglobin, i.e. ferryl hemoglobin, acts as a proinflammatory agonist that targets vascular endothelial cells.
authors.affiliation: Instituto Gulbenkian de Ciência, Oeiras 2780-156, Portugal.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Silva
authors.forename: Viktoria
authors.initials: V
authors.lastname: Jeney
authors.forename: Angelo
authors.initials: A
authors.lastname: Chora
authors.forename: Rasmus
authors.initials: R
authors.lastname: Larsen
authors.forename: Jozsef
authors.initials: J
authors.lastname: Balla
authors.forename: Miguel P
authors.initials: MP
authors.lastname: Soares
mesh_terms: D000818:Animals; D015815:Cell Adhesion Molecules / Q000096:biosynthesis; D002460:Cell Line; D042783:Endothelial Cells / Q000378:metabolism*; D005786:Gene Expression Regulation; D006454:Hemoglobins / Q000378:metabolism* / Q000494:pharmacology; D006461:Hemolysis; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism* / Q000494:pharmacology; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D020935:MAP Kinase Signaling System; D008706:Methemoglobin / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D016328:NF-kappa B / Q000378:metabolism; D009504:Neutrophils / Q000378:metabolism; D010108:Oxyhemoglobins / Q000378:metabolism* / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D015815:Cell Adhesion Molecules; D006454:Hemoglobins; D018836:Inflammation Mediators; D016328:NF-kappa B; D010108:Oxyhemoglobins; C517773:ferrylhemoglobin; D008706:Methemoglobin; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases
references.citation: J Clin Invest. 1983 Jun;71(6):1893-6
references.pmid: 6345590
references.citation: Methods Enzymol. 1994;231:490-6
references.pmid: 8041270
references.citation: Biochem Soc Trans. 2002 Aug;30(4):745-8
references.pmid: 12196184
references.citation: J Exp Med. 2000 Oct 2;192(7):1015-26
references.pmid: 11015442
references.citation: Proc Natl Acad Sci U S A. 1968 Oct;61(2):748-55
references.pmid: 4386763
references.citation: J Immunol. 2006 Aug 1;177(3):1894-903
references.pmid: 16849502
references.citation: Annu Rev Med. 1996;47:315-31
references.pmid: 8712785
references.citation: Blood. 2008 Nov 15;112(10):3927-38
references.pmid: 18988877
references.citation: J Biol Chem. 2007 Feb 16;282(7):4894-4907
references.pmid: 17178725
references.citation: J Biol Chem. 2001 Jun 29;276(26):24129-36
references.pmid: 11294851
references.citation: Nature. 1997 Jul 24;388(6640):394-7
references.pmid: 9237759
references.citation: Nat Rev Mol Cell Biol. 2004 May;5(5):392-401
references.pmid: 15122352
references.citation: Arterioscler Thromb Vasc Biol. 1996 Aug;16(8):1047-51
references.pmid: 8696945
references.citation: Am J Physiol Cell Physiol. 2003 Nov;285(5):C1036-46
references.pmid: 12839837
references.citation: J Biol Chem. 1999 Jul 2;274(27):18916-24
references.pmid: 10383389
references.citation: Science. 1988 Oct 28;242(4878):540-6
references.pmid: 3140380
references.citation: Blood. 1994 May 15;83(10):2860-5
references.pmid: 8180381
references.citation: Hum Pathol. 2002 Jan;33(1):125-30
references.pmid: 11823983
references.citation: Nature. 2006 Mar 9;440(7081):237-41
references.pmid: 16407889
references.citation: Science. 1995 Mar 10;267(5203):1485-8
references.pmid: 7878466
references.citation: J Biol Chem. 1997 Jan 10;272(2):843-51
references.pmid: 8995372
references.citation: Free Radic Biol Med. 2008 Oct 15;45(8):1150-8
references.pmid: 18708138
references.citation: Cell. 1996 Sep 6;86(5):787-98
references.pmid: 8797825
references.citation: J Clin Invest. 1999 Jan;103(1):129-35
references.pmid: 9884342
