| Objective:To prepare CHP self-assembled nanoparticles.Donepezil CHP nanoparticles(DZP-CHP)were prepared with donepezil as a model drug and characterized by a series of physical and chemical properties.To investigate the brain targeting of DZP-CHP nanoparticles and to explore its protective effect on Aβ25-35-induced PC12 and SH-SY5Y cell injury,and to provide a new theoretical basis for the anti-AD study of DZP-CHP nanoparticles.Methods:The DZP-CHP nanoparticles were prepared by water dialysis.The morphology of nanoparticles was observed by transmission electron microscopy and scanning electron microscopy.The particle size and Zeta potential were determined by dynamic light scattering.The entrapment efficiency and drug loading were measured by ultraviolet spectrophotometry.Dialysis assay in vitro release characteristics.Live Fluorescence Imaging System was used to observe the distribution of nanoparticles in mice.The cell model of AD was established by inducing PC12 and SH-SY5Y cells with Aβ25-35.The effects of DZP solution and DZP-CHP Nanometer solution on the cell viability were detected by MTT assay.The content of lactate dehydrogenase(LDH)in the culture supernatant was measured by spectrophotometry,Rhodamine 123 was used to detect the change of mitochondrial potential and AO-EB staining was used to detect the morphological changes of apoptosis.Results:DZP-CHP nanoparticles were prepared by water dialysis.Transmission electron microscopy and scanning electron microscopy showed that the blank CHP nanoparticles and DZP-CHP nanoparticles were spherical with uniform morphology and uniform size.The average particle size is about 260nm,the dispersion coefficient is less than 0.2,the surface of particle is negatively charged,and has good drug release characteristics in vitro.The loading and entrapment efficiency of donepezil with CHP were better,and the best ratio was 1:5.In vivo fluorescence imaging system results show that the surface modified Tween80 DZP-CHP nanoparticles with brain-targeting.Incubation of PC12 and SH-SY5Y cells with different concentrations of Aβ25-35 for 24 h resulted in a significant decrease of cell proliferation,increased LDH of supernatant and Aβ25-35 induced injury of PC12 and SH-SY5Y cells at 20μmol·L-1 was effective concentration.Compared with Aβ25-35 in control group,pretreatment of cells with DZP solution at 5,10μmol·L-1 concentration and DZP-CHP solution for 2 hours significantly improved the cell proliferation activity induced by Aβ25-35.The decrease of LDH release and the increase of mitochondrial potential,and the effect of DZP-CHP nanometer solution on Aβ25-35-induced apoptosis was better than that of DZP solution group.Conclusion:The DZP-CHP nanoparticle with surface-modified polysorbate 80 is brain-targeted and the protective effect of DZP-CHP nanoparticle on PC 12 and SH-SY5Y cells induced by Aβ25-35 is higher than that of DZP solution. |
