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

Background: 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. Methods: 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-1 alpha (MIP1 alpha) 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. Results: 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-1 alpha 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. Conclusions: 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.