Effects Of Recombinant Human Interleukin-1β On Functions Of Peripheral Blood Mononuclear Cells From Healthy Adults And The Role Of Paeoniflorin

OBJECTIVETo investigate the effects of paeoniflorin (Pae) on peripheral blood mononuclear cell (PBMC) of healthy adults. we used recombinant human interleukin-1β(rhIL-1β) stimulating PBMC from healthy volunteers in vitro, and observed the effects of Pae on phagocytosis function, tumor necrosis factor-α(TNF-α) and prostaglandin E₂ (PGE₂) production, expression of human leukocyte antigen-DR (HLA-DR) and CD80 of activated PBMC, in order to provide evidence for further elucidating the mechanisms of Pae. METHODSAfter informed consent, collected peripheral venous blood of healthy volunteer under the condition of asepsis and empty stomach in morning. Applied Ficoll density gradient centrifugation and adherence method to obtain PBMC, and PBMC was incubated with rhIL-1β(0.01, 0.1, 1, 10μg·L^-1) for 24 hours or rhIL-1β(1μg·L^-1) for 3, 6, 12, 24 hours, then Pae (10-9, 10-8, 10-7, 10-6, 10-5mol·L^-1) was added to culture 24 hours. Then cultured cells and supernatant fluids were collected. Phagocytosis function was measured by neutral red staining method; and the levels of TNF-αand PGE₂ were detected by radioimmunoassay (RIA) and enzyme linked immunosorbent assay (ELISA) respectively, also the mean fluorescence rates of HLA-DR and CD80 expressed on PBMC were detected by flow cytometry analysis.RESULTS1. The effects of rhIL-1βon elevating phagocytosis function and TNF-αand PGE₂ level of PBMC from healthy adults in vitroIn the research, we separated PBMC from healthy adults, and chose rhIL-1βas stimulating factor, to observe the effects of different concentration of rhIL-1β(0.01, 0.1, 1, 10μg·L^-1) and rhIL-1β(1μg·L^-1) cultured different time on phagocytosis function and TNF-αand PGE₂ level of PBMC. The results suggested that rhIL-1βcould elevate phagocytosis function and TNF-αand PGE₂ level of PBMC .2. The effects of rhIL-1βon elevating the expression of HLA-DR and CD80 on PBMC from healthy adults in vitroIn vitro experiment, we observed the effects of different concentration of rhIL-1β(0.01, 0.1, 1, 10μg·L^-1) and rhIL-1β(1μg·L^-1) cultured different time on expression of HLA-DR and CD80 of PBMC. rhIL-1βcould promote the mean fluorescence rate of HLA-DR and CD80 of PBMC, and the mean fluorescence rate of HLA-DR increased as cultured time extended, presenting time dependence (r=0.977, P<0.05). 3. The effects of Pae on inhibiting phagocytosis function and TNF-αand PGE₂ level of PBMC stimulated by rhIL-1βin vitroThe administration of Pae (10^-9,10^-8,10^-7,10^-6,10^-5mol·L^-1) in vitro inhibited phagocytosis function, and decreased the level of TNF-αand PGE₂ of PBMC by rhIL-1β(1μg·L^-1) as concentration increased, presenting concentration dependence (TNF-α, r=-0.820, P<0.05 and PGE₂, r=-0.836, P<0.05).4. The effects of Pae on inhibiting the expression of HLA-DR and CD80 of PBMC stimulated by rhIL-1βin vitroThe results in vitro experiment, Pae(10^-9,10^-8,10^-7,10^-6,10^-5mol·L^-1) suppressed the expression of HLA-DR and CD80 of PBMC by rhIL-1β(1μg·L^-1), decreasing the mean fluorescence rate of HLA-DR and CD80 of PBMC .CONCLUSIONS1. IL-lβcould promote PBMC activation, enhancing phagocytosis function, TNF-αand PGE₂ production, also expression of HLA-DR and CD80.2. Pae could inhibit PBMC activation stimulated by IL-lβ, decreasing phagocytosis function, TNF-αand PGE₂ production, also expression of HLA-DR and CD80.