S-Nitrosoglutathione Acts as a Small Molecule Modulator of Human Fibrin Clot Architecture 英文参考文献.docVIP

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S-Nitrosoglutathione Acts as a Small Molecule Modulator of Human Fibrin Clot Architecture 英文参考文献.doc

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S-Nitrosoglutathione Acts as a Small Molecule Modulator of Human Fibrin Clot Architecture 英文参考文献

S-NitrosoglutathioneActsasaSmallMoleculeModulator ofHumanFibrinClotArchitecture RyonM.Bateman1,2*,ChristopherG.Ellis3,MakotoSuematsu1,KeithR.Walley2 1DepartmentofBiochemistry,SchoolofMedicine,KeioUniversity,Tokyo,Japan,2HeartLungInstitute,UniversityofBritishColumbia,Vancouver,Canada,3Department ofMedicalBiophysics,UniversityofWesternOntario,London,Canada Abstract Background:Alteredfibrinclotarchitectureisincreasinglyassociatedwithcardiovasculardiseases;yet,littleisknownabout how fibrin networks are affected by small molecules that alter fibrinogen structure. Based on previous evidence that S- nitrosoglutathione(GSNO)altersfibrinogen secondary structureandfibrinpolymerizationkinetics, wehypothesized that GSNOwouldalterfibrinmicrostructure. Methodology/Principal Findings: Accordingly, we treated human platelet-poor plasma with GSNO (0.01–3.75mM) and imagedthrombininducedfibrinnetworksusingmultiphotonmicroscopy.Usingcustomdesignedcomputersoftware,we analyzed fibrin microstructure for changes in structural features including fiber density, diameter, branch point density, crossing fibers and void area. We report for the first time that GSNO dose-dependently decreased fibrin density until completenetworkinhibitionwasachieved.AtlowdoseGSNO,fiberdiameterincreased25%,maintainingclotvoidvolume at approximately 70%. However, at high dose GSNO, abnormal irregularly shaped fibrin clusters with high fluorescence intensity cores were detected and clot void volume increased dramatically. Notwithstanding fibrin clusters, the clot remainedstable,asfiberbranchingwasinsensitivetoGSNOandtherewasnoevidenceoffibermotionwithinthenetwork. Moreover, at the highest GSNO dose tested, we observed for the first time, that GSNO induced formation of fibrin agglomerates. Conclusions/Significance: Taken together, low dose GSNO modulated fibrin microstructure generating coarse fibrin networkswiththickerfibers;however,higherdosesofGSNOinducedabnormalfibrinstructuresandfibrinagglomerates. SinceGSN