Biochemical changes in endothelial cell monolayers induced by fibrin deposition in vitro.

Schleef, Raymond R.; Birdwell, Charles R.

doi:10.1161/01.atv.4.1.14

Cited by 28 publications

(21 citation statements)

References 37 publications

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“…They found a major component of fibrin II (lacking both FPA and FPB) in all plaques and mural thrombi, and this was also demonstrated immunohistochemically. 38 Their findings also agreed with ours in showing a consistent presence of fragment X. Fragment X may be derived from flbrinogen or from fibrin I monomer 36 ; all our fragment X bands (band d) contained FPA, indicating a fibrinogen precursor, but this does not preclude the possibility of comigration with X derived from fibrin, and the difference in molecular weight of 1800 is not measurable in our system.…”

Section: Discussionsupporting

confidence: 89%

Fate of fibrinogen in human arterial intima.

et al. 1990

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Fibrinogen and flbrinogen/flbrin-related antigen (total FRA) was measured In human normal Intima and different types of atherosclerotic lesions and mural thrombi. The amount showed marked variation between groups of tissue samples, but within each group there was a significant correlation between levels of total FRA and low density llpoproteln (LDL), suggesting that some common factor must Influence their influx or retention. The total FRA were analyzed by gradient sodium dodecyl sutfate polyacrylamlde gel electrophoresis and ImmunoblottJng with antisera to whole fibrinogen and fragments D and E, and fibrinopeptlde A (FPA). All intimal samples (but not thrombi) contained fragment X, the first product of plasmln digestion of fibrinogen, but fragment Y was present In only half the samples, and no core-fragment E containing FPA was detected in any sample, suggesting that flbr/nogenolysis Is limited. By contrast, all samples contained fragment E, which was negative for FPA, so presumably derived from fibrin; they also contained fragments D-dlmer and DY, which are characteristic degradation products of cross-linked fibrin. There were no differences between samples obtained during reconstructive 1 The early proliferative lesions appear first as small translucent elevations but may progress to massive translucent proliferate mounds, up to 2500 tun In thickness, which are particularly characterized by their high water content; this gives them a soft and wobbly texture, and they are described as gelatinous lesions. 23 In a high proportion of these lesions, neither extracellular lipid nor lipid within fat-filled SMC or macrophages can be demonstrated in frozen microscopical sections, suggesting that lipid deposition cannot be the factor initiating SMC proliferation. These lesions contain large amounts of plasma macromolecules, including low density lipoprotein (LDL), fibrinogen and fibrin-related antigens (FRA), other components of the hemostatic system, and insoluble fibrins** In a preliminary study of the FRA fraction, Smith and Walker 6 found that about naff was comprised of intact fibrinogen, but the remainder consisted of fragments, mainly of high molecular mass, which showed characteristic patterns in different types of lesions. Different fibrin or fibrinogen degradation products have a wide range of pharmacobiological activities, including stimulation of growth of cultured cells; modification of clotting, vascular all of which might influence atherogenesis. In the present study we have attempted a detailed identification of the FRA present in lesions to relate them to particular biological activities, and, by determining if they are derived from fibrinogen or fibrin, to gain a better insight Into the handling of hemostatic factors in the arterial wall. Previously, we isolated the FRA from intimal extracts by affinity chromatography on Sepharose-antjfibrinogen columns, followed by elution with 8 M urea and separation by sodium dodecyl sutfate polyacrylamide gel electrophoresis (SDS-PAGE) and staining with Coomassie ...

