| Evidence Sentence: |
Further, 12 SCI rats were randomly divided into EA and miR subgroups (EA + miR-449a agomir injection). |
| Evidence Sentence: |
High level of miR-449a in miR subgroup was accompanied by decreased expression of Nestin, NeuN and CGRP and increased expression of cleaved caspase 3, TNF-alpha, IL-1beta and elevation of the ratio of Bax/Bcl-2 (P < 0.05), suggesting miR-449a inhibits the effects of EA on NSCs and neurons. |
| Evidence Sentence: |
3), miR-449a expression in miR group was significantly higher than that in EA group (P < 0.05). |
| Evidence Sentence: |
When compared with negative control group, miR-449a mimics significantly reduced the RLA in HEK 293T cells transfected with pMIR- CGRP-UTR (RLA = 0.47, P < 0.01), but did not affect the RLA in pMIR-CGRP-UTR-mut group (Figure 4B(Fig. |
| Evidence Sentence: |
4)), indicating that miR-449a could recognize and bind to the 3'-UTR of CGRP mRNA and regulate the CGRP expression. |
| Evidence Sentence: |
Electro-acupuncture promotes the proliferation of neural stem cells and the survival of neurons by downregulating miR-449a in rat with spinal cord injury |
| Evidence Sentence: |
MicroRNA 449a (miR-449a) expression was also compared. |
| Evidence Sentence: |
Further, 12 SCI rats were randomly divided into EA and miR subgroups (EA + miR-449a agomir injection). |
| Evidence Sentence: |
The expression of Nestin, NeuN, CGRP, cleaved caspase 3, Bax, Bcl-2, TNF-alpha, IL-1beta and miR-449a was compared. |
| Evidence Sentence: |
The direct interaction of miR-449a and CGRP mRNA was assessed by dual luciferase reporter assay. |
| Evidence Sentence: |
At day 7, compared with sham-operated group, miR-449a expression in SCI group was significantly increased (P < 0.05), and NeuN and CGRP mRNA and protein expression was markedly decreased (P < 0.05), but protein levels of Nestin, cleaved caspase 3, TNF-alpha, IL-1beta and the ratio of Bax/Bcl-2 in SCI group were significantly increased (P < 0.05). |
| Evidence Sentence: |
The EA treatment significantly reduced miR-449a level and cleaved caspase 3, TNF-alpha, IL-1beta level and the ratio of Bax/Bcl-2 (P < 0.01), but substantially increased Nestin, NeuN and CGRP expression (P < 0.05 or 0.01). |
| Evidence Sentence: |
High level of miR-449a in miR subgroup was accompanied by decreased expression of Nestin, NeuN and CGRP and increased expression of cleaved caspase 3, TNF-alpha, IL-1beta and elevation of the ratio of Bax/Bcl-2 (P < 0.05), suggesting miR-449a inhibits the effects of EA on NSCs and neurons. |
| Evidence Sentence: |
Luciferase reporter assay showed that miR-449a bound to the 3' UTR of CGRP, and thereby regulated CGRP expression. |
| Evidence Sentence: |
In conclusion, EA promotes proliferation of neural stem cells and the survival of neurons by downregulation of miR-449a expression. |
| Evidence Sentence: |
Effects of EA on Nestin, NeuN, CGRP and miR-449a expression |
| Evidence Sentence: |
The miR-449a expression in SCI group was significantly increased compared with that in sham-operated group (P < 0.05), whereas miR-449a expression was significantly reduced after EA treatment (P < 0.01, Figure 1A(Fig. |
| Evidence Sentence: |
Effect of miR-449a agomir on EA-stimulated Nestin, NeuN and CGRP expression and EA-reduced level of cleaved caspase 3, TNF-alpha and IL-1beta and the ratio of Bax to Bcl-2 |
| Evidence Sentence: |
Further, the regulation of miR-449a on EA-stimulated Nestin, NeuN and CGRP expression was assessed by comparing rats receiving EA treatment and EA+miR-449a agomir treatment. |
| Evidence Sentence: |
3), miR-449a expression in miR group was significantly higher than that in EA group (P < 0.05). |
| Evidence Sentence: |
These results suggest that high level of miR-449a could suppress the effect of EA on the expression of Nestin, NeuN, CGRP, cleaved caspase 3, Bax, Bcl-2, TNF-alpha and IL-1beta in spinal cord of rats. |
| Evidence Sentence: |
miR-449a regulates CGRP expression by directly binding to its 3'UTR |
| Evidence Sentence: |
The regulation of miR-449a on the CGRP expression was further determined by Luciferase reporter assay. |
