Exploration on the effect and mechanism of ginsenoside Rb1 in ameliorating hypoxia/reoxygenation-induced endothelial cell leakage based on mitochondrial ATP synthesis function
Objective:To investigate whether ginsenoside Rb1 protects against hypoxia/reoxygenation(H/R)injury-induced HUVEC barrier dysfunction and its related mechanisms.Methods:HUVEC cells were used to establish a H/R injury model by continuous hypoxia 2 h/reoxygenation 2 h.A control group,a Rb1 background group,a model group,an Rb1 intervention group(1×10-3,1×10-4,1×10-5 mol/L),and an Rb1+oligomycin group were set up.The permeability of the endothelial cell monolayer was assessed using FITC-Dextran tracer method,and the ATP expression level in cells was evaluated using fluorescent probe labeling and ATP assay kit.The expression levels of ZO-1,Occludin,and Claudin-5 were detected by immunofluorescence and Western Blot.The morphology of mitochondria in cells was observed using mito-Tracker Red CMXRos fluorescent probe method,and the activities of mitochondrial respiratory chain complexes Ⅰ and Ⅴ were determined by ELISA.Results:Compared with the model group,1 × 10-4 mol/L Rb1 intervention group significantly ameliorated the increased permeability of HUVEC cells after reoxygenation(P＜0.05)and increased cellular ZO-1,Occludin and Claudin-5 protein expression(P＜0.05),1 × 10-4 mol/L Rbl intervention group significantly increased intracellular ATP levels(P＜0.05),improved the activity of mitochondrial respiratory chain complexes Ⅰ and V(P＜0.05),and markedly reduced cellular mitochondrial division status.The permeability of the endothelium was elevated in the Rb1+oligomycin group compared with the 1 × 10-4 mol/L Rb1 intervention group(P＜0.05),and the expression of ZO-1,Occludin,and Claudin-5 proteins were all reduced(P＜0.05).Conclusion:Ginsenoside Rb1 exerts a protective effect on the endothelial barrier by improving the structure and function of mitochondria in endothelial cells after H/R,increasing ATP synthesis,and enhancing the expression of endothelial cell junction proteins.
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