In:
CNS Neuroscience & Therapeutics, Wiley, Vol. 20, No. 1 ( 2014-01), p. 76-85
Abstract:
Gene therapy targeting the SNCA gene yields promising results in the treatment of Parkinson's disease ( PD ). The most challenging issue of the RNA i gene therapy strategy is maintaining efficient delivery without inducing significant toxicity and other adverse effects. This study aimed to characterize polyethylene glycol‐polyethyleneimine as a vector for alpha‐synuclein si RNA delivery to PC12 cells for Parkinson's disease. Methods The characteristics of PEG‐PEI /si SNCA were analyzed via gel retardation assay and assessments of particle size and zeta potential. MTT cytotoxicity assay and flow cytometry were used to detect cytotoxicity and transfection efficiency in PC12 cells. Confocal laser scanning microscopy was employed to examine the intracellular distribution of PEG‐PEI / FITC ‐si SNCA after cellular uptake. RT‐PCR and western blotting were used to measure SNCA expression. The MTT cytotoxicity assay was used to study the effect of PEG‐PEI /si SNCA on cell viability. The protective effect of PEG‐PEI /si SNCA on MPP +‐induced apoptosis in PC12 cells was examined via flow cytometry and Hoechst staining. Results PEG‐PEI /si SNCA complexes were well‐developed; they exhibited appropriate particle sizes and zeta potentials at a mass ratio of 5:1. In vitro , PEG‐PEI /si SNCA was associated with low cytotoxicity and high transfection efficiency. Complexes were capable of successfully delivering si SNCA into PC12 cells and releasing it from the endosome. Furthermore, PEG‐PEI /si SNCA could effectively suppress SNCA m RNA expression and protected cells from death via apoptosis induced by MPP + . Conclusions Our results demonstrate that PEG‐PEI performs well as a vector for alpha‐synuclein si RNA delivery into PC12 cells. Additionally, PEG‐PEI /si SNCA complexes were suggested to be able to protect cells from death via apoptosis induced by MPP + . These findings suggest that PEG‐PEI /si SNCA nanoparticles exhibit remarkable potential as a gene delivery system for P arkinson's disease.
Type of Medium:
Online Resource
ISSN:
1755-5930
,
1755-5949
DOI:
10.1111/cns.2013.20.issue-1
Language:
English
Publisher:
Wiley
Publication Date:
2014
detail.hit.zdb_id:
2423467-9
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