Preliminary study of stored red blood cell metabolites persistently affecting macrophage function
Objective:To investigate the effect of metabolites of stored red blood cells(RBCs)on the function of macrophages.Methods:The macrophages were isolated and cultured from human peripheral blood,then CD14 and CD68 macrophages were detected by flow cytometry.The contents of IL-4 and IL-6 in the supernatant which the RBCs stored for 21 days and co-cultured with the macrophages were detected,and the expression of CD68 pro-tein in macrophages was detected by immunofluorescence method at 0 h,12 h,24 h and 36 h,respectively.Results:Suitable macrophages could be cultured by both adherence method and magnetic beads method,but the purity of macrophages obtained by magnetic beads was higher.The expression of CD68 protein in macrophages was different after different time of co-culture with RBCs,and the expression of CD68 was the highest in the 24-hour group.The secretion of IL-4 and IL-6 in supernant was also different,and the content of IL-4 was 31.21 pg/mL,31.55 pg/mL,31.77 pg/mL and 31.73 pg/mL at 0 h,12 h,24 h and 36 h,respectively,with no statistical difference(P＞0.05).The content of IL-6 was 2.34 pg/mL,25.56 pg/mL,65.15 pg/mL and 35.47 pg/mL at 0 h,12 h,24 h and 36 h,respectively,with significant differences(P＜0.05).Conclusion:The reliability of the method for isolating and culturing macrophages in peripheral blood of humans was high.The study on the func-tional changes of macrophages stimulated by stored red blood cells and inflammatory factors found that metabolic products of stored red blood cells had pro-inflammatory effects on the internal environment of the body,which could provide theoretical references and data support for further research on the impact of stored red blood cells on macrophage function.
stored red blood cellshuman peripheral bloodmacrophagesinflammatory factors
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