Fig. 4. Paeonol improves the repair of high glucose-induced cell damage by modulating macrophage polarization.
(A) MTS kit was used to assess the effect of paeonol treatment (0, 5, 10, 25, and 50 μM) on macrophage viability. RAW264.7 cells were polarized into M1 and M2 macrophages under LPS + IFN-γ treatment and IL-4 treatment, respectively. Then M1 or M2 macrophages were treated with paeonol and RT-qPCR (B) and immunofluorescence (C) was used to detect the expression of the M1 marker iNOS and the M2 marker arginase 1. Next, HG-treated mouse skin fibroblasts were co-cultured with M1 or M2-polarized macrophages treated with or without paeonol. (D, E) Western blot was used to detect the expression of inflammatory factors (IL-1β, TNF-α, IL-4, and IL-10), angiogenic factors (CD31 and VEGFA), and collagen I/III in the supernatant of mouse skin fibroblasts. Scale bar = 50 μm. The data were presented as mean ± standard deviation of three independent replicate experiments. *p < 0.05, compared with the control group. *p < 0.05, **p < 0.01, ***p < 0.001, compared with the control group. #p < 0.05, compared with the HG, LPS + IFN-γ, or LPS + IL-4 group. &p < 0.05, &&p < 0.01, &&&p < 0.001, compared with the HG + M1 or HG + M2 group. HG, high glucose; LPS, lipopolysaccharide; IFN, interferon; IL, interleukin; TNF-α, tumor necrosis factor α; RT-qPCR, Reverse transcription-quantitative polymerase chain reaction.
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