Recombinant human granulocyte colony-stimulating factor enhanced the resolution of venous thrombi

Abstract

Bone marrow-derived cells are recruited into the thrombus during resolution. This study explored whether mobilization of bone marrow cells with recombinant human granulocyte colony-stimulating factor (rhG-CSF) could enhance the resolution of venous thrombi and the accumulation of macrophages in thrombi and explored the effect of rhG-CSF on cysteine-cysteine chemokine receptor 2 (CCR2) expression. The Sprague-Dawley adult rats were randomly divided into four groups: control, sham-operated, thrombus, and treatment groups. Thrombi were induced in the thrombus and treatment group, which received a subcutaneous injection of rhG-CSF once daily for 6 days postoperatively. The thrombus, sham-operated, and control groups received equal volumes of 0.9% saline. The mononuclear cells in peripheral blood were analyzed by an automated hematology analyzer and counted under microscope. The cell marker CD68 was used to determine the number of macrophages in thrombi tissue sections. Levels of monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1alpha (MIP1alpha) in the peripheral blood were detected by enzyme-linked immunosorbent assay. Real-time reverse transcriptase-polymerase chain reaction and Western blot were used to analyze, respectively, the expression of CCR2 messenger RNA in the peripheral blood and CCR2 protein of THP-1 monocyte. At postoperative days 3 (P < .05) and 7 (P < .01), mononuclear cells significantly increased in treatment group (2.1 +/- 0.3, 4.4 +/- 0.3 x 10(6)/L) vs the thrombus group (1.7 +/- 0.2, 1.3 +/- 0.4 x 10(6)/L). The organization and recanalization of thrombi in treatment group progressed more quickly compared with the thrombus group (P < .01). The macrophage number of the thrombus in the treatment group (338 +/- 26 cells/15 high-power fields) increased significantly vs the thrombus group (125 +/- 11 cells/15 high-power fields, P < .01). No statistical difference was observed between the thrombus and treatment group in the MCP-1 and MIP-1alpha level in peripheral blood. Expressions of the CCR2 gene in the peripheral blood of the treatment group significantly increased compared with the thrombus group (P < .05). Recombinant human G-CSF induced higher expression of CCR2 protein of human monocytic cell line THP-1. Bone marrow mobilization enhanced the resolution and recanalization of venous thrombi. This process was associated with increased macrophage accumulation in thrombi, which might be the result of higher CCR2 expression of monocytes. The classic treatment of venous thrombi is anticoagulation. Anticoagulant therapy and thrombolysis both have limited effects on existing thrombi and have a small but significant risk of severe hemorrhage. In clinical practice, we lack specific treatment for patients with venous thrombosis combined with brain hemorrhage or a gastrointestinal activated ulcer, which are contraindicated for anticoagulation and thrombolytic therapy. Enhancing the resolution of venous thrombi would contribute to its therapy. Bone marrow-derived cells are recruited into the thrombus during resolution. Many of these cells express a macrophage phenotype and may represent a population of plastic stem cells that orchestrate thrombus recanalization. Recombinant human granulocyte colony stimulating factor (rhG-CSF) can mobilize monocytic lineage cells into peripheral blood and may contribute to this cell in the thrombi. If rhG-CSF enhances the resolution of venous thrombi and recanalization, it might be used to treat patients with venous thrombi, especially those who have contraindication for anticoagulation and thrombolytic therapy.