High-Dose Tranexamic Acid Enhances Circulating Neutrophil Extracellular Traps and Thrombus in Thrombosis Mouse Model

<b>Background/Objectives</b>: Tranexamic acid (TXA) reduces mortality in patients with massive hemorrhage by inhibiting fibrinolysis. However, it is associated with an increased risk of thrombosis. The activation of neutrophil extracellular traps (NETs) has been implicated in the formati...

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Main Authors: Jung-Wook Song, Eun-Hye Seo, Un Yung Choi, Chung-Sik Oh, Aram Kim, Keeho Song, Seung-Hyun Lee, Jin Kook Kim
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Language:English
Published: MDPI AG 2025-05-01
Series:Biomedicines
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Online Access:https://www.mdpi.com/2227-9059/13/6/1284
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author Jung-Wook Song
Eun-Hye Seo
Un Yung Choi
Chung-Sik Oh
Aram Kim
Keeho Song
Seung-Hyun Lee
Jin Kook Kim
author_facet Jung-Wook Song
Eun-Hye Seo
Un Yung Choi
Chung-Sik Oh
Aram Kim
Keeho Song
Seung-Hyun Lee
Jin Kook Kim
author_sort Jung-Wook Song
collection DOAJ
description <b>Background/Objectives</b>: Tranexamic acid (TXA) reduces mortality in patients with massive hemorrhage by inhibiting fibrinolysis. However, it is associated with an increased risk of thrombosis. The activation of neutrophil extracellular traps (NETs) has been implicated in the formation of thrombosis. This study investigated the effects of tranexamic acid on circulating and localized NETs, neutrophils, platelets, and the vascular endothelium in a mouse model of thrombosis. <b>Methods</b>: A ferric chloride-induced thrombosis mouse model was used and divided into five groups: a Control group that received intraperitoneal phosphate-buffered saline (PBS), and four experimental groups that received intraperitoneal tranexamic acid at doses of 5 mg/kg, 10 mg/kg, 20 mg/kg, and 30 mg/kg, respectively. To evaluate the expression of circulating and localized NETs, neutrophils, platelets, vascular endothelial cells, fibrinogen, and D-dimer, the following markers were analyzed: myeloperoxidase (MPO), neutrophil marker, cluster of differentiation (CD)31, CD34, fibrinogen α-chain, and D-dimer. These markers were assessed using flow cytometry, immunohistofluorescence staining, and Western blot analysis. The primary endpoint was the differential expression of anti-MPO antibody among the groups. <b>Results</b>: In total, data from 20 thrombosis mouse models were analyzed. For each group, four samples were assessed by flow cytometry, and three samples by immunohistofluorescence staining and Western blot analysis, respectively. In the flow cytometric analysis, circulating anti-MPO antibody expression was significantly higher in the TXA 20 and TXA 30 groups compared to the Control group (<i>p</i> = 0.001 and <i>p</i> = 0.001, respectively). Immunohistofluorescence staining revealed that D-dimer expression in the thrombotic femoral artery was significantly lower in the TXA 5, TXA 10, and TXA 20 groups compared to the Control group (<i>p</i> = 0.005; <i>p</i> = 0.018; <i>p</i> = 0.004, respectively), but significantly higher in the TXA 30 group than in the Control group (<i>p</i> = 0.044). Similarly, the expression of anti-fibrinogen antibody was significantly lower in the TXA 5, TXA 10, and TXA 20 groups compared to the Control group (<i>p</i> = 0.038; <i>p</i> = 0.003; <i>p</i> = 0.041, respectively). Western blot analysis showed no significant differences in the expression of anti-Ly6B.2, anti-fibrinogen, and anti-CD31 antibodies among the groups. <b>Conclusions</b>: The present study suggests that high-dose tranexamic acid (30 mg/kg) administration may increase circulating NETs and localized D-dimer levels, indicating a higher potential for thrombosis in a thrombosis mouse model. These findings imply that the prothrombotic effects of tranexamic acid may be dose-dependent and could vary based on underlying disease conditions. Therefore, the careful dosage adjustment of tranexamic acid may be necessary, particularly in patients at risk of thrombosis.
