Journal: Cell Reports Medicine

Article Title: Enalaprilat reverses neutrophil polarization imbalance via targeting taurine-STING axis for treatment of diabetic wounds

doi: 10.1016/j.xcrm.2026.102714

Figure Lengend Snippet: Phenotype repolarization of neutrophils induced by Ena affected the pro-angiogenic capacities of HUVECs and inflammatory state of macrophages (A) Flow cytometry was adopted to reveal the difference of CD206+ population ratio among the three groups. n = 3 independent experiments. (B) Effects of AGE stimulation and sequential Ena supplement on the expression of ICAM1, CXCR2, CXCR4, and Fas as evaluated by western blot. (C) Effects of AGE stimulation and sequential Ena supplement on the expression of ICAM1, CXCR2, CXCR4, and Fas as evaluated by immunofluorescence staining. Scale bar, 2 μm; n = 3 independent experiments. (D) Quantitative reverse-transcription PCR (RT-qPCR) was employed to quantify the level of il1b , ccl3 , arg1 , and ccl17 . n = 3 independent experiments. (E) Schematic diagram for the incubation of HUVECs with pretreated neutrophils. (F) Proliferative ability of HUVECs under the stimulation of neutrophils with indicated preconditioning as detected via EdU staining. Scale bar, 200 μm; n = 3 independent experiments. (G) Migration property of HUVECs as measured by transwell assay. Scale bar, 200 μm; n = 3 independent experiments. (H) Migration property of HUVECs as measured by wound scratch test. Scale bar, 500 μm. (I) Tube formation assay was performed to visualize the neovascularization function of HUVECs with different treatments. Scale bar, 200 μm; n = 3 independent experiments. (J) Phenotype characters of macrophages irritated by neutrophils as determined by immunofluorescence staining. Scale bar, 5 μm. (K) Phenotype characters of macrophages irritated by neutrophils as determined by flow cytometry. n = 3 independent experiments. Data were shown as mean ± standard deviation (SD) from biological replicates, and statistical analyses were performed using one-way ANOVA test followed by Tukey’s multiple comparisons test in (A–D and F–K). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Neutrophil cell line (HL-60) and human umbilical vein endothelial cell (HUVEC) were gained from the American Type Culture Collection (ATCC) and grown at 37°C with 5% CO 2 .

Techniques: Flow Cytometry, Expressing, Western Blot, Immunofluorescence, Staining, Reverse Transcription, Quantitative RT-PCR, Incubation, Migration, Transwell Assay, Tube Formation Assay, Standard Deviation

Journal: Cell Reports Medicine

Article Title: Enalaprilat reverses neutrophil polarization imbalance via targeting taurine-STING axis for treatment of diabetic wounds

doi: 10.1016/j.xcrm.2026.102714

Figure Lengend Snippet: Inhibitory effects of Ena on HUVEC ferroptosis activated by AGE-elicited neutrophils (A) CCK-8 assay was performed to measure the viability of HUVECs. n = 3 independent experiments. (B) Death rate of HUVECs incubated with neutrophils pretreated by different strategies as detected using flow cytometry. n = 3 independent experiments. (C) Levels of MDA and GSH were quantified to assess the ferroptosis activity. n = 3 independent experiments. (D) Intracellular lipid peroxidation was visualized by fluorescence staining. Scale bar, 10 μm; n = 3 independent experiments. (E) Fe 2+ ion content was visualized by fluorescence staining. Scale bar, 10 μm; n = 3 independent experiments. (F) JC-1 kit was used to evaluate the mitochondrial membrane potential of HUVECs. Scale bar, 10 μm; n = 3 independent experiments. (G) DCFH-DA fluorescence probe was applied to determine ROS abundance in HUVECs. Scale bar, 200 μm; n = 3 independent experiments. (H) Mitochondrial morphology in HUVECs as analyzed by transmission electron microscopy. Scale bar, 500 nm; n = 3 independent experiments. (I) Western blot detection of PGC1α and KLF9 in HUVECs with different treatments. n = 3 independent experiments. (J) Expression of KLF9 in HUVECs incubated with neutrophils pretreated by different approaches as detected using immunofluorescence staining. Scale bar, 10 μm; n = 3 independent experiments. (K) Expression of PGC1α in HUVECs incubated with neutrophils pretreated by different approaches as detected using immunofluorescence staining. Scale bar, 10 μm; n = 3 independent experiments. (L) Schematic diagram of Ena-elicited alleviation on endothelial cell ferroptosis induced by pro-inflammatory neutrophils. Data were shown as mean ± standard deviation (SD) from biological replicates, and statistical comparisons were performed using one-way ANOVA followed by Tukey’s multiple comparisons test in (A–G and I–K). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Neutrophil cell line (HL-60) and human umbilical vein endothelial cell (HUVEC) were gained from the American Type Culture Collection (ATCC) and grown at 37°C with 5% CO 2 .

Techniques: CCK-8 Assay, Incubation, Flow Cytometry, Activity Assay, Fluorescence, Staining, Membrane, Transmission Assay, Electron Microscopy, Western Blot, Expressing, Immunofluorescence, Standard Deviation