Objective To elucidate the underlying mechanism of Panax notoginseng saponin(PNS)on gastric epithelial cell injury and barrier dysfunction induced by dual antiplatelet(DA).Methods Human gastric mucosal epithelial cell...Objective To elucidate the underlying mechanism of Panax notoginseng saponin(PNS)on gastric epithelial cell injury and barrier dysfunction induced by dual antiplatelet(DA).Methods Human gastric mucosal epithelial cell(GES-1)was cultured and divided into 4 groups:a control,a DA,a PNS+DA and a LY294002+PNS+DA group.GES-1 apoptosis was detected by flow cytometry,cell permeability were detected using Transwell,level of prostaglandins E2(PGE2),6-keto-prostaglandin F1α(6-keto-PGF1α)and vascular endothelial growth factor(VEGF)in supernatant were measured by enzyme linked immunosorbent assay(ELISA),expression of phosphatidylinositide 3-kinase(PI3K),phosphorylated-PI3K(p-PI3K),Akt,phosphorylated-Akt(p-Akt),cyclooxygenase-1(COX-1),cyclooxygenase-2(COX-2),glycogen synthase kinase-3β(GSK-3β)and Ras homolog gene family member A(RhoA)were measured by Western-blot.Results DA induced apoptosis and hyper-permeability in GES-1,reduced supernatant level of PGE2,6-keto-PGF1αand VEGF(P<0.05).Addition of PNS reduced the apoptosis of GES-1 caused by DA,restored the concentration of PGE2,6-keto-PGF1αand VEGF(P<0.05).In addition,PNS attenuated the alteration of COX-1 and COX-2 expression induced by DA,up-regulated p-PI3K/p-Akt,down-regulated RhoA and GSK-3β.LY294002 mitigated the effects of PNS on cell apoptosis,cell permeability,VEGF concentration,and expression of RhoA and GSK-3βsignificantly.Conclusions PNS attenuates the suppression on COX/PG pathway from DA,alleviates DA-induced GES-1 apoptosis and barrier dysfunction through PI3K/Akt/VEGF-GSK-3β-RhoA network pathway.展开更多
基金Supported by the National Natural Science Foundation of China(No.81273933 and 81102722)the Eleven Five-Year Plan of National Science and Technology Support Project(No.2006BAI04A01-2)the Jilin Province Major Science and Technology Achievement Transforming Project(No.11ZDZH005)。
文摘Objective To elucidate the underlying mechanism of Panax notoginseng saponin(PNS)on gastric epithelial cell injury and barrier dysfunction induced by dual antiplatelet(DA).Methods Human gastric mucosal epithelial cell(GES-1)was cultured and divided into 4 groups:a control,a DA,a PNS+DA and a LY294002+PNS+DA group.GES-1 apoptosis was detected by flow cytometry,cell permeability were detected using Transwell,level of prostaglandins E2(PGE2),6-keto-prostaglandin F1α(6-keto-PGF1α)and vascular endothelial growth factor(VEGF)in supernatant were measured by enzyme linked immunosorbent assay(ELISA),expression of phosphatidylinositide 3-kinase(PI3K),phosphorylated-PI3K(p-PI3K),Akt,phosphorylated-Akt(p-Akt),cyclooxygenase-1(COX-1),cyclooxygenase-2(COX-2),glycogen synthase kinase-3β(GSK-3β)and Ras homolog gene family member A(RhoA)were measured by Western-blot.Results DA induced apoptosis and hyper-permeability in GES-1,reduced supernatant level of PGE2,6-keto-PGF1αand VEGF(P<0.05).Addition of PNS reduced the apoptosis of GES-1 caused by DA,restored the concentration of PGE2,6-keto-PGF1αand VEGF(P<0.05).In addition,PNS attenuated the alteration of COX-1 and COX-2 expression induced by DA,up-regulated p-PI3K/p-Akt,down-regulated RhoA and GSK-3β.LY294002 mitigated the effects of PNS on cell apoptosis,cell permeability,VEGF concentration,and expression of RhoA and GSK-3βsignificantly.Conclusions PNS attenuates the suppression on COX/PG pathway from DA,alleviates DA-induced GES-1 apoptosis and barrier dysfunction through PI3K/Akt/VEGF-GSK-3β-RhoA network pathway.
