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Analysis of the influence of antithrombin on microvascular thrombosis: anti-inflammation is crucial for anticoagulation.

Sorg H, Hoffmann JO, Hoffmann JN, Vollmar B - Intensive Care Med Exp (2015)

Bottom Line: Experimental groups consisted of animals treated with AT or with tryptophan(49)-blocked AT (TrypAT), which exerts only anticoagulant but no anti-inflammatory effects.The antithrombotic capacity of AT significantly differs in the experimental groups in which anti-inflammation was antagonized.The anti-inflammatory influence of AT is essentially linked to its anticoagulant effect in the microvascular system.

View Article: PubMed Central - PubMed

Affiliation: Institute for Experimental Surgery, University Medicine Rostock, Schillingallee 69a, 18057, Rostock, Germany, heiko.sorg@krupp-krankenhaus.de.

ABSTRACT

Purpose: Microvascular thrombosis during septic conditions is of essential clinical relevance, but the pathomechanisms are not yet completely understood. The purpose of this study was to study the distinguished differentiation of the interactions of inflammation and coagulation using antithrombin (AT), a mediator of anticoagulation and anti-inflammation.

Methods: Using a thrombosis model in a cremaster muscle preparation of male C57Bl/6J mice (n = 83), we quantitatively assessed microvascular thrombus formation by using intravital fluorescence microscopy. Experimental groups consisted of animals treated with AT or with tryptophan(49)-blocked AT (TrypAT), which exerts only anticoagulant but no anti-inflammatory effects. To further see whether endothelial glycosaminoglycan (GAG) binding with consecutive prostacyclin (PGI2) release is mandatory for the anticoagulant process of AT, animals were administered heparin or indomethacin either alone or in combination with AT.

Results: The antithrombotic capacity of AT significantly differs in the experimental groups in which anti-inflammation was antagonized. This is given by the significantly prolonged occlusion times (p < 0.05) and higher patency rates in case of application of AT alone; while all other groups in which the anti-inflammatory action of AT was blocked by TrypAT, heparin or indomethacin revealed thrombus kinetics comparable to controls.

Conclusions: The anti-inflammatory influence of AT is essentially linked to its anticoagulant effect in the microvascular system. Those specifications of the active profile of AT characterize the intimate interactions of the anticoagulant and anti-inflammatory pathways. This might be of relevance for AT as a therapeutic agent in critically diseased patients and the clinical understanding of microvascular thrombosis.

No MeSH data available.


Related in: MedlinePlus

Complete vessel occlusion time. Time until complete occlusion of venules after induction of thrombus formation in cremaster muscle preparations of mice treated with either physiological saline (control), antithrombin (250 IU/kg; AT), tryptophan49-blocked AT (250 IU/kg; TrypAT), indomethacin (5 mg/kg; indo), indomethacin plus AT (indo + AT), heparin (100 IU/kg), or heparin plus AT (hep + AT). Data is given as means ± SEM; ANOVA, post-hoc comparison; * p < 0.01 vs. control; #p < 0.01 vs. AT
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Fig4: Complete vessel occlusion time. Time until complete occlusion of venules after induction of thrombus formation in cremaster muscle preparations of mice treated with either physiological saline (control), antithrombin (250 IU/kg; AT), tryptophan49-blocked AT (250 IU/kg; TrypAT), indomethacin (5 mg/kg; indo), indomethacin plus AT (indo + AT), heparin (100 IU/kg), or heparin plus AT (hep + AT). Data is given as means ± SEM; ANOVA, post-hoc comparison; * p < 0.01 vs. control; #p < 0.01 vs. AT

Mentions: CVO includes all clogged vessels within the investigation time of 1200 s and describes the mean time that was necessary for irreversible occlusion of a vessel by a thrombus (Fig. 4). In control animals, CVO was observed after 440 ± 54 s of ferric chloride and light exposure. AT application presented with major antithrombotic effectiveness, as given by a significant and more than 2-fold prolongation of the time (964 ± 69 s), which was needed for CVO (Fig. 4; p < 0.05 vs. control). To further analyze whether the antithrombotic effect is mandatorily linked to the binding of AT to GAGs, TrypAT has been used. Application of TrypAT showed significantly shorter occlusion times than AT (485 ± 79 s), which were comparable to that in controls, indicating that the GAG binding is being considered responsible for the observed anti-inflammatory capability of AT but is also necessary for its anticoagulant function. Additionally, we could observe that the combined application of indomethacin and AT (482 ± 57 s) as well as the indomethacin application alone (497 ± 108 s) could not prolong microvascular thrombus formation as given by similar values for CVO to those in controls. The combination of heparin and AT led to significantly prolonged CVO compared to the control group (790 ± 98 s, p < 0.05 vs. control); however, the effect is below the single AT application results.Fig. 4


Analysis of the influence of antithrombin on microvascular thrombosis: anti-inflammation is crucial for anticoagulation.

