Protective effects and mechanisms of breviscapine on endothelial cells / 中国药科大学学报

ABSTRACT

This study was to investigate the protective effects and mechanisms of breviscapine on endothelial cells.ox-LDL was used to induce oxidative damage on human umbilical vein endothelial cells(HUVEC)for 20 h with or without 4 h pretreatment of various concentrations of breviscapine(10;20;40 μmol/L);then observe the protective effect and mechanism of breviscapine on endothelial cells insulted by ox-LDL.MTT method was used to detect cell viability;flow cytometry was used to detect cell apoptosis and reactive oxygen species;and Western blot and RT-PCR was used to detect cell signaling pathways.The results showed that breviscapine recovered HUVEC cell viability in a dose-dependent manner which inhibited by ox-LDL;and it also protected cell from apoptosis induced by ox-LDL.To explore the mechanisms of breviscapine;reactive oxygen species (ROS)was determined after pretreatment of various concentrations of breviscapine or different durations(2;4;6 h)of brevis-capine.Results showed that breviscapine decreased ROS production in a dose-and time-dependent manner.Fur-thermore;cell signaling pathway analysis showed that breviscapine increased the expression of BCL-2;decreased the expression of BAX and the release of Cytochrome C and cleavage of caspase-3.Breviscapine decreased Keap1 and activated the nuclear import of Nrf2;and subsequently increased the mRNA and protein expression of down-stream antioxidant enzyme as NAD(P)Hquinone oxidoreductase 1(NQO1)and glutathione transferase-S-Mu 1(GSTM1);and increasing the activity of NQO1.Besides;breviscapine decreased IKK and IKB;and inhibited nuclear translocation of NF-κB;while increasing the expression of eNOS.This study demonstrated that breviscap-ine has a protective role on ox-LDL-induced endothelial cell injury;which may be related to its antioxidant effects and inhibition of NF-κB activation.