The Effect Of T19, A Acridine Compounds, On The P-gp Function And Expression Under H₂O₂ Induced In RBMECs

Objective:To investigate the effect of acridine compound T19 on the P-glycoprotein?P-gp?function and expression under hydrogen peroxide?H₂O₂?induced in rat brain microvascular endothelial cells?rBMECs?.Methods:Cultured the rBMECs and astrocytes by primary culture,and MTT assay was used to measure the effect of T19 on the cell viability.The coculture of rBMECs and astrocytes was prepared with transwell technology,the Papp of NaF was measured to evaluate the effect of T19 on the status of permeability of the rBMECs monolayer.The Papp of Rh123 was measured to evaluate the influence of T19on the function of P-gp.After treatment with the different concentration of H₂O₂ in rBMECs,the Papp of NaF and Rh123 was measured to screen the condition of oxidative stress model.After treatment with T19and H₂O₂,the transendothelial transfer of Rh123 and NaF was measured to reflect the effect of T19 on the P-gp function under H₂O₂ induced.ROS and the distribution of Nrf2 was detected by flow cytometry and confocal microscopy,respectively.The protein expression of P-gp?P-ERK?P-I B were detected by Western Blot.ERK inhibitor PD98059 and NF-B inhibitor PDTC were used to evaluate the influence of inhibitors on the P-gp function and determine the relationship between ERK and NF-B under H₂O₂induced.Results:The effect of T19 on proliferation and tight junction had not obviously changed.T19 reduced basal P-gp function and expression in concentration-dependent manner.In order to maintain consistent with the experimental results in vivo,treatment with 150?M H₂O₂ for 4 h to establish oxidative stress model.T19 protects rBMECs from oxidative stress by enhancing Nrf2 translocation.T19 decrease H₂O₂-induced the change of P-gp function and expression in rBMECs,through inhibited the activation of ERK and NF-B.Conclusions:T19 reduced P-gp expression under H₂O₂ induced through inhibited the activation of ERK and NF-B.T19 protects rBMECs from oxidative stress by enhancing Nrf2 translocation.