Salvianolic Acid B Regulates Ang ?-induced Vascular Endothelial Cell Oxidative Stress Upon AKT,MAPK Signaling Pathway

ObjectiveTo establish the oxidative stress model of Human umbilical vein endothelial cells(HUVEC)induced by Angiotensinogen II(Ang II),observe the effect of Salvianolic acid B(SalB)intervention,and analyze the process of inhibition of the oxidative stress response.The pharmacological mechanism of SALB was also discussed in this present study.Methods1.MTT method was used to screen the non-toxic intervention concentration of SalB to HUVEC cells,then Ang II was used to induce oxidative stress in HUVEC cells.The reactive oxygen species(ROS),nitric oxide(NO)and the intracellular calcium concentration were detected with corresponding fluorescent probes,and malondialdehyde(MDA)content was detected with TAB method.2.Western blot was used to detect the expression levels of p-ERK1/2,ERK1/2,p-JNK?JNK,p-p38,p38,PI3 K,p-AKT,AKT,iNOS,p-NRF2,NRF2,HO-1 and NOX2 proteins in HUVEC.Results1.MTT assay showed that 25 ?M,50 ?M,100 ?M SalB intervene HUVEC had no significant difference in cell viability,indicating that the concentration was no toxic to HUVEC.Compared with the Control group,the intracellular ROS level and MDA content of HUVEC increased significantly after 1 ?M Ang II induction,while the NO content decreased significantly.Compared with Ang II group,after SalB pretreatment,intracellular ROS and MDA content significantly decreased,NO content and intracellular calcium concentration release increased.2.Western Blot showed that 1 ?M Ang II could upregulate the expression of p-ERK,P-P38 and P-JNK,and downregulate the expression of NOX2,PI3 K,P-Akt,P-NRF2 and HO-1.pretreatment of SalB significantly upregulated the phosphorylation levels of ERK1/2,P38,JNK,PI3 K,AKT,NRF2 and HO-1 protein expression in HUVEC cell,and inhibited the expression of NOX2 protein.Conclusions1.SalB can play an antioxidant role by inhibiting the increase of Ang II-induced ROS and MDA content and increasing the level of NO in HUVEC cells.2.SalB can further activate the release of calcium ion concentration in HUVEC cells stimulated by Ang II.3.SalB can further enhance the activation of PI3K/AKT and MPAK(ERK1/2,P38,JNK)signaling pathways induced,inhibit HUVEC oxidative stress response induced by Ang II by activating NRF2 / HO-1 signaling pathway,reduce endothelial cell damage,and thus protect endothelial cells.