Objective To prepare fluorescent silica nanoparticles(FSNPs) carrying 10-23 deoxyribozymes(10-23DRzs) and to evaluate their inhibitory effect on human hepatitis B virus(HBV).Methods The FSNPs were prepared by microemu...Objective To prepare fluorescent silica nanoparticles(FSNPs) carrying 10-23 deoxyribozymes(10-23DRzs) and to evaluate their inhibitory effect on human hepatitis B virus(HBV).Methods The FSNPs were prepared by microemulsion method and further modified by NaCl.Their diameter and the Zeta potential were tested.The HBV-specific 10-23DRzs were designed according to the sequences of S and C genes of HBV.The 10-23DRzs were connected to FSNPs,which constituted the FSNPs-DNA.The connecting efficiency and the protective effect of nanoparticles on 10-23DRzs were tested by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE).The HepG2.2.15 cells were transfected by FSNPs-DNA,of which the inhibitory effects on HBsAg and HBeAg were analyzed by enzyme-linked immunosorbent assay(ELISA).Results The nanoparticles were spherical and uniform in size,with a diameter of 220 nm and the surface Zeta potential of +15.2 mV.The combination of DNA and FSNPs effectively protected DNA from nuclease degradation.Transfection of FSNPs-DNA significantly inhibited the expression of HBV S and C genes compared to the liposome control group.Conclusion The FSNPs have been successfully prepared and efficiently connected to HBV-specific 10-23DRzs,which significantly inhibit the expression of HBV S and C genes in cell culture.展开更多
基金Supported by National Hi-tech Project of China(No.2007AA02-1803 and 2007AA021901)
文摘Objective To prepare fluorescent silica nanoparticles(FSNPs) carrying 10-23 deoxyribozymes(10-23DRzs) and to evaluate their inhibitory effect on human hepatitis B virus(HBV).Methods The FSNPs were prepared by microemulsion method and further modified by NaCl.Their diameter and the Zeta potential were tested.The HBV-specific 10-23DRzs were designed according to the sequences of S and C genes of HBV.The 10-23DRzs were connected to FSNPs,which constituted the FSNPs-DNA.The connecting efficiency and the protective effect of nanoparticles on 10-23DRzs were tested by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE).The HepG2.2.15 cells were transfected by FSNPs-DNA,of which the inhibitory effects on HBsAg and HBeAg were analyzed by enzyme-linked immunosorbent assay(ELISA).Results The nanoparticles were spherical and uniform in size,with a diameter of 220 nm and the surface Zeta potential of +15.2 mV.The combination of DNA and FSNPs effectively protected DNA from nuclease degradation.Transfection of FSNPs-DNA significantly inhibited the expression of HBV S and C genes compared to the liposome control group.Conclusion The FSNPs have been successfully prepared and efficiently connected to HBV-specific 10-23DRzs,which significantly inhibit the expression of HBV S and C genes in cell culture.