【Animal modeling】-The improvement effect of Xueshuantong capsule on tight junction injury of HUVEC cells induced by oxygen-glucose deprivation/reoxygenation

  Objective: To investigate the protective effects of Panax notoginseng saponins (PNS) and its monomers Rg1-Rb1 on the damage of vascular endothelial tight junctions induced by oxygen-glucose deprivation (OGD) in a model of HUVEC cells induced by oxygen-glucose deprivation (OGD).

  Methods: HUVEC cells were seeded in Transwell cell chambers, and after cell fusion, experimental groups were established: normal control group, model group, PNS 10, 20, and 40 mg/L group, and Rb1 13.52 mg/L, Rg1 12.44 mg/L group. , drug intervention in hypoxia, oxygen and glucose deprivation for 6 h and reoxygenation for 24 h, the changes of HUVEC endothelial cell resistance and endothelial permeability were detected; laser confocal microscopy and immunofluorescence were used to detect HUVEC cells ZO-1?claudin -5 Changes in protein expression.

  Results: Compared with the normal control group, the resistance of tight junctions between endothelial cells in HUVEC cells was significantly decreased and the cell permeability was significantly increased after 6 h of hypoxia and 24 h of reoxygenation (P < 0.05). PNS 20 and 40 mg/L groups could significantly inhibit the decrease of tight junction resistance and the increase of HUVEC cell permeability in the model group (P < 0.05). Ginsenoside Rb1 could significantly increase the resistance of endothelial tight junctions and decrease the permeability of HUVEC cells after 6 h reoxygenation of OGD for 24 h (P < 0.05). The results of immunofluorescence showed that the ZO-1 and claudin-5 proteins of HUVEC cells in the normal control group were mainly distributed around the cell membrane and arranged continuously, showing the typical arrangement of paving stones of endothelial cells. After 6 h of hypoxia and 24 h of reoxygenation, the tight junction protein in the cell edge of the model group was significantly reduced, the ring structure was broken or even disappeared, and the tight junction protein in the nucleus increased. At PNS 20, 40 mg/L and Rb1 13.52 mg/L, local recovery of tight junctions between cells could be observed, showing a paving stone-like structure.

  Conclusion: PNS has a protective effect on ischemia-induced tight junction injury between vascular endothelial cells.