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Section: Discussionsupporting

confidence: 89%

Fate of fibrinogen in human arterial intima.

et al. 1990

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“…Furthermore, direct contact of endothelial cells with fibrin has been shown to modulate a number of phenotypic properties, including loss of organization, with severing of cell-cell contacts and retraction of individual endothelial cells. [33][34][35] In addition to enhancing the production of PAI-1 in the single layer of endothelial cells lining the vessel wall, the deposition of a thromboembolus within a pulmonary vessel results in the generation of a new interface, which is subsequently penetrated by cells that are involved in the organization process. Our observation of not only elevated PAI-1 but also enhanced u-PA expression in areas of initial thrombus organization and increased cellular proliferation extends the data of Pepper and coworkers, 36,37 in which the expression of these two proteins was observed to be upregulated in proliferating and migrating cells.…”

Section: Discussionmentioning

confidence: 99%

“…Parallel sections were analyzed by using a sense probe as the control for nonspecific hybridization. For quantitative analysis of PAI-1 mRNA at the single cell level, the 1085-bp fragment of PAI-1 in pGEM-3Z was labeled with 35 S-UTP and used for in situ hybridization experiments as described previously. 8 Specimens were analyzed at 10 weeks' exposure using combined light/epiluminescence microscopy to permit simultaneous visualization of the sample and exposed silver grains.…”

Section: Table 1 Application Of a Panel Of Antibodies For The Analysmentioning

confidence: 99%

Elevated Expression of Urokinase-like Plasminogen Activator and Plasminogen Activator Inhibitor Type 1 During the Vascular Remodeling Associated With Pulmonary Thromboembolism

Lang

Moser

Schleef

1998

ATVB

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Abstract-Information is lacking on the mechanisms involved in the organization, resolution, and repair of the vascular lumen after acute pulmonary thromboembolism. Because recent data suggest that the balance between plasminogen activators (PAs) and type 1 plasminogen activator inhibitor (PAI-1) plays a role in regulating cell migration within the extracellular matrix, we investigated the expression of these molecules by immunohistochemical and in situ hybridization analysis of pulmonary artery specimens from patients suffering fatal pulmonary embolism. The data were compared with the expression of these molecules in both patients' noninvolved pulmonary arteries and organ donor pulmonary arteries. Regions of initial organization and vascular remodeling were identified by a modified trichrome stain and by the presence of proliferating cell nuclear antigen (PCNA), a cell marker of proliferation. Staining for tissue-type PA antigen was low to undetectable in endothelial cells directly in contact with the fibrin-platelet thromboembolus and in areas in which the endothelial cell lining was replaced by cell growth into the thrombus. Urokinase-like PA (u-PA) expression was detected in mononuclear cells within the thrombus in the initial phase of thromboembolism and within cells migrating into the thrombus during the later stages of organization. PAI-1 expression was elevated in the monolayer of endothelial cells underlying the fresh platelet-fibrin thromboembolus and in a PCNA-positive cell population present between the pulmonary arterial intima and the thromboembolus that represents early organization. Increased expression of PAI-1 may play a role in inhibiting proteolysis and fostering the localization of the acute fibrin-platelet thrombus to the vascular wall, which is followed by the upregulation of u-PA in migrating cells during the reorganization process. (Arterioscler Thromb Vasc Biol. 1998;18:808-815.)Key Words: thrombus organization Ⅲ plasminogen activators Ⅲ plasminogen activator inhibitor type 1 Ⅲ pulmonary thromboembolism R ecognized venous thromboembolism, comprising both pulmonary embolism and deep venous thrombosis, leads to sudden death in about 10% of patients, accounting for about 300 000 clinical episodes and 50 000 deaths per year in the United States.1,2 The origin of thrombi embolizing to the lungs is in the great majority of cases the deep veins of the legs, particularly iliac, femoral, and popliteal veins.3 The morphological features of a fresh thromboembolus are similar to those of a nondetached thrombus, with alternating layers of platelets, fibrin, and erythrocytes.3 In addition to obstruction of the pulmonary trunk and/or both main pulmonary arteries, pulmonary embolism may progress slowly until the vascular bed has been reduced by multiple small pulmonary thromboemboli. In the latter situation, a single fresh thromboembolus suffices to cause acute right ventricular failure.Resolution of pulmonary thromboemboli is usually derived by mechanical fragmentation, endogenous thrombolysis, or repa...

