The Construction Of ZiyuglycosideⅠ-AuNPs And Its Effects On Autophagy In Hematopoietic Stem Cells

Objective:To construct the Ziyuglycoside I（ZgⅠ）-AuNPs and explore its effects on autophagy in hematopoietic stem cells.Methods:（1）Prescription study of ZgⅠ-AuNPs:the optimal formulation of ZgⅠ-AuNPs was studied,including the preparation method of AuNPs,the drug loading method,the ratio of HAu Cl₄ and modified materials.Finally,the apperance,particle size and UV specific absorbance were evaluated.（2）Quality study of ZgⅠ-AuNPs:ZgⅠ-AuNPs,ZgⅠ,AuNPs and their physical mixtures were analyzed by XRD and DSC,to explore the existing form of ZgⅠ in AuNPs.The appearance,morphology,spatial structure and particle size of ZgⅠ-AuNPs were observed by TEM and SEM.The composition and structure of ZgⅠ-AuNPs were confirmed by FTIR.Furthermore,the drug loading of ZgⅠ in AuNPs and the in vitro release behavior were studied.（3）The effects of ZgⅠ-AuNPs on autophagy in hematopoietic stem cells:the CTX was used to induce the injury of hematopoietic stem cell in vitro,after the intervention of ZgⅠ-AuNPs,the autophagy state of hematopoietic stem cells was detected by TEM,and the autophagy symbolic protein LC3B was detected by immuneluorescene.Results:（1）AuNPs were prepared by HAu Cl₄-sodium citrate reduction method.The optimized ZgⅠ-AuNPs formula,(HAu Cl₄（1%w/w）,sodium citrate（38.8m M）,SH-PEG-NH₂（0.25 m M）,ZgⅠ amount(100μg·m L^-1)),was the most convenient one with drug content of 15.63μg/mg.（2）The TEM and SEM examinations showed that ZgⅠ-AuNPs had an irregular cubic crystal structure with an average particle size of340 nm.The XRD,DSC and FTIR detections proved that ZgⅠ was successfully loaded into the composite system.Besides,the in vivo evaluation showed that ZgⅠ-AuNPs had slower drug release rate.（3）ZgⅠ-AuNPs had low toxic to hematopoietic stem cells and significantly inhibited the autophagy reduce induced by CTX.Conclusion:In this experiment,a simple,stable and repeatable preparation method of ZgⅠ-AuNPs was established,which can obtain ZgⅠ-AuNPs with good appearance,high drug loading and drug release properties.What’s the most important,after modification with AuNPs,the promotion of ZgⅠ on autophagy of the hematopoietic stem cells was significantly enhanced,suggesting that AuNPs-modified nanoparticles loading Ziyuglycoside I has potential application in myelosuppression therapy.