Puerarin Nanostructured Lipid Carriers For Preparation And Aβ _25-35 Inducedy PC12 Cell Research

Objective To prepare Puerarin-loaded nanostructured lipid carrier （Pue-NLC）andinvestigate their physicochemical properties. To investigate the pharmacokinetics of Pue-NLCand the neuroprotective effects on the apoptosis of in rat pheochromocytoma PC12reduced byAβ_25-35.Method Pue-NLC prepared by the method of emulsion-ultrasonic dispersion, and theformula and technology were optimized by the single factor exploration and orthogonal design.The particle morphology of dispersion was observed by transmission electron microscopy.Zetapotential and particle size analyzer was determined surface potentials,size and distribution.Theentrapment efficiency was determined by centrifugation ultrafiltration method.The in vitro drugreleasing behavior was studied by dialysis. The formulation of lyophilized Pue-NLC wasoptimized based on its appearance, color and dispersing properties. Pue solution and Pue-NLCwere injected of tail vein of rats,the plasma concentration was detected by the HPLCmethod.The cell model of Alzheimer’s disease （AD） was set up by Aβ_25-35-induced toxicityPC12cells.The effect of Pue solution and Pue-NLC on cell proliferation was detected by MTTassay,and the effect of that on cell apoptosis was detected by flow cytometry.Results Selected optimization prescription through the orthogonal experiment,underthe optimal conditions,morphology of Pue-NLC is round and smooth, average particle size（89±7） nm. Entrapment efficiency was （91.33±1.2）%.The Zeta potential was （-22±0.4） mV, andit has a good release characteristics in vitro.To determine plasma concentration of the rats in thedifferent time,Obtained a series of pharmacokinetic parameters and Pue-NLC has good slow-release ability.After i.v. injection into tail venin of rats AUC,MRT and T1/2of Pue-NLC in liverwas1.75,1.80and3.09times higher than Pue solution. MTT assay revealed that Aβ_25-35atconcentration of20.00μmol·L^-1was employed induced PC12cells effective concentration.Pretreatment of10.00μmol·L^-1Pue solution and Pue-NLC for24h significantly decreasedcell death induced by20.00μmol·L^-1Aβ_25-35.Flow cytometry was performed to analyze cellapoptosis results revealed the number of apoptosis cell significantly increased in Aβ_25-35treatment group compared with blank group,and statistical significance.Compared with Aβ_25-35treatment group,pretreatment of PC12cells with Pue solution and Pue-NLC significantlydecreased cell apoptosis that Pue-NLC were much better than Pue solution. Conclusion Pue-NLC prepared by using the method of emulsification ultrasonicdispersion meets the requirements of the pharmaceutics. Experimental results in vivo indicatethat the Pue-NLC slows the release of the drug.The results of in vitro cell test have shown thatsurvival rate and protective action of Pue-NLC on Aβ_25-35were higher than the concentration ofPue solution.