The Effect Of P-gp In Rbmecs Under Ischemic Injury

Objective: To investigate the effect of P-gp in rBMECs under ischemic injury.Methods: Prepare the primary cultures of rBMECs、neuron cells、astrocyteand construct BBB modelsin vitro with transwell.2mM Na2S2O4was used to induce the rBMECs for4h.After treatment with Na2S2O4in rBMECs,the Papp of Rh123were measured to evaluate paracellular transport reflecting function and thePapp of NaF were measured to evaluate the quality of BBB models.The coculture of rBMECs、neuron cells、astrocytes was established to investigate the P-gp、tumour necrosis factor-α (TNF-α)、nitric oxide synthase(NOS)、protein kinase C (PKC) expression in the rBMECs under induced by Na2S2O4using Westernblot.The influence of pharmacological tools including H398,L-NMMA, BIM on the P-gp expression aswell as Rh123transendothelial transfer.Results:1.The coculture of rBMECs、neuron cells、astrocytes was established with transwell.2.Aftertreatment with2mM Na2S2O4in rBMECs for4h, the Papp of Rh123was significantly changed whichshowed that the function of P-gp can be improved.The Papp of NaF can not be significantly reduced whichshowed that interendothelial tight junction was integral.3.Western Blot analysis showed that the espressionof TNF-α、NOS can be increased as well as PKC.Incubation of H398and other pharmacological toolsdownregulated P-gp expression and functional activity in the rBMECs.Conclusions:This study suggested that TNF-α,NOS,PKC may mediate upregulation of P-gp inrBMECs under ischemic injury, which may be worthy of being referenced for the investigation of P-gp atBBB in the stroke.