[Temporal Expression of NETosis Marker CitH3 in Deep Vein Thrombosis in Mice].

Objectives: To detect the expression changes of citrullinated histone H3 (CitH3) during the development of deep vein thrombosis (DVT) in mice, and to explore its value in estimating the time to thrombosis.

Methods: The inferior vena cava (IVC) of mice was ligated to establish a thrombosis model induced by congestion. Mice were sacrificed under excessive anesthesia at 0 h, 1 d, 3 d, 5 d, 7 d, 10 d, 14 d and 21 d after the modeling, respectively. The congested IVC segments (0 h after modeling) and the thrombosed IVC segments (1-21 days after modeling) were extracted. Immunohistochemistry and double immunofluorescence staining were used to observe the number of neutrophils and the expression of CitH3 during thrombosis. Western blotting was used to detect the protein expression level of CitH3.

Results: During thrombosis, CitH3 was mainly expressed in neutrophils within the thrombus. A small number of neutrophils and a few CitH3-positive cells were observed at 0 h after modeling in the congested IVC. Between 1 d and 21 d after modeling, the number of neutrophils reached a peak at 1 d and gradually decreased. The number of CitH3-positive cells and their ratio to neutrophils began to increase at 1 d, reached a peak at 5 d after modeling, and then decreased. The expression level of CitH3 protein began to increase at 1 d and reached a peak at 5 d after modeling.

Conclusions: The expression of CitH3 during DVI shows temporal changes, and is expected to become a biological marker for estimating the formation time of thrombosis.