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Objective To investigate the interventional effect ofsalvianolic acid B(SalB)on activated platelet-induced inflammatory response in human brain microvascular endothelial cells (HBMECs).Methods Activated platelets and HBMECs were co-cultured to establish inflammatory response models.The mRNA expression levels of inflammatory mediators,including intercellular adhesion molecule-1(ICAM-1),IL-1β,IL-6,IL-8 and monocyte chemoattractant protein-1(MCP-1)in HBMECs,were detected by quantitative RT-PCR.Cells of the pretreatment group were pretreated with 10 μg/ml SalB for 24 h,and the changes of mRNA expression levels of these inflammatory mediators were analyzed.Results The mRNA expression levels ofICAM-1,IL-1β,IL-6,IL-8 and MCP-1 in activated platelet-induced inflammatory response model of SalB pretreatnent group were significantly reduced as compared with those without pretreatment(P＜0.05).Conclusion SalB can repress the activated platelet-induced inflammatory response in HBMECs,which might be one of the mechanisms of anti-cerebral ischemia effect.
RESUMEN
Objective To preliminarily study the role of activated platelets in inflammatory response in ischemic stroke by using the platelet and vascular endothelial cell co-culture model.Methods Platelets were isolated from healthy donator's peripheral blood and activated in vitro by 0.8 μmol/L ADP in ischemic stroke; resting platelets were also chosen.The mean platelet component (MPC)concentration was measured by hematology analyzer.Resting platelets and activated platelets were co-cultured with vascular endothelial cells for 12 h,respectively; the other 2 control groups (ADP only,blank) were employed in the experiment.RT-PCR was used to detect the expression level of mRNA of proinflammatory cytokines in the endothelial cells,including IL-1β,IL-6,IL-8 and MCP-1.Results Compared with the resting platelets[(266.80±22.14) g/L],ADP-induced activated platelets[(231.90±14.63) g/L]showed significantly lower concentration of MPC (P<0.05).Activated platelets in the co-culture model could induce the mRNA expression of IL-1β,IL-6,IL-8 and MCP-1 in the endothelial cells,while resting platelets could not.Conclusion Activated platelets can trigger an interaction between platelets and endothelial cells in vitro,and induce the mRNA expression of proinflammatory cytokines in the endothelial cells,which may be one of the mechanisms of inflammatory response in iscbemic stroke.