Research Article

Oxygenator Is the Main Responsible for Leukocyte Activation in Experimental Model of Extracorporeal Circulation: A Cautionary Tale

Figure 1

P38 and NF-κB activation in leukocytes in response to ECC was significantly higher in presence of oxygenator. Peripheral blood samples were collected at baseline, before and after ECC. Leukocytes were fixed and permeabilized before intracellular staining using Alexa Fluor 568-conjugated antibodies that recognize (a) Thr180/Tyr182 phosphorylated p38 or (b) Ser529 phosphorylated NF-κB or with isotype-matched antibodies as a control. After lysis of red blood cells, fluorescence of granulocytes or mononuclear cells was quantified by flow-cytometry (after gating of cells by size and granularity). Mean fluorescence levels were calculated after subtracting values from isotype-control antibodies. Mean values are shown with standard deviations. between the two groups at the same time point.