references.citation: Annu Rev Immunol. 2009;27:229-65
references.pmid: 19302040
references.citation: J Exp Med. 1996 Mar 1;183(3):1013-22
references.pmid: 8642242
references.citation: Nature. 2003 Oct 2;425(6957):516-21
references.pmid: 14520412
references.citation: Blood. 2002 Aug 1;100(3):879-87
references.pmid: 12130498
references.citation: Nature. 1958 Nov 15;182(4646):1373
references.pmid: 13600326
references.citation: Nat Immunol. 2008 Aug;9(8):857-65
references.pmid: 18604209
references.citation: Anal Biochem. 1992 Dec;207(2):311-6
references.pmid: 1481986
references.citation: Blood. 2001 Sep 15;98(6):1802-11
references.pmid: 11535514
references.citation: JAMA. 2005 Apr 6;293(13):1653-62
references.pmid: 15811985
references.citation: J Immunol. 1998 Nov 1;161(9):4572-82
references.pmid: 9794384
references.citation: J Biol Chem. 2008 Nov 7;283(45):30780-7
references.pmid: 18728007
references.citation: J Biol Chem. 2005 Jan 7;280(1):244-52
references.pmid: 15516339
references.citation: Free Radic Biol Med. 1998 Jan 15;24(2):269-79
references.pmid: 9433902
references.citation: Nature. 1993 Sep 9;365(6442):182-5
references.pmid: 8371761
references.citation: Nat Rev Immunol. 2007 Oct;7(10):803-15
references.pmid: 17893694
references.citation: Arch Biochem Biophys. 1990 Mar;277(2):402-9
references.pmid: 2155581
references.citation: Proc Natl Acad Sci U S A. 1997 Sep 30;94(20):10919-24
references.pmid: 9380735
references.citation: Trends Mol Med. 2009 Feb;15(2):50-8
references.pmid: 19162549
references.citation: J Immunol. 2007 Dec 1;179(11):7840-51
references.pmid: 18025230
references.citation: J Biol Chem. 1982 Mar 25;257(6):2947-53
references.pmid: 6277918
references.citation: J Immunol. 2004 Mar 15;172(6):3553-63
references.pmid: 15004156
references.citation: J Biol Chem. 1994 May 27;269(21):14869-71
references.pmid: 8195116
references.citation: Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9285-9
references.pmid: 8415693
references.citation: J Cell Biol. 1987 Oct;105(4):1473-8
references.pmid: 3312229
references.citation: EMBO J. 1994 Nov 15;13(22):5433-41
references.pmid: 7957109
references.citation: Infect Immun. 1997 Apr;65(4):1258-66
references.pmid: 9119460
references.citation: Mol Cell. 2003 Oct;12(4):829-39
references.pmid: 14580335
references.citation: Free Radic Biol Med. 2005 Nov 1;39(9):1216-28
references.pmid: 16214037
references.citation: Genes Dev. 1995 Jul 1;9(13):1586-97
references.pmid: 7628694
references.citation: Am J Physiol. 1998 Sep;275(3):H1046-53
references.pmid: 9724312
references.citation: Nat Cell Biol. 2000 Jun;2(6):376-8
references.pmid: 10854330
references.citation: J Mol Med (Berl). 2008 Oct;86(10):1097-111
references.pmid: 18641963
references.citation: Science. 1987 Feb 27;235(4792):1043-6
references.pmid: 3029864
references.citation: Nat Rev Immunol. 2004 Jun;4(6):469-78
references.pmid: 15173835
references.citation: J Clin Invest. 1995 Apr;95(4):1782-8
references.pmid: 7535798
references.citation: J Immunol. 1999 Apr 1;162(7):3749-52
references.pmid: 10201887
references.citation: J Biol Chem. 2006 Aug 4;281(31):21728-21734
references.pmid: 16751193
references.citation: J Biol Chem. 1997 Mar 14;272(11):7114-21
references.pmid: 9054405
references.citation: Nat Rev Mol Cell Biol. 2007 Jan;8(1):49-62
references.pmid: 17183360
references.citation: Nature. 2001 Jan 11;409(6817):198-201
references.pmid: 11196644
references.citation: Trends Immunol. 2003 Aug;24(8):449-55
references.pmid: 12909459
references.citation: Blood. 2001 Dec 15;98(13):3693-8
references.pmid: 11739174