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- W2398207031 abstract "Wound healing is impaired in diabetes due to dysregulated inflammation. Previous studies by our group and others have demonstrated that inflammatory mediators are significantly increased in diabetic wounds; however, the mechanisms behind this hyperinflammatory response remain largely unknown. As there is known to be high levels of endotoxin in type 2 diabetic (T2D) wounds, we aimed to assess the role of high-fat diet (HFD) on Toll-like receptor 4 (TLR4) expression. We hypothesized that HFD/T2D would alter histone methylation, leading to increased receptor expression and enhanced inflammation. C57/BL6 mice were subjected to either normal or HFD for 12 to 16 weeks to induce the diet-induced obese (DIO) model of physiologic T2D. Four mm hind limb wounds were created and healing was monitored daily using NIH ImageJ software. Bone marrow-derived macrophages (BMDM) were cultured and TLR4 expression measured by qPCR. Myeloid TLR4 protein expression was assessed in peripheral blood and wound cell isolates by analytical flow cytometry. Wound macrophages (NK1.1−/Ly6G−/CD11b+) were isolated using magnetic sorting and inflammatory cytokine expression determined with qPCR and bioplex. Chromatin immunoprecipitation (ChIP) analysis at the NF-κB binding site on the TLR4 promoter was performed using antibodies to trimethylated histone 3 lysine 4 (H3K4me3; (n = ∼5 mice per group for all studies). DIO mice demonstrated significantly delayed wound healing as compared to littermate controls, and BMDM isolated from these mice revealed increased TLR4 gene expression. Correspondingly, peripheral blood and wound monocyte/macrophages showed increased TLR4 receptor levels by flow cytometry. ChIP analysis at the TLR4 promoter revealed significantly increased H3K4me3, a gene activating mark, in the HFD BMDM. Finally, macrophage wound isolates from DIO mice had elevated levels of inflammatory gene expression and cytokines. Enhanced H3K4me3 at the TLR4 promoter in diabetic macrophages increases receptor expression and may contribute to the hyperinflammatory response seen in T2D. Further mechanistic studies are needed in order to examine the role of TLR4 in wound healing." @default.
- W2398207031 created "2016-06-24" @default.
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- W2398207031 date "2016-06-01" @default.
- W2398207031 modified "2023-10-17" @default.
- W2398207031 title "PC238. Epigenetically Altered Toll-Like Receptor 4 Expression May Contribute to Increased Inflammation and Delayed Wound Healing in a Murine Model of Diabetes" @default.
- W2398207031 doi "https://doi.org/10.1016/j.jvs.2016.03.377" @default.
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