Fluorescent Silica Nanoparticles Charged With 10-23DRz Inhibit The Expression Of HBV

OBJECTIVE:Chronic Hepatitis B(CHB) is a serious disease of harming the human health.At present,there are no effective drugs to cure it.Recently,scientists are trying to carry out gene therapy for its treatment.CHB can cause body immune injury through viral gene expression.The introduction of nanoparticle vector promotes the development of gene therapy.This research aims at preparing the fluorescent silica nanoparticles(FSNPs),probing into the feasibility of FSNPs as a gene transfer vector and the easy detection of the fluorescence of FSNPs as a biology marker,studying FSNPs-10-23 DRz inhibition of the expression of HBsAg and HBeAg,exploring the possibility of FSNPs-10-23 DRz complex treating CHB at cellular and molecular levels.Partâ… The preparation and biological characteristics of FSNPsMETHODS:The FSNPs were prepared by microemulsion.NaCl was used to do surface modification on them.After a process of drying, their size was detected by a particle analyzer,and their Surface Zeta Potential was measured by an electrical Potential analysator.The FSNPs were dissolved in the physiological saline and their physiological saline solution was administered of the SD rats through the tail vein.Its acute toxicity to the SD rats was detected by up-and-down procedure and its distribution in the main organs of the SD rats was observed with fluorescence microscope.RESULTS:The FSNPs were successfully prepared in the W/O microemulsion.The nanoparticles were spherical and uniform in size, about 32 nm in diameter,+53.9mv in surface Zeta potential and highly monodispersed.More than 2000mg/kg LD₅₀ of the FSNPs was estimated by up and down procedure.Under the observation of fluorescence microscope,the FSNPs emitted red fluorescence distributed in each of the main organs.This indicated that the FSNPs could successfully pass through the blood-brain and blood-prostate barriers.PARTâ…¡The FSNPs deliver 10-23DRz to inhibit the expression of HBVMETHODS:The special 10-23DRzs were designed by the sequence of S and C gene of HBV.The special 10-23DRzs was binded to the modified nanoparticles.The binding effect was tested by 15% denaturing polyacrylamide gel electrophoresis.Adding the FSNPs-DNA complex into fetal bovine serum,the protective effect of nanoparticles was detected by 15%denaturing polyacrylamide gel electrophoresis.The HepG2.2.15 cell was incubated with FSNPs-DNA complex,its inhibitory effect to HBsAg and HBeAg was analyzed by ELISA assay.RESULTS:The FSNPs could combine DNA effectively,protect against nuclease degradation in the serum.The NANO-DNA complex can inhibit the expression of S and C gene of HBV.The inhibition rate was 53.9mv and was higher than the liposomes control group.Conclusion:The FSNPs were successfully prepared,and it could combine special 10-23DRz and this complex could enter HepG2.2.15 cell to inhibit the expression of S and C gene of HBV.