The effect of eriocitrin on lipopolysaccharide-induced inflammation of endothelial cells in sepsis by regulating SphK1/S1P pathway

Zhang Kaiya; Xu Yaoyao; Yang Qin; Li Xia

doi:10.3969/j.issn.1002-1949.2025.02.011

Abstract：Objective To study the repair effect of eriocitrin on lipopolysaccharide(LPS)-induced inflammation of endothelial cells in sepsis,and to explore the mechanism of eriocitrin in endothelial cells in sepsis based on the sphingosine kinase 1/sphingosine-1-phosphate(SphK1/S1P)pathway.Methods Human umbilical vein endothelial cells(HUVEC)were divided into blank control group,model group,low-dose,medium-dose and high-dose eriocitrin groups and positive group.The model group,low-dose,medium-dose and high-dose eriocitrin groups and positive group were added with final concentration of 100 ng/mL LPS for 24 h,and the sepsis model was prepared.The low-dose,medium-dose and high-dose groups were added with the final concentration of 100,200 and 400 µg/mL eriocitrin,the positive group was added with the final concentration of 10 μmol/L of new psoralen isoflavones,the blank control group and the model group were added with the culture medium.Levels of nitric oxide(NO),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),granulocyte-macrophage colony-stimulating factor(GM-CSF),monocyte chemoattractant protein-1(MCP-1),interferon-γ(IFN-γ),interleukin-8(IL-8)in HUVEC supernatant and cellular myeloperoxidase(MPO),superoxide disumutase(SOD),malondialdehyde(MDA),C-reactive protein(CRP)and lactate dehydrogenase(LDH)were determined by enzyme-linked immunosorbent assay(ELISA)kit,transmembrane resistance(TER)was determined by transmembrane resistance meter,and HUVEC permeability was determined by fluorescence labeling method.The apoptotic rate of HUVEC was determined by apoptosis kit.The mRNA and protein levels of SphK1 and S1P of HUVEC in different groups were determined by real-time quantitative polymerase chain reaction(PCR)and Western blot,respectively.Results Compared with blank control group,HUVEC supernatant levels of NO,IL-6,TNF-α,GM-CSF,MCP-1,IFN-γ and IL-8,MPO,MDA,CRP and LDH,HUVEC permeability and apoptosis rate,the mRNA and protein levels of SphK1 and S1P were increased in model group,SOD and TER were decreased,the difference was statistically significant(P＜0.05).Compared with model group,HUVEC supernatant levels of NO,IL-6,TNF-α,GM-CSF,MCP-1,IFN-γ and IL-8,MPO,MDA,CRP and LDH,HUVEC permeability and apoptosis rate,and the mRNA and protein levels of SphK1 and S1P in low-dose,medium-dose and high dose eriocitrin groups and positive group were decreased,while SOD and TER were increased,the difference was statistically significant(P＜0.05).Conclusions Eriocitrin can significantly inhibit the inflammation of endothelial cells in sepsis induced by LPS,improve the oxidative damage of endothelial cells in sepsis induced by LPS,repair the permeability of endothelial cells in sepsis,and then inhibit the apoptosis of endothelial cells in sepsis.The mechanism may be related to the regulation of SphK1/S1P pathway.