Volume 45, Issue 8 e70223

ORIGINAL ARTICLE

Platelet-Induced NET in Liver Cirrhosis: Mitochondrial ROS-Mediated NETosis and Its Contribution to the Hypercoagulable State

Xiaoming Wu,

Xiaoming Wu

orcid.org/0000-0002-4271-8534

Department of Hematology, The First Hospital, Harbin Medical University, Harbin, China

The Key Laboratory of Myocardial Ischemia, Ministry of Education, Harbin, China

School of Basic Medical Sciences, Harbin Medical University, Harbin, China

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Haijiao Jing,

Haijiao Jing

orcid.org/0000-0002-8506-4016

Department of Hematology, The First Hospital, Harbin Medical University, Harbin, China

The Key Laboratory of Myocardial Ischemia, Ministry of Education, Harbin, China

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Tao Jiang,

Tao Jiang

Department of General Surgery, The First Hospital, Harbin Medical University, Harbin, China

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Cong Zhang,

Cong Zhang

Department of Hematology, The First Hospital, Harbin Medical University, Harbin, China

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Mengdi Li,

Mengdi Li

Department of Hematology, The First Hospital, Harbin Medical University, Harbin, China

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Huan Liu,

Huan Liu

Department of Hematology, The First Hospital, Harbin Medical University, Harbin, China

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Valerie A. Novakovic,

Valerie A. Novakovic

Department of Research, VA Boston Healthcare System, Harvard Medical School, Boston, Massachusetts, USA

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Shaohong Fang,

Shaohong Fang

The Key Laboratory of Myocardial Ischemia, Ministry of Education, Harbin, China

Department of Cardiology, Second Affiliated Hospital of Harbin Medical University, Harbin, China

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Jialan Shi,

Corresponding Author

Jialan Shi

orcid.org/0000-0001-5642-5436

Department of Hematology, The First Hospital, Harbin Medical University, Harbin, China

Department of Research, VA Boston Healthcare System, Harvard Medical School, Boston, Massachusetts, USA

Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts, USA

Correspondence:

Jialan Shi ([email protected]; [email protected])

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Xiaoming Wu,

Xiaoming Wu

orcid.org/0000-0002-4271-8534

Department of Hematology, The First Hospital, Harbin Medical University, Harbin, China

The Key Laboratory of Myocardial Ischemia, Ministry of Education, Harbin, China

School of Basic Medical Sciences, Harbin Medical University, Harbin, China

Search for more papers by this author

Haijiao Jing,

Haijiao Jing

orcid.org/0000-0002-8506-4016

Department of Hematology, The First Hospital, Harbin Medical University, Harbin, China

The Key Laboratory of Myocardial Ischemia, Ministry of Education, Harbin, China

Search for more papers by this author

Tao Jiang,

Tao Jiang

Department of General Surgery, The First Hospital, Harbin Medical University, Harbin, China

Search for more papers by this author

Cong Zhang,

Cong Zhang

Department of Hematology, The First Hospital, Harbin Medical University, Harbin, China

Search for more papers by this author

Mengdi Li,

Mengdi Li

Department of Hematology, The First Hospital, Harbin Medical University, Harbin, China

Search for more papers by this author

Huan Liu,

Huan Liu

Department of Hematology, The First Hospital, Harbin Medical University, Harbin, China

Search for more papers by this author

Valerie A. Novakovic,

Valerie A. Novakovic

Department of Research, VA Boston Healthcare System, Harvard Medical School, Boston, Massachusetts, USA

Search for more papers by this author

Shaohong Fang,

Shaohong Fang

The Key Laboratory of Myocardial Ischemia, Ministry of Education, Harbin, China

Department of Cardiology, Second Affiliated Hospital of Harbin Medical University, Harbin, China

Search for more papers by this author

Jialan Shi,

Corresponding Author

Jialan Shi

orcid.org/0000-0001-5642-5436

Department of Hematology, The First Hospital, Harbin Medical University, Harbin, China

Department of Research, VA Boston Healthcare System, Harvard Medical School, Boston, Massachusetts, USA

Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts, USA

Correspondence:

Jialan Shi ([email protected]; [email protected])

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First published: 12 July 2025

https://doi.org/10.1111/liv.70223

Handling Editor: Luca Valenti

Funding: This work was supported by The Postdoctoral Fellowship Program of China Postdoctoral Science Foundation (GZC20240352).

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ABSTRACT

Background & Objectives

Relatively little is known about mitochondrial ROS (mtROS)-mediated neutrophil extracellular traps (NETs) release. The aim of this study was to investigate whether the elevated NETs in liver cirrhosis were induced by platelets via mtROS and their effects on the cirrhotic coagulation system.

Methods

Patients with cirrhosis (n = 103) and healthy controls (n = 19) were included in the study. Platelet-induced NETosis was measured by immunofluorescence staining, confocal microscopy, flow cytometry, and microplate assays. NETs’ procoagulant activity was assessed using purified coagulation complex assays and thrombin formation in cirrhotic plasma.

Results

NETs’ levels were elevated in Child-Pugh B and C patients. Their platelets or high mobility group box 1 (HMGB1) increased NETs release of autologous neutrophils. Cirrhotic neutrophils had an increase in mtROS levels, which were enhanced by autologous platelets. Importantly, Mito TEMPO (a mitochondrial ROS inhibitor) inhibited platelet-induced NETs’ formation. Higher levels of HMGB1 on platelets and neutrophil autophagy were detected in Child-Pugh B and C patients. Their platelets or HMGB1 increased autophagy levels of autologous neutrophils. Furthermore, anti-HMGB1, anti-RAGE (receptor for advanced glycation endproducts) antibodies, and wortmannin inhibited platelet-induced autophagy and NETs formation. Subsequently, we found some differences between platelet- and PMA-induced NETosis, and further studied the dynamic changes of platelet-mediated NETosis. Lastly, these elevated NETs increased FXa, thrombin, and fibrin formation, shortened coagulation time, decreased thrombomodulin levels on endothelial cells, and impaired thrombomodulin activity.

Conclusions

Cirrhotic platelets induced NETs formation through mtROS and autophagy, which heightened the procoagulant activity and impaired the anticoagulant activity of thrombomodulin.

Conflicts of Interest

The authors declare no conflicts of interest.

Open Research

Data Availability Statement

Data available on request from authors.

Supporting Information

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Volume45, Issue8

August 2025

e70223

Platelet-Induced NET in Liver Cirrhosis: Mitochondrial ROS-Mediated NETosis and Its Contribution to the Hypercoagulable State

ABSTRACT

Background & Objectives

Methods

Results

Conclusions

Conflicts of Interest

Open Research

Data Availability Statement

Supporting Information

References

References

Information

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Platelet-Induced NET in Liver Cirrhosis: Mitochondrial ROS-Mediated NETosis and Its Contribution to the Hypercoagulable State

ABSTRACT

Background & Objectives

Methods

Results

Conclusions

Conflicts of Interest

Open Research

Data Availability Statement

Supporting Information

References

References

Related

Information