The Protection Of Resolvin D1on LPS-induced Barrier Disruption Of Endothelial Cells

Objective: The purpose of this research is to study the protection of Resolvin D1onLPS-induced barrier disruption of endothelial cells.Methods: Lipopolysaccharide (400ng/ml) was stimulated human unbilical vein endothelialcell. Pretreatment with Resolvin D1(100ng/ml) to endothelial cell for30minutes beforetreating with LPS, the other groups were pretreated with PD98059or PDTC for30minutes,then treated with Resolvin D1and LPS for6h. Endothelial cells permeability wasmeasured by FITC–Dextran in the lower chamber. Immunofluorescence was used to surveythe redistribution of zo-1and occludin and the formation of stress fibers of actincytoskeleton. Western blot was examine the expressions of zo-1, occludin, p-ERK1/2andIκBα.Results：(1) Compared with the control group, endothelial permeability wassignificantly increased in LPS-stimulated group (P<0.01). Compared with the LPS group,endothelial permeability was decreased in RvD1group (P<0.01). Compared with the LPSgroup, the permeability of endothelial cells was downregulated by RvD1+LPS treatment(P<0.01).(2) In the control group, occludin and zo-1were distributed along the cells, F-actin wasorganized in peripheral dense bands, which made the shape of the cells stability. Comparedwith the control group, actin cytoskeleton was disrupted, stress fiber formation wasobserved; zo-1and occludin fragmentation was detected; gaps were observed byLPS-simulated group. Compared with the LPS group, the distribution of zo-1, occludin andF-actin were closed to the normal by RvD1+LPS treatment.(3) Compared with the control group, the expression of occludin and zo-1were significantly downregulate by LPS stimulated group (P<0.01), moreover, the expression.of IκBαwas decreased (P<0.01). Compared with the LPS group, the expreassion ofzo-1, occludin (P<0.01) and IκBα(P<0.05) were closed to the normal by RvD1+LPStreatment. There was no difference of P-ERK1/2between each group. Compared with theLPS+RvD1group, the expreassion of occludin was increased by LPS+RvD1+PDTCtreatment.Conclusion：RvD1protects LPS-induced barrier function through upregulated expressionof TJ proteins in HUVECs, which is relative to NF-κB pathway but not ERK1/2.