Protective Effects And Its Mechanism Of MPEG-Cyclovirobuxine And Cyclovirobuxine

The cerebrovascular disease （CVD）, such as stroke already has been one of the diseases that badlyhazardous health. The characteristic of stroke are high incidence rate、high death rate and highrecrudescence rate. The most badly result is that it can induce sequela, which can bring heavily burdento his family and himself. In recent years, the research to cerebrovascular disease has benn obtainin-depth develop. It is make sure that cyclovirobuxine has obviously protective effect to bothcerebrovascular disease and heart disease. This study on the basis of prophase research, throughexperiment to discuss the protective effect of mPEG-Cyclovirobuxine to reperfusion injury.Part oneProtective Effect of mPEG-Cyclovirobuxine and Cyclovirobuxine on MACO ratsObjective To observe the effect of mPEG-CVB and CVB on MCAO rats and then investigate themechanism. Methods The model of MCAO rats was produced by transitory occlusion of the left middlecerebral artery（MCA） with nylon line. After 4 hours, the line was pulled out and artery was reirrigatedfor 20 hours. The behavior scores of each rat, cerebral infaction, water content and the index of brainwere investigated, in order to study the protective effect of mPEG-CVB.Results The results indicatedthat mPEG-CVB and CVB could attenuate behavior scores, reduce cerebral infaction and water content,and the influence of mPEG-CVB is obvious than CVB in the same dose. Conclusion mPEG-CVB couldreform the hydrocephalus in MACO rats, which related to reducing the putrescence of nerve cell.,this isone of important mechanisms for their effectiveness in cerebral ischemic disease.Part two Effects of mPEG-Cyclovirobuxine and Cyclovirobuxine on MCAO rats’microcirculatory blood flowObjective To study the mechanism of mPEG-CVB and CVB in treating cerebral diseases byobserveing its influence on MCAO rats’ microcirculatory blood flow. Methods The observation on theeffect of drugs on the cerebral microcirculatory blood flow was performed with laser microcirculatorydynamic analyzer. Results The cerebral microcirculatory blood flow decreased after MCAO,Intravenous injection of mPEG-CVB and CVB could increase the cerebral microcirculatory blood flow,and the influence of mPEG-CVB is obvious than CVB in the same dose. Conclusion mPEG-CVB canincrease cerebral blood flow in rats with MCAO, which is one of important mechanisms for theireffectiveness in cerebral ischemic disease.Part threeDetermination of mPEG-cyclovirobuxine through the Blood-Brain-Barrier by HPLCObjective To study the relation of mPEG-CVB and Blood-Brain-Barrier by establishing a new HPLCmethod for the determination of mPEG- CVB in the rat brain. Methods Pretreatment brainsample which is contain durg, Kromasic C18 column with temperature 35℃was used. Themobile phase consisted of absolute methanol-water（30:70）, and was set at a flow rate of 0.8ml·min^-1.Evaporative light scattering detector was used. Results mPEG-CVB existence in the sample,but lack of stability, have some differentia. Conclusion The ability of mPEG-CVB through the BBB innot tranquilization and should study the way to help more mPEG-CVB penetrate across the BBBPart fourEffects of mPEG-Cyclovirobuxine and Cyciovirobuxine on Intracellular Free CalciumConcentration in Neonatal Rats Cardiac CellsObjective To measure free Ca²⁺ concentration of cultured cardiac cells of neonatal rats,and observe theeffect of mPEG-CVB and CVB on [Ca²⁺]i. Methods Using Fura-2/AM and Fluo-3/AM as Ca²⁺ sensitiveprobe, to observe the change of intracellular Ca²⁺ by double-wavelengh spometerand confocal laserscanning micrectrophotoscopy （CLSM） Results The value of Ca²⁺ of cardiac muscle was decrease alsoin mPEG-CVB and CV-B group, in a connentraction-dependent manner.there was significant difference between model and mPEG-CVB group （P＜0.05, P＜0.01）, and the influence of mPEG-CVB is obviousthan CVB in the same dose. Conclusion Double-wavelengh spectrophotometer and CLSM system canaccutately reflect cytosolic concentration of free Ca²⁺ labeld by Fura-2/AM and Fluo-3/AM,mPEG-CVB can decrease [Ca²⁺]i in cultured cardiac cells, which provides a direct evidence for alleviateanoxia/reoxygenation injury mechanism.