ABSTRACT
【Objective】 To investigate the effects of naringenin on the polarization of high-glycemic RAW264.7 macrophages and its related mechanism. 【Methods】 The mother solution of NAR was prepared with dimethyl sulfoxide (DMSO), and the log-growth phase macrophage RAW264.7 was pre-tested. DMEM medium with different glucose concentrations (1, 2, 4, 5 and 6 g/L) was used for cultivation for 24 h. Before the experiment, DMEM was diluted into NAR mixture with different final concentrations, and the effect of NAR on RAW264.7 cell activity was detected by CCK-8 method; nitric oxide synthase (NOS) type classification determination of checkerboard induced nitric oxide synthase (iNOS) active filter was used to control the concentration of sugar and high-sugar stimulation. The control group were subdivided into normal control (NG) and osmotic pressure control (NG+M). The high-glucose stimulation group was divided into normal high glucose (HG), high glucose + naringenin (HG+NAR), high glucose + Fasudil (HG+F), and high glucose +C3 transferase (HG+C3). RAW264.7 was cultured for 24 h in each group; the expression levels of supernatant cytokines, namely, interleukin6 (il-6), tumor necrosis factor -α (TNF-α) and interleukin10 (IL-10), were detected by ELISA. Western blotting was used to determine the RhoA/ROCK pathway related proteins, iNOS and Arg-1 protein levels. Type (M1, M2) and proportion (M1/M2) of macrophages were analyzed by flow cytometry. 【Results】 Compared with those in NG group, in HG group RhoA/ROCK pathway-related proteins and iNOS expression were increased, while Arg-1 expression was decreased (P<0.05). The secretion of pro-inflammatory cytokines IL-6 and TNF-α was increased while anti-inflammatory cytokine IL-10 was decreased (P<0.05). The number of M1-type cells and M1/M2 ratio increased (P<0.05). Compared with HG group, RhoA/ROCK pathway related proteins and iNOS expression were decreased in HG+NAR group, HG+F group and HG+C3 group, while Arg-1 expression was increased, IL-6 and TNF- α secretion was decreased, IL-10 was increased, M2-type macrophages were increased, and M1/M2 was decreased (P<0.05). 【Conclusion】 NAR may promote the M2-type differentiation of macrophages stimulated by high glucose by down-regulating RhoA/ROCK signaling pathway.
ABSTRACT
This study was aimed to investigate the changes of cytokine contents in single donor platelets (SDPs) collected by using MCS(+), Trima, Amicus blood cell separators during storage. 18 portions of SDPs were collected by MCS(+), Trima, Amicus blood cell separators, were preserved in standard condition of blood bank, the levels of cytokines such as IL-8, RANTES, CD154, TGF-beta(1) and VEGF were detected by ELISA at 1, 3, 5, 7 days during storage. The results showed that the levels of IL-8, RANTES, CD154, TGF-beta(1) and VEGF in SDPs collected by blood cell separators MCS(+), Trima, and Amicus gradually increased with prolonging of time during storage, but the increase of IL-8 level in SDPs collected by MCS(+) separator was significant difference from SDPs collected by Trime and Amicus separators (p < 0.05). It is concluded that the all collected SDPs mentioned above express IL-8, RANTES, CD154, TGF-beta(1) and VEGF during storage, and their cytokine levels show a tendency to increase with prolonging of time during storage, apheresis platelets with less leukocytes express IL-8 lower.
Subject(s)
Humans , Blood Platelets , Metabolism , CD40 Ligand , Blood , Cell Separation , Methods , Chemokine CCL5 , Blood , Cytokines , Blood , Interleukin-8 , Blood , Platelet Count , Specimen Handling , Transforming Growth Factor beta , Blood , Vascular Endothelial Growth Factor A , BloodABSTRACT
To evaluate flow-cytometry and Nageotte method for counting residual WBC in thrombocytaphoresis concentrates, their accuracies were determined by dilution studies separately; the repeatability was determined by measuring the interassay coefficient of variation for 14 replicates of a sample with known leukocyte concentration. 102 samples of leukocyte-depleted thrombocytaphoresis concentrates were detected by the above mentioned two methods, and the results were compared each other. The results showed that no difference was observed between two methods over a range of leukocyte concentrations from 0.8 to 10 WBC/microl (P > 0.05). In conclusion, flow-cytometry and Nageotte methods can be used for quality control of WBC-reduced blood components.