The Reasearch Of Ginsenoside Compound K On Neurons Injuried By Acute Ischemic Stroke In Vitro

Objective: Acute Ischemic Stroke(AIS)refers to the local brain tissue because of the blood circulation disorder,which is ischemia and hypoxia result in necrosis,with corresponding symptoms and signs,has become the first disability and the second cause of death in the world.The incidence of AIS accounted for about 80% of stroke.The key treatment of AIS is recover the infarct zone blood supply actively,and save the brain tissue of ischemic penumbra area.At present,people take the thrombolytic therapy to treat AIS,but because of constraining the short time window,and with large artery embolism recanalization rate is low,so did not achieve ideal trentment effect.Ginsenoside Compound K is an intestinal bacterial metabolite of protopanaxadiol type saponins,plays practical role in human body.Ginsenoside Compound K could enhance the release of neurotransmitters,improve brain blood supply,scaveng free radical,antioxidant and so on.The unique pharmacological activity of Ginsenoside Compound K has already caused the extensive concern,and its clinical range of applications is increasingly.This experiment selects the cortical neurons of neonatal Sprague Dawley(SD)rats as the research object,through hypoxic regulation system and neuronal sugar-free culture medium to construct Oxygen and glucose deprivation/reoxygenation and recovery of glucose model.At the same time,select the extract of Ginkgo biloba(GBE)as the positive control drug,observe the intervention of Ginsenoside Compound K on neurons injuried by OGD/R by two kinds of different administration,which ispreconditioning and acute administration,to further test the effect of Ginsenoside Compound K on AIS,and to provide scientific experimental basis for clinical medicine.Methods: 1.Extract cortical neurons of neonatal SD rat,through Microtubule-associated protein 2(MAP2)by immunofluorescence staining in neurons and DAPI nuclear staining,observe morphological characteristic and identify purity of the cells.2.Through hypoxic regulation system and neuronal sugar-free culture medium to construct OGD/R model,adoptting pretreatment and acute administration two routes of administration,through MTT test,LDH test and TUNEL test,observe the effect of Ginsenoside Compound K on neurons injuried by OGD/R.Setting different drug dose,finding the best drug dose of CK on neurons.Results: 1.The results of cell survival rate by MTT colorimetry: Compared with Control group,the cell survival rate of group OGD and OGD/R decreased obviously,and with the duration of reoxygenation,cell survival rate was lower,the cell viability of group Ginsenoside Compound K enhanced obviously(P<0.05).By the pretreatment and acute administration,the best drug dose of Ginsenoside Compound K on neurons injuried by OGD/R are 10?g/ml and 1?g/ml,respectively.2.The result of LDH leakage rate: Compared with Control group,the LDH leakage rate of group OGD and OGD/R increased significantly,and with the duration of reoxygenation,the LDH leakage rate increased futher,but except for group OGD2h/R30 min,the LDH leakage rate of group Ginsenoside Compound K reduced obviously(P<0.05).3.The results of apoptosis neurons by TUNEL: The number of apoptosis neurons of group OGD and OGD/R is much more than Control group,and with the duration of reoxygenation,the rate of apoptosis neurons was more,the rate of apoptosis neurons of group Ginsenoside Compound K reduced significantly(P<0.05).Conclusion: 1.After OGD/R treatment,cell viability decreased,and with the duration of reoxygenation,cell survival rate was lower.2.By the pretreatment and acute administration,Ginsenoside Compound K can significantly exert protective effect on neurons injuried by OGD/R,and the the best drug dose are 10?g/ml and 1?g/ml,respectively.