Electro-acupuncture promotes the proliferation of neural stem cells and the survival of neurons by downregulating miR-449a in rat with spinal cord injury

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Abstract

The aim of this study is to investigate the mechanism of electro-acupuncture (EA) on the recovery of injured spinal cord. Rats were randomly divided into normal control, sham-operated, SCI, SCI+EA group and T10 segment spinal cord injury (SCI) rat model was established by the modified Allen's method. After 7 days, the mRNA and protein expression of Nestin, neuron specific nuclear protein (NeuN) and calcitonin gene related peptide (CGRP) were detected by real time RT-PCR, Western blot and immunohistochemistry respectively. The protein expression of cleaved caspase 3, Bax, Bcl-2, TNF-α and IL-1β were also detected by Western blot. MicroRNA 449a (miR-449a) expression was also compared. Further, 12 SCI rats were randomly divided into EA and miR subgroups (EA + miR-449a agomir injection). The expression of Nestin, NeuN, CGRP, cleaved caspase 3, Bax, Bcl-2, TNF-α, IL-1β and miR-449a was compared. The direct interaction of miR-449a and CGRP mRNA was assessed by dual luciferase reporter assay. At day 7, compared with sham-operated group, miR-449a expression in SCI group was significantly increased (P P P P P P < 0.05), suggesting miR-449a inhibits the effects of EA on NSCs and neurons. Luciferase reporter assay showed that miR-449a bound to the 3' UTR of CGRP, and thereby regulated CGRP expression. In conclusion, EA promotes proliferation of neural stem cells and the survival of neurons by downregulation of miR-449a expression.

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