GSK484,a PAD4 inhibitor,improves endothelial dysfunction in mice with sepsis-induced lung injury by inhibiting H3Cit expression
Objective To investigate the inhibitory effect of GSK484,a PAD4 inhibitor,on H3Cit expression following sepsis and its effects for improving sepsis-induced endothelial dysfunction.Methods Eighteen C57BL/6 mice were randomized into sham-operated group,sepsis model group and GSK484 treatment group(n=6),and in the latter two groups,models of sepsis were established by cecal ligation and puncture(CLP).The mice in GSK484 treatment group were given an intraperitoneal injection of GSK484(4 mg/kg)on the second day following the surgery.Twenty-four hours after the injection,the mice were euthanized for measurement of serum levels of VEGF,ESM-1,IL-6 and IL-1β using ELISA.Lung tissue pathology was observed with HE staining,and pulmonary expressions of F-actin,VE-cadherin,ZO-1 and H3Cit proteins were detected using immunofluorescence staining and Western blotting.In primary cultured of mouse lung microvascular endothelial cells,the effect of stimulation with LPS(10 μg/mL)for 24 h on tube formation,proliferation,apoptosis and expressions of VEGF,ESM-1,IL-6 and IL-1β were assessed using CCK-8 assay,flow cytometry and ELISA.Results Compared to the sham-operated mice,the septic mice exhibited significant lung tissue pathologies characterized by vascular congestion,alveolar rupture,edema,and neutrophil infiltration.Serum levels of IL-6,IL-1β,VEGF,and ESM-1 were elevated,pulmonary expressions of F-actin,VE-cadherin,and ZO-1 were decreased,and H3Cit expression was increased significantly in the septic mice.GSK484 treatment effectively mitigated these changes in the septic mice.The LPS-stimulated endothelial cells showed increased productions of IL-6,IL-1β,VEGF and ESM-1,which were significantly reduced after treatment with 2.5 pmol/L GSK484.Conclusion GSK484 treatment effectively suppresses H3Cit expression in septic mice to ameliorate sepsis-induced endothelial dysfunction.
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维普
