| Evidence Sentence: |
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. |
| 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: |
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: |
Effects of EA on Nestin, NeuN, CGRP and miR-449a expression |
| Evidence Sentence: |
While the expression of Nestin protein in SCI group was to some extent higher than that in sham-operated group. |
| Evidence Sentence: |
We found no significant difference in Nestin expression between the two groups. |
| Evidence Sentence: |
When compared with SCI group, the Nestin, NeuN and CGRP mRNA and protein expression in SCI+EA group was significantly increased (P < 0.05 or 0.01). |
| Evidence Sentence: |
The immunohistochemical results also showed that Nestin protein level was raised to some degree, but NeuN and CGRP protein levels were markedly declined in SCI group (P < 0.01 or P < 0.05), which were all raised in SCI+EA group (P < 0.05 or 0.01) (Figure 1E-H(Fig. |
| Evidence Sentence: |
1)), suggesting that SCI might elevate the Nestin level but reduce the NeuN and CGRP level in spinal cord of rats, and EA treatment after SCI would elevate the level of Nestin, NeuN and CGRP in ventral horn of spinal cord of rats. |
| 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: |
Moreover, the results of real time PCR, Western blot and immunohistochemical staining were also shown that the expression of Nestin, NeuN and CGRP mRNA and protein in miR group was significantly reduced compared with EA group (P < 0.05 or 0.01) (Figure 3B-H(Fig. |
| 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. |
