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: Expression vectors encoding ZNF460, GGT1, KLF9 and PGC1α were constructed by GeneChem (Shanghai, China) via cloning the open reading frames of the indicated genes into a pcDNA3.1 plasmid vector.
Techniques: CCK-8 Assay, Incubation, Flow Cytometry, Activity Assay, Fluorescence, Staining, Membrane, Transmission Assay, Electron Microscopy, Western Blot, Expressing, Immunofluorescence, Standard Deviation