Effect Of Gas6 On Endothelial Barrier Induced By Pg-Lps

To explore the function of Growth-arrest-specific protein 6 (Gas6) on vascular endothelial barrier integrity induced by Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) in human umbilical vein endothelial cells (HUVECs). HUVECs were cultured until they reached confluence and subsequently stimulated with Pg-LPS. Then, the endothelial barrier permeability was assessed by FITC-Dextran trans-endothelial permeability assay. After up-regulation and down-regulation of the expression of Gas6, HUVECs were stimulated with Pg-LPS. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was taken to examine the expression of intercellular junctions, including zonula occludens-1 (ZO-1), occludin, and adherens junction vascular endothelial-cadherin (VE-cadherin). The protein expression levels of intercellular junctions were detected by Western blot. Immunofluorescence was taken to detect the distribution of junction proteins in the cell–cell junction areas. The concentration of FITC-Dextran in the lower chamber of the Transwell changed significantly after the endothelial cells were stimulated with Pg-LPS for 4 h, and the endothelial barrier permeability further increased with time. Gas6 overexpression inhibited the decreased expression of intercellular junctions induced by Pg-LPS. While the reduction in the expression of intercellular junctions induced by Pg-LPS was promoted in down-regulation group. Furthermore, Gas6 overexpression or knockdown influenced the distribution of the junction proteins under Pg-LPS stimulation. Changing the expression of Gas6 can affect the expression and distribution of intercellular junctions after Pg-LPS stimulating, suggesting that Gas6 may play a role on endothelial barrier integrity.