Silenced NgR gene expression by RNA interference to promote rats facial nerve regeneration in vitro.
- Author:
Yong SHI
;
Liang ZHOU
;
Jie TIAN
;
Yang WANG
- Publication Type:Journal Article
- MeSH:
Animals;
Cell Differentiation;
Cells, Cultured;
Facial Nerve;
surgery;
GPI-Linked Proteins;
genetics;
metabolism;
Myelin Proteins;
genetics;
metabolism;
Neural Stem Cells;
cytology;
metabolism;
Nogo Receptor 1;
RNA Interference;
Rats;
Rats, Sprague-Dawley;
Receptors, Cell Surface;
genetics;
metabolism
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2014;28(10):728-730
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To suppress NgR gene expression in neural stem cells and observe differentiation of neural stem cells in vitro after interfered which provide nutritional support for the facial nerve repair in vivo.
METHOD:PCR amplification, restriction endonuclease digestion, T4DNA ligase connections were used to connected NgR with rector pGCsi, and constructed recombinant vector (NgR shRNA). Lipofectamine 2000 were used to transfect the NSC. The expression of NgR was examined by Western Blot. The proportion of neural stem cells transformed into neurons after transfection was tested by Immunocytochemistry. Neural stem cells were planted in PLGA tubes after transfected, and were scanned by electron microscopy.
RESULT:NgR shRNA plasmid was constructed and infected neural stem cells successfully. Western Blot showed that the expression of NgR decreased in neural stem cells after interference. Immunocytochemistry showed that the rate of the neural stem cells transformed into neurons after interfered was significantly higher (P < 0.01).
CONCLUSION:Neural stem cells were transformed into neurons after NgR shRNA plasmid infected neural stem cells, which promoted axonal regeneration more effectively and provided a efficient and stable gene platform for facial nerve repair.
