Effects Of Panax Notoginseng Saponins On Expression Of PLA2G4A And Its Downstream Products In Cerebral Microvascular Endothelial Cells Induced By Ischemic Injur

Panax Notoginseng Saponins(PNS)is an active ingredient extracted from traditional Chinese medicine Panax notoginseng.Recently,it has been found that Panax Notoginseng Saponins can effectively inhibit the development of cerebral ischemia,and improve the cerebral microvasc?Lar coag?Lation and blood flow.There are a lot of studies on the mechanism of Panax Notoginseng Saponins can improve the cerebral blood flow after ischemia.However,the upstream molec?Lar mechanism of is still to be further explored.The research by a gene chip detection,found that after ischemic injury of brain microvasc?Lar endothelial cells(Brain Microvasc?Lar Endothelial Cells,BMECs),cytosolic phospholipase A2IVA(phospholipaseA2groupIVA,PLA2G4A)mRNA overexpression significantly,and Panax Notoginseng Saponins has a significant down reg?Late effect on PLA2G4A mRNA.PLA2G4A is a reg?Latory enzyme widely exists in cell metabolism,its specific decomposition of cell membrane phospholipids,and generate a variety of downstream metabolites,these products can reg?Late the blood coa?Lation and vasc?Lar contractile function.Therefore,we spec?Late that the pathological process of PLA2G4A mRNA expression of cerebral ischemia and cerebral vasc?Lar hemodynamic relevance,and inhibition of PLA2G4A mRNA expression may be a mechanism of total Panax Notoginseng Saponins reg?Late cerebral ischemia cerebral blood flow after cerebral ischemia.Therefore,this experiment on the basis of former research,design experimental to observate Panax Notoginseng Saponins on ischemic injury of endothelial cells to reg?Late PLA2G4A mRNA and its downstream products,in order to explore the molec?Lar mechanism of Panax Notoginseng Saponins in improving of cerebral blood flow.Objective:To investigate the effects of Panax Notoginseng Saponins on PLA2G4A gmRNA and protein and its downstream products in ischemic cerebral microvasc?Lar endothelial cells,to explore the mechanism of the effect of Panax Notoginseng Saponins on ischemic cerebral blood flow,to provide the basis for the improvement of blood flow after cerebral ischemia by Panax Notoginseng Saponins,and provide new pharmacological targets and experimental basis for the prevention and treatment of ischemic cerebrovasc?Lar diseases.Methods:1.Isolation and c?Lture of rat primary MMECs and establishment of invitro oxygen-glucose-dEPrivation(OGD)model.2.To observe the expression of PLA2G4A mRNA and protein in brain microvasc?Lar endothelial cells after ischemic injury:RT-PCR and Western Blotting were used to detect the expression of PLA2G4A mRNA and protein in brain microvasc?Lar endothelial cells of PNS group,model group and blank group after ischemic injury3.Screening of PLA2G4A inhibitor concentration,Pyrrophenone(Cayman)can specifically inhibit the activity of PLA2G4A and inhibit the production of arachidonic acid(ArachidonicAcid,AA),The third generation rat brain micro vasc?Lar cells were used as experimental objects.Set several concentration gradient on OGD-induced BMECs.ELISA assay was used to detect the AA production of endothelial cells after administration.Calc?Lated the half inhibitory concentration(IC50)of Pyrrophenone on AA in the OGD-induced BMECs4.Observe the effect of Panax Notoginseng Saponins on the products of arachidonic acid,which was induced by ischemic injury in brain microvasc?Lar endothelial cells,BMECs was divided into normal group,OGD model group,PNS group and inhibitor group.After treatment,the AA content in each group was detected by Elisa method5.Observe the effects of Panax Notoginseng Saponins on 20-HETE,TXA2 andPGI2 in the brain microvasc?Lar endothelial cells,BMECs was divided into normal group,OGD model group,PNS group and inhibitor group,After treatment,the 20-HETE,TXA2 and PGI2 content in each group was detected by Elisa method.Res?Lts:1.We observed that PLA2G4A expression in brain microvasc?Lar endothelial cells.2.Oxygen glucose dEPrivation can increase the expression of PLA2G4A mRNA and protein in brain microvasc?Lar endothelial cells;compared with the normal group,the PLA2G4A mRNA and protein levels in the OGD group were significantly increased3.PNS significantly inhibited the expression of PLA2G4A mRNA and protein in brain microvasc?Lar endothelial cells induced by OGD,from the res?Lts of RT-PCR and Western Blot we know,compared with the model group,the expression of PLA2G4A mRNA and protein in PNS group was significantly reduced after OGD,the difference was statistically significant.4.PNS inhibited OGD induced AA production.By Elisa method,we can see that compared with the model group,PNS down reg?Lated the expression of AA induced by OGD,the difference was statistically significant.5.PNS can reduce the expression of TXA2,PGI2,20-HETE in brain microvasc?Lar endothelial cells after ischemic injury.ELISA res?Lts showed that compared with the normal group,the expression of TXA2,PGI2,20-HETE in the model group was significantly increased.the TXA2,PGI2 and 20-HETE of the PNS group decreased significantly compared with the model group,the difference was statistically significant.Conclusion:1.In this study,PLA2G4A expression in rat brain microvasc?Lar endothelial cells was observed,The levels of PLA2G4AmRNA and protein were significantly increased after OGD modeling.PNS can inhibit the increase of PLA2G4A mRNA and protein level caused by the OGD modeling.2.PNS can effectively down reg?Late the over expression of AA,TXA2,PGI2 and 20-HETE after modeling.This may be related to the inhibition of PLA2G4A mRNA and protein production by PNS.This may be one of the mechanisms by which PNS plays an important role in improving cerebral blood flow in ischemic brain injury.