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spelling doaj-art-1c17fa686a9a417285ae55e80b32dd732025-08-20T03:32:32ZengMDPI AGBiomedicines2227-90592025-05-01136128410.3390/biomedicines13061284High-Dose Tranexamic Acid Enhances Circulating Neutrophil Extracellular Traps and Thrombus in Thrombosis Mouse ModelJung-Wook Song0Eun-Hye Seo1Un Yung Choi2Chung-Sik Oh3Aram Kim4Keeho Song5Seung-Hyun Lee6Jin Kook Kim7Department of Infection and Immunology, Konkuk University School of Medicine, Seoul 05030, Republic of KoreaKorea mRNA Vaccine Initiative, Gachon University, Seongnam 13120, Republic of KoreaDepartment of Microbiology, Konkuk University School of Medicine, Chungju 27478, Republic of KoreaDepartment of Infection and Immunology, Konkuk University School of Medicine, Seoul 05030, Republic of KoreaDepartment of Urology, KonKuk University Medical Center, KonKuk University School of Medicine, Seoul 05030, Republic of KoreaDivision of Endocrinology and Metabolism, Department of Internal Medicine, Konkuk University Medical Center, Konkuk University School of Medicine, Seoul 05030, Republic of KoreaDepartment of Infection and Immunology, Konkuk University School of Medicine, Seoul 05030, Republic of KoreaDepartment of Otorhinolaryngology-Head and Neck Surgery, Konkuk University Medical Center, Konkuk University School of Medicine, Seoul 05030, Republic of Korea<b>Background/Objectives</b>: Tranexamic acid (TXA) reduces mortality in patients with massive hemorrhage by inhibiting fibrinolysis. However, it is associated with an increased risk of thrombosis. The activation of neutrophil extracellular traps (NETs) has been implicated in the formation of thrombosis. This study investigated the effects of tranexamic acid on circulating and localized NETs, neutrophils, platelets, and the vascular endothelium in a mouse model of thrombosis. <b>Methods</b>: A ferric chloride-induced thrombosis mouse model was used and divided into five groups: a Control group that received intraperitoneal phosphate-buffered saline (PBS), and four experimental groups that received intraperitoneal tranexamic acid at doses of 5 mg/kg, 10 mg/kg, 20 mg/kg, and 30 mg/kg, respectively. To evaluate the expression of circulating and localized NETs, neutrophils, platelets, vascular endothelial cells, fibrinogen, and D-dimer, the following markers were analyzed: myeloperoxidase (MPO), neutrophil marker, cluster of differentiation (CD)31, CD34, fibrinogen α-chain, and D-dimer. These markers were assessed using flow cytometry, immunohistofluorescence staining, and Western blot analysis. The primary endpoint was the differential expression of anti-MPO antibody among the groups. <b>Results</b>: In total, data from 20 thrombosis mouse models were analyzed. For each group, four samples were assessed by flow cytometry, and three samples by immunohistofluorescence staining and Western blot analysis, respectively. In the flow cytometric analysis, circulating anti-MPO antibody expression was significantly higher in the TXA 20 and TXA 30 groups compared to the Control group (<i>p</i> = 0.001 and <i>p</i> = 0.001, respectively). Immunohistofluorescence staining revealed that D-dimer expression in the thrombotic femoral artery was significantly lower in the TXA 5, TXA 10, and TXA 20 groups compared to the Control group (<i>p</i> = 0.005; <i>p</i> = 0.018; <i>p</i> = 0.004, respectively), but significantly higher in the TXA 30 group than in the Control group (<i>p</i> = 0.044). Similarly, the expression of anti-fibrinogen antibody was significantly lower in the TXA 5, TXA 10, and TXA 20 groups compared to the Control group (<i>p</i> = 0.038; <i>p</i> = 0.003; <i>p</i> = 0.041, respectively). Western blot analysis showed no significant differences in the expression of anti-Ly6B.2, anti-fibrinogen, and anti-CD31 antibodies among the groups. <b>Conclusions</b>: The present study suggests that high-dose tranexamic acid (30 mg/kg) administration may increase circulating NETs and localized D-dimer levels, indicating a higher potential for thrombosis in a thrombosis mouse model. These findings imply that the prothrombotic effects of tranexamic acid may be dose-dependent and could vary based on underlying disease conditions. Therefore, the careful dosage adjustment of tranexamic acid may be necessary, particularly in patients at risk of thrombosis.https://www.mdpi.com/2227-9059/13/6/1284thrombosistranexamic acidneutrophilplateletendothelium
spellingShingle Jung-Wook Song
Eun-Hye Seo
Un Yung Choi
Chung-Sik Oh
Aram Kim
Keeho Song
Seung-Hyun Lee
Jin Kook Kim
High-Dose Tranexamic Acid Enhances Circulating Neutrophil Extracellular Traps and Thrombus in Thrombosis Mouse Model
Biomedicines
thrombosis
tranexamic acid
neutrophil
platelet
endothelium
title High-Dose Tranexamic Acid Enhances Circulating Neutrophil Extracellular Traps and Thrombus in Thrombosis Mouse Model
title_full High-Dose Tranexamic Acid Enhances Circulating Neutrophil Extracellular Traps and Thrombus in Thrombosis Mouse Model
title_fullStr High-Dose Tranexamic Acid Enhances Circulating Neutrophil Extracellular Traps and Thrombus in Thrombosis Mouse Model
title_full_unstemmed High-Dose Tranexamic Acid Enhances Circulating Neutrophil Extracellular Traps and Thrombus in Thrombosis Mouse Model
title_short High-Dose Tranexamic Acid Enhances Circulating Neutrophil Extracellular Traps and Thrombus in Thrombosis Mouse Model
title_sort high dose tranexamic acid enhances circulating neutrophil extracellular traps and thrombus in thrombosis mouse model
topic thrombosis
tranexamic acid
neutrophil
platelet
endothelium
url https://www.mdpi.com/2227-9059/13/6/1284
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