| Evidence Sentence: |
The PSD-95 protein level was up-regulated in the small interfering (si)RNA-IL16 group compared to the NC-IL16 groups with or without oxygen/glucose deprivation/reperfusion (OGD/R) conditions (P<0.05). |
| Evidence Sentence: |
Western blot analysis revealed PSD-95 protein levels were increased in the si-IL16 group compared to the NC-IL16 group in or not in OGD/R conditions (P<0.05) (Figure 6), but the overall levels of both were reduced in OGD/R conditions (P<0.01). |
| Evidence Sentence: |
NC-IL16 + NC-mimic + NC-inhibitor + OGD group), but in the absence of IL-16, PSD-95 was downregulated in cells treated with miR-81 mimic (i.e., NC-IL16 + mimic + NC-inhibitor + OGD vs. |
| Evidence Sentence: |
Si-IL16 + mimic + NC-inhibitor + OGD; P<0.05). |
| Evidence Sentence: |
IL-16 is a potential target of miR-81 |
| Evidence Sentence: |
In this study, we investigated the effect of EA at the Baihui (GV 20) and Shenting (GV 24) acupoints on spatial learning and memory ability, and the expression level of microRNA-81 (miR-81), interleukin-16 (IL-16), and postsynaptic density protein-95 (PSD-95) in the frontal cortex of VaD rats. |
| Evidence Sentence: |
Immunochemistry (IHC), quantitative reverse transcription polymerase chain reaction (qRT-PCR), and western blot analysis were performed to detect the expression level of miR-81, IL-16, and PSD-95. |
| Evidence Sentence: |
Finally, luciferase assay was used to determine the effect of miR-81 on IL-16 expression in PC12 cells. |
| Evidence Sentence: |
In the PC12 cell validation experiment, IL-16 expression level was reduced under the condition of the miR-81 mimic treatment, while increased in the miR-81 inhibitor group. |
| Evidence Sentence: |
Binding sites in the miR-81 sequence (5'-GUCAGUG-3') and the IL-16 gene sequence (5'-CAGUCAC-3') were predicted via bioinformatics analysis (Figure 5A). |
| Evidence Sentence: |
To evaluate the direct effects of miR-81 on IL-16 gene expression, we constructed a dual luciferase reporter carrying the miR-81 target site in pLUC-IL-16 (Figure 5B). |
| Evidence Sentence: |
The data indicated that the miR-81 mimic decreased the luciferase activity of IL-16. |
| Evidence Sentence: |
Contrastingly, the miR-81 mimic did not decrease activity of luciferase while the miR-81 seed sequence of the IL-16 3'UTR was mutated (Figure 5C). |
| Evidence Sentence: |
To further assess the relationship between miR-81 and IL-16, PC12 cells were divided into 4 groups: miR-81 mimic, miR-81 mimic-NC, miR-81 inhibitor, and miR-81 inhibitor-NC. |
| Evidence Sentence: |
After transfecting PC12 cells with miR-81 mimic, miR-81 mimic-NC, miR-81 inhibitor, and miR-81 inhibitor-NC, the relative expression of IL-16 mRNA was measured using RT-qPCR. |
| Evidence Sentence: |
Overall, IL-16 expression was decreased in the mimic group compared to the mimic-NC group and increased in the inhibitor group compared to the inhibitor-NC group (Figure 5E). |
| Evidence Sentence: |
These results indicate that miR-81 directly regulates IL-16. |
| Evidence Sentence: |
To determine whether IL-16 mediates the effect of miR-81 on regulation of PSD-95, IL-16 expression was silenced in PC12 cells subjected to oxygen glucose deprivation/reperfusion (OGD/R) injury and followed by miR-81 mimic/inhibitor treatment. |
| Evidence Sentence: |
NC-IL16 + NC-mimic + NC-inhibitor + OGD group), but in the absence of IL-16, PSD-95 was downregulated in cells treated with miR-81 mimic (i.e., NC-IL16 + mimic + NC-inhibitor + OGD vs. |
| Evidence Sentence: |
These results indicate that IL-16 is required for the effects of miR-81 for the regulation of PSD-95 in OGD/R injury. |
| Evidence Sentence: |
Interleukin-16 mediates the effect of miR-81 on regulation of PSD-95 |
| Evidence Sentence: |
Electroacupuncture of the Baihui and Shenting acupoints for vascular dementia in rats through the miR-81/IL-16/PSD-95 pathway |
| Evidence Sentence: |
In VaD rats, EA may improve spatial learning and memory through miR-81/IL-16/PSD-95 pathway. |