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“…Initiation of endothelial cell growth was observed when collagen gels 17 induced disorganization of the monolayers, but no growth-promoting effect for fibrin was detected. 37 Additional support for the concept that disorganization of endothelial cells is the initial step required for the recanalization of an occluding thrombus was provided by the recent finding that lumina form and new vessels develop when capillary endothelial cells were covered with collagen gels. 38 …”

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confidence: 99%

The mechanism of fibrin-induced disorganization of cultured human endothelial cell monolayers.

Weimar¹,

Delvos²

1986

Arteriosclerosis

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Deposition of polymerizing fibrin on the vascular endothelium is the final event in intravascular coagulation. Exposure of fibrin clots to confluent monolayers of cultured human endothelial cells for 4 to 24 hours resulted in the disappearance of their normal cobblestone morphology and in the formation of endothelial cell aggregates. The present study was designed to evaluate the conditions and structural requirements of the fibrin clot for the induction of disorganization. Even after harsh treatment with denaturing agents or loading with large amounts of fibrinogen antibodies, polymerized fibrin always induced disorganization of the monolayers. In contrast, soluble fibrin that was kept in solution by either fibrinogen, fragment D-cate, or the tetrapeptide Gly-Pro-Arg-Pro did not cause any alteration of the monolayers. The fibrinogen degradation product D-cate (M r 94,000) itself had no microscopically detectable influence on the monolayer structure. In the absence of fibrin, the effect of thrombin on endothelial cells was found to be distinct from that induced by fibrin; however, the exposure of pieces of glass coverslips caused alterations in morphology indistinguishable from the fibrin-induced disorganization of the monolayer. Experiments using protein-coated polyester films indicated that the ability of the endothelial cells to attach to the overlying material, independent of its chemical structure, is the prerequisite for the induction of disorganization, but not a defined component of the fibrin molecule. Disorganization of vascular endothelium in vivo might be important for the organization and revascularization of an occluding thrombus. (Arteriosclerosis 6:139-145, March/April 1986) T he vascular endothelium contributes anticoagulant as well as procoagulant activities to the regulation of the coagulation system. A nonthrombogenic surface and the expression of fibrinolytic activities by the endothelial cells guarantee the patency of the vascular tree. Surface-bound heparan sulphate, which accelerates the inactivation of thrombin by enhancing the formation of the antithrombin Ill/thrombin complex, 1 and the membrane cofactor thrombomodulin, which facilitates the activation of protein C in conjunction with thrombin, 2 are important surface-mediated mechanisms of the vessel wall that are anticoagulant in nature. In addition, the synthesis and release of prostacyclin 3 and of tissue plasminogen activators 4 account for the inhibition of platelet activation and the continuous lysis of small amounts of fibrin. On the other hand, the synthesis of facator VIIIR:Ag, 56 specific binding of factors IX/IXa and X/Xa, 7 ' 8 and the induction of tissue factor production in the This study was supported by the Stiftung Volkswagenwerk, Hannover, West Germany.Part of this work was presented at the 10th International Congress on Thrombosis and Haemostasis, San Diego, California, in 1985 and is published in abstract form (Thromb Haemostas 1985;54:194).Address for reprints: Dr. Ulrich Delvos, Clinical Research Unit for Blood ...

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Biochemical changes in endothelial cell monolayers induced by fibrin deposition in vitro.

Abstract: Endothelial cells In vivo come

Cited by 28 publications

References 37 publications

Fate of fibrinogen in human arterial intima.

Fate of fibrinogen in human arterial intima.

Elevated Expression of Urokinase-like Plasminogen Activator and Plasminogen Activator Inhibitor Type 1 During the Vascular Remodeling Associated With Pulmonary Thromboembolism

The mechanism of fibrin-induced disorganization of cultured human endothelial cell monolayers.

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