Sorg H, Hoffmann JO, Hoffmann JN, Vollmar B - Intensive Care Med Exp (2015)

Complete vessel occlusion time. Time until complete occlusion of venules after induction of thrombus formation in cremaster muscle preparations of mice treated with either physiological saline (control), antithrombin (250 IU/kg; AT), tryptophan49-blocked AT (250 IU/kg; TrypAT), indomethacin (5 mg/kg; indo), indomethacin plus AT (indo + AT), heparin (100 IU/kg), or heparin plus AT (hep + AT). Data is given as means ± SEM; ANOVA, post-hoc comparison; * p < 0.01 vs. control; #p < 0.01 vs. AT
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495092&req=5

Fig4: Complete vessel occlusion time. Time until complete occlusion of venules after induction of thrombus formation in cremaster muscle preparations of mice treated with either physiological saline (control), antithrombin (250 IU/kg; AT), tryptophan49-blocked AT (250 IU/kg; TrypAT), indomethacin (5 mg/kg; indo), indomethacin plus AT (indo + AT), heparin (100 IU/kg), or heparin plus AT (hep + AT). Data is given as means ± SEM; ANOVA, post-hoc comparison; * p < 0.01 vs. control; #p < 0.01 vs. AT
Mentions: CVO includes all clogged vessels within the investigation time of 1200 s and describes the mean time that was necessary for irreversible occlusion of a vessel by a thrombus (Fig. 4). In control animals, CVO was observed after 440 ± 54 s of ferric chloride and light exposure. AT application presented with major antithrombotic effectiveness, as given by a significant and more than 2-fold prolongation of the time (964 ± 69 s), which was needed for CVO (Fig. 4; p < 0.05 vs. control). To further analyze whether the antithrombotic effect is mandatorily linked to the binding of AT to GAGs, TrypAT has been used. Application of TrypAT showed significantly shorter occlusion times than AT (485 ± 79 s), which were comparable to that in controls, indicating that the GAG binding is being considered responsible for the observed anti-inflammatory capability of AT but is also necessary for its anticoagulant function. Additionally, we could observe that the combined application of indomethacin and AT (482 ± 57 s) as well as the indomethacin application alone (497 ± 108 s) could not prolong microvascular thrombus formation as given by similar values for CVO to those in controls. The combination of heparin and AT led to significantly prolonged CVO compared to the control group (790 ± 98 s, p < 0.05 vs. control); however, the effect is below the single AT application results.Fig. 4

Bottom Line: Experimental groups consisted of animals treated with AT or with tryptophan(49)-blocked AT (TrypAT), which exerts only anticoagulant but no anti-inflammatory effects.The antithrombotic capacity of AT significantly differs in the experimental groups in which anti-inflammation was antagonized.The anti-inflammatory influence of AT is essentially linked to its anticoagulant effect in the microvascular system.

View Article: PubMed Central - PubMed

Affiliation: Institute for Experimental Surgery, University Medicine Rostock, Schillingallee 69a, 18057, Rostock, Germany, heiko.sorg@krupp-krankenhaus.de.

ABSTRACT

Purpose: Microvascular thrombosis during septic conditions is of essential clinical relevance, but the pathomechanisms are not yet completely understood. The purpose of this study was to study the distinguished differentiation of the interactions of inflammation and coagulation using antithrombin (AT), a mediator of anticoagulation and anti-inflammation.

Methods: Using a thrombosis model in a cremaster muscle preparation of male C57Bl/6J mice (n = 83), we quantitatively assessed microvascular thrombus formation by using intravital fluorescence microscopy. Experimental groups consisted of animals treated with AT or with tryptophan(49)-blocked AT (TrypAT), which exerts only anticoagulant but no anti-inflammatory effects. To further see whether endothelial glycosaminoglycan (GAG) binding with consecutive prostacyclin (PGI2) release is mandatory for the anticoagulant process of AT, animals were administered heparin or indomethacin either alone or in combination with AT.

Results: The antithrombotic capacity of AT significantly differs in the experimental groups in which anti-inflammation was antagonized. This is given by the significantly prolonged occlusion times (p < 0.05) and higher patency rates in case of application of AT alone; while all other groups in which the anti-inflammatory action of AT was blocked by TrypAT, heparin or indomethacin revealed thrombus kinetics comparable to controls.

Conclusions: The anti-inflammatory influence of AT is essentially linked to its anticoagulant effect in the microvascular system. Those specifications of the active profile of AT characterize the intimate interactions of the anticoagulant and anti-inflammatory pathways. This might be of relevance for AT as a therapeutic agent in critically diseased patients and the clinical understanding of microvascular thrombosis.

No MeSH data available.


Related in: MedlinePlus