The Study Of Synergistic Effect Combination Drug Yi-Qi-Jie-Du On Acute Ischemic Stroke Based On The Vitro Model Of Neurovascular Unit

Ischemic stroke is a cerebral vascular and high incidence disease that serious threat to the health and life of human,with characteristics of high incidence,high mortality,high morbidity and high recurrence rate.At present,the single target drugs and screening model can not meet the therapeutic purposes of complex disease,this study adopts the neurovascular unit integration target,to research the anti-brain damage synergy effect of Yi-Qi-Jie-Du formula(YJ).Objective The establishment of cell level drug-evaluation platform neurovascular unit(NVU).The anti-ischemic stroke effect research on combination drug YJ based on neurovascular unit.The anti-ischemic stroke synergy effect study of YJ.Methods Using extracted rat primary cortical cells and rat microvascular endothelial cells(rBMEC)to establish the NVU model in vitro,and identified the three kinds cells and the tight junction protein Claudin-5 via the immunofluorescence.To investigate the reasonable inducing concentration of damaging agents on NVU,and the relationship of effective concentration dependence.Conducting signal component,two-two and YJ party administration based on different time and different concentration to evaluation the YJ synergistic pharmacodynamic effect.Results1.Establish and standardize the construct method of NVU model,through the verification on the structure and function of it to prove that the model establishment is successful.2.Determined the damage agent and its administration concentration is 1 mM/L hydrogen peroxide and 5 mM/L L-glutamate joint inducing the NVU.The effective concentration 2 μM/L,10 μM/L,50 μM/L all could explore the TEER,FICT-BSA permeability,Sodium Fluorescein permeability,Alkaline Phosphatase activity,Myeloperoxidase activity,Monoamine oxidase activity in a dose-dependent relationship.3.The results of the signal component,two two,YJ party administration in different concentration on different time points shows the effect the two two administration is better compare with signal administration,the YJ party administration compared with the single drug and two two administration,express the best effect.Conclusions1.In this paper,the established NVU model in vitro could simulate the blood brain barrier(BBB)function in vivo,and has the characteristics of multi-targets and system integration.The NVU model associated the neuro protection with the vascular protection dynamically,had obvious superiority compared with the single target and single cell model,especially suitable for complicated system of traditional Chinese Medicine research.The study provided effective means for the encephalopathy prevention and the bioactivity screening and evaluation of raditional Chinese medicine compound in vitro.2.For the first time endue the substantive content to the NVU conceptual model,and established the system of quantitative index of dynamic monitoring.The model has been applied to the synergistic effect research of anti-encephalopathy drug YJ.Through the evaluation results multi-index evaluation system shows the ginsenosides,berberine,gardenia party administration express the strongest effect and has obvious synergistic effect,and the geniposide may play an important role in the synergistic effect of the YJ formula.