Lentivirus-Mediated Overexpression of miR-29a Promotes Axonal Regeneration and Functional Recovery in Experimental Spinal Cord Injury via PI3K/Akt/mTOR Pathway

Neurochemical Research
Hua YinJin-Bo Liu

Abstract

MicroRNAs as a novel class of endogenous small non-coding RNAs, modulate negative gene expression at the post-transcriptional level. Our previous work has demonstrated that miR-29a reduces PTEN expression by directly targeting the 3'-UTRs (untranslated regions) of its mRNA, thus promoting neurite outgrowth. To further confirm the role of miR-29a in the recovery of SCI and its potential mechanisms, a recombinant lentiviral vector was used to promote miR-29a expression in the injured spinal cord. As compared with the LV-eGFP group and normal saline group, a significantly increased level of miR-29a expression and a markedly decreased level of PTEN expression were observed in the LV-miR-29a group. Overexpression of miR-29a increased the phosphorylation of two proteins (Akt and S6) of PI3K-AKT-mTOR signaling pathway and the expression of axonal regeneration associated key marker protein (neurofiament-200). Moreover, quantitative imaging analysis was performed to confirm that LV-miR-29a group expressed axonal regeneration at 4.0 ± 0.2-fold as much as the other two groups. Besides, miR-29a overexpression promoted hindlimb motor functional recovery. Collectively, these results suggested that miR-29a may be an important regulator for ax...Continue Reading

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Methods Mentioned

BETA
PCR
ELISA
Assay
protein assay
Electrophoresis
transfection

Software Mentioned

Image J
Total Lab
SDS
GraphPad Prism

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