Fibrin γ/γ' influences the secretion of fibrinolytic components and clot structure
Abstract Background In healthy subjects fibrinogen γ/γ‘ circulates at 8–15% of the total plasma fibrinogen concentration. Elevated levels of this variant have been associated with arterial thrombosis, and its diminution with venous thrombosis. The aims of the present work were to analyze the structu...
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| Format: | Article |
| Language: | English |
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BMC
2019-11-01
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| Series: | BMC Molecular and Cell Biology |
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| Online Access: | http://link.springer.com/article/10.1186/s12860-019-0233-0 |
| _version_ | 1818211941464670208 |
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| author | Miriam Cantero Héctor Rojas Eduardo Anglés-Cano Rita Marchi |
| author_facet | Miriam Cantero Héctor Rojas Eduardo Anglés-Cano Rita Marchi |
| author_sort | Miriam Cantero |
| collection | DOAJ |
| description | Abstract Background In healthy subjects fibrinogen γ/γ‘ circulates at 8–15% of the total plasma fibrinogen concentration. Elevated levels of this variant have been associated with arterial thrombosis, and its diminution with venous thrombosis. The aims of the present work were to analyze the structure of the fibrin network formed on the top of human dermal microvascular endothelial cells (HMEC-1) at different fibrinogen γ/γ‘ concentrations, as well as its influence on the secretion of fibrinolytic components. The kinetics of fibrin polymerization on top of HMEC-1 cells with 3, 10, and 30% fibrinogen γ/γ‘ was followed at 350 nm. The secretion of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor type 1 (PAI 1) by HMEC-1 were measured in the supernatant and cell lysates, after incubation with 1 nM thrombin, fibrin with 3, and 30% fibrinogen γ/γ‘, using commercial kits. The influence of fibrinogen γ/γ‘ on fibrin structure on the surface of the HMEC-1 was followed with laser scanning confocal microscopy (LSCM). Results The kinetics of fibrin formation on HMEC-1 with 3 and 10% fibrinogen γ/γ‘ were similar. However, with 30% fibrinogen γ/γ‘ both the slope and final turbity were approximately 50% less. The LSCM images showed the dramatic effects of increasing fibrinogen γ/γ‘ from 3 to 30%. The uPA and PAI 1 concentrations in culture supernatants HMEC-1 cells treated with thrombin or 30% γ/γ‘ fibrin were two-fold increased as compared to basal culture supernatants and 3% γ/γ‘ fibrin-treated HMEC-1. In all stimulatory conditions the intracellular concentration of uPA was higher than in supernatants. In contrast, the intracellular PAI 1 concentration was decreased as compared to that measured in the supernatant, including the basal condition. Conclusion A concentration of 30% fibrin γ/γ‘ alter drastically fibrin structure on the cell surface and affects the secretion of uPA and PAI 1 through its capacity to bind thrombin. |
| first_indexed | 2024-12-12T05:40:30Z |
| format | Article |
| id | doaj.art-f41c188713ae4a9e9c2a0f0872ebb9d1 |
| institution | Directory Open Access Journal |
| issn | 2661-8850 |
| language | English |
| last_indexed | 2024-12-12T05:40:30Z |
| publishDate | 2019-11-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Molecular and Cell Biology |
| spelling | doaj.art-f41c188713ae4a9e9c2a0f0872ebb9d12022-12-22T00:35:57ZengBMCBMC Molecular and Cell Biology2661-88502019-11-012011910.1186/s12860-019-0233-0Fibrin γ/γ' influences the secretion of fibrinolytic components and clot structureMiriam Cantero0Héctor Rojas1Eduardo Anglés-Cano2Rita Marchi3Departamento de Bioquímica, Universidad de CórdobaInstituto de Inmunología, Universidad Central de VenezuelaUniversité de Paris, Innovative Therapies in Haemostasis, INSERMCentro de Medicina Experimental, Laboratorio Biología del Desarrollo de la Hemostasia, Instituto Venezolano de Investigaciones CientíficasAbstract Background In healthy subjects fibrinogen γ/γ‘ circulates at 8–15% of the total plasma fibrinogen concentration. Elevated levels of this variant have been associated with arterial thrombosis, and its diminution with venous thrombosis. The aims of the present work were to analyze the structure of the fibrin network formed on the top of human dermal microvascular endothelial cells (HMEC-1) at different fibrinogen γ/γ‘ concentrations, as well as its influence on the secretion of fibrinolytic components. The kinetics of fibrin polymerization on top of HMEC-1 cells with 3, 10, and 30% fibrinogen γ/γ‘ was followed at 350 nm. The secretion of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor type 1 (PAI 1) by HMEC-1 were measured in the supernatant and cell lysates, after incubation with 1 nM thrombin, fibrin with 3, and 30% fibrinogen γ/γ‘, using commercial kits. The influence of fibrinogen γ/γ‘ on fibrin structure on the surface of the HMEC-1 was followed with laser scanning confocal microscopy (LSCM). Results The kinetics of fibrin formation on HMEC-1 with 3 and 10% fibrinogen γ/γ‘ were similar. However, with 30% fibrinogen γ/γ‘ both the slope and final turbity were approximately 50% less. The LSCM images showed the dramatic effects of increasing fibrinogen γ/γ‘ from 3 to 30%. The uPA and PAI 1 concentrations in culture supernatants HMEC-1 cells treated with thrombin or 30% γ/γ‘ fibrin were two-fold increased as compared to basal culture supernatants and 3% γ/γ‘ fibrin-treated HMEC-1. In all stimulatory conditions the intracellular concentration of uPA was higher than in supernatants. In contrast, the intracellular PAI 1 concentration was decreased as compared to that measured in the supernatant, including the basal condition. Conclusion A concentration of 30% fibrin γ/γ‘ alter drastically fibrin structure on the cell surface and affects the secretion of uPA and PAI 1 through its capacity to bind thrombin.http://link.springer.com/article/10.1186/s12860-019-0233-0Fibrinogen gamma prime (γ’)Clot structureUrokinase plasminogen activator (uPA)Plasminogen activator inhibitor type 1 (PAI 1)Human dermal microvascular endothelial cells (HMEC-1) |
| spellingShingle | Miriam Cantero Héctor Rojas Eduardo Anglés-Cano Rita Marchi Fibrin γ/γ' influences the secretion of fibrinolytic components and clot structure BMC Molecular and Cell Biology Fibrinogen gamma prime (γ’) Clot structure Urokinase plasminogen activator (uPA) Plasminogen activator inhibitor type 1 (PAI 1) Human dermal microvascular endothelial cells (HMEC-1) |
| title | Fibrin γ/γ' influences the secretion of fibrinolytic components and clot structure |
| title_full | Fibrin γ/γ' influences the secretion of fibrinolytic components and clot structure |
| title_fullStr | Fibrin γ/γ' influences the secretion of fibrinolytic components and clot structure |
| title_full_unstemmed | Fibrin γ/γ' influences the secretion of fibrinolytic components and clot structure |
| title_short | Fibrin γ/γ' influences the secretion of fibrinolytic components and clot structure |
| title_sort | fibrin γ γ influences the secretion of fibrinolytic components and clot structure |
| topic | Fibrinogen gamma prime (γ’) Clot structure Urokinase plasminogen activator (uPA) Plasminogen activator inhibitor type 1 (PAI 1) Human dermal microvascular endothelial cells (HMEC-1) |
| url | http://link.springer.com/article/10.1186/s12860-019-0233-0 |
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