Abstract:

Objective To evaluate the effects of different anticoagulants on phagocytosis，activation and apoptosis of in vitro neutrophil function. Methods Totally，30 healthy subjects who underwent physical examination in Henan Provincial People's Hospital from January 2020 to June 2020 were enrolled，and they were classified into 3 groups randomly（10 cases for each group）. The 3 mL peripheral blood was collected with heparin sodium anticoagulant tube（heparin sodium group），2 mL peripheral blood was collected with ethylenediaminetetraacetic acid（EDTA）-K₂ anticoagulant tube（EDTA-K₂ group），and 2 mL peripheral blood was collected with sodium citrate anticoagulant tube（sodium citrate group）. The average fluorescence intensity of OVA-FITC was determined by flow cytometry to determine the phagocytosis ability of neutrophils. The expressions of neutrophil surface activation markers，CD62L and CD11B，were determined to assess the activation of neutrophils. 7AAD and Annexin-V were added to determine the apoptosis of neutrophils. Results The average fluorescence intensity of OVA-FITC phagocytosis neutrophils in EDTA-K₂ group was lower than those in heparin sodium and sodium citrate groups（P<0.05）. CD11B neutrophils in EDTA-K₂ group were lower than those in heparin sodium and sodium citrate groups（P<0.01，P<0.05）. Compared with heparin sodium group，neutrophil apoptosis was increased in EDTA-K₂ and sodium citrate groups（P<0.01，P<0.05）. Conclusions Different anticoagulants can affect the phagocytosis，activation and apoptosis of neutrophils in vitro，but each anticoagulant has different effects. Therefore，appropriate anticoagulant must be selected to accurately determine the phagocytosis，activation and apoptosis of neutrophils.

Key words: Neutrophil, Phagocytosis, Activation, Apoptotic function, Anticoagulant