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Phosphorylated p38 MAPK (phospho-p38 MAPK) in the lumbar spinal cord was quantified using western blotting. |
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Phospho-p38 MAPK nuclear translocation in spinal dorsal horn was examined using immunohistochemical staining and confocal microscopy. |
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Inhibition of SP signaling pathway or activity of p38 MAPK significantly improved the EA analgesia In EA poor-responders with SCD, which provides a promising way to treat the chronic pain in patients with SCD. |
| Evidence Sentence: |
Quantitative Western blot analysis of whole lumbar spinal cords showed that combined treatment with EA and netupitant in both moderate- and non-responders (Non/Moderate-responders + EA + Netupitant) significantly reduced the expression of phospho-p38 MAPK relative to total-p38 MAPK (Figure 3A, B). |
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Nuclear co-localization of phospho-p38 MAPK in the dorsal horn of the spinal cord was highly insightful because nuclear translocation of p38 MAPK is critical in the transcriptional regulation of inflammatory cytokines. |
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We observed a significantly high co-localization of phospho-p38 MAPK immunoreactivity in the nulei of non-responders (red and white dots indicated by yellow arrows in the cyan nuclei) compared to high- responders (Figure 3C, D). |
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Similarly, moderate-responders also showed significantly higher nuclear presence of phospho-p38 MAPK compared to high-responders. |
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Either netupitant or EA alone did not have any effect on nuclear phospho-p38 MAPK in non- or moderate-responders. |
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Interestingly, poor responders exhibited high levels of substance P (SP), a mediator of chronic pain, as well as active p38 MAPK in spinal cords. |
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The present study aimed to investigate the roles of inhibition of SP and SP-activated p38 MAPK in chronic pain in sickle mice that are poorly responsive to EA intervention (moderate/non-responders). |
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or p38 MAPK inhibitor (SB203580, 10 mg/kg/day, i.p.) |
| Evidence Sentence: |
Phosphorylated p38 MAPK (phospho-p38 MAPK) in the lumbar spinal cord was quantified using western blotting. |
| Evidence Sentence: |
Results: In EA poor-responders, combined treatment with EA and netupitant significantly enhanced the analgesic effects of EA in poor-responders on mechanical, heat, cold, and deep tissue pain, and decreased phosphorylation of p38 MAPK in lumbar spinal cords and its nuclear translocation in the spinal dorsal horn. |
| Evidence Sentence: |
Quantitative Western blot analysis of whole lumbar spinal cords showed that combined treatment with EA and netupitant in both moderate- and non-responders (Non/Moderate-responders + EA + Netupitant) significantly reduced the expression of phospho-p38 MAPK relative to total-p38 MAPK (Figure 3A, B). |
| Evidence Sentence: |
Nuclear co-localization of phospho-p38 MAPK in the dorsal horn of the spinal cord was highly insightful because nuclear translocation of p38 MAPK is critical in the transcriptional regulation of inflammatory cytokines. |
| Evidence Sentence: |
However, combined treatment with EA and netupitant significantly inhibited p38 MAPK phosphorylation as well as its nuclear translocation. |
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High-responders did not show significant p38 MAPK phosphorylation upon Western blotting (Figure 3A and B) or upon immunofluorescent co-localization in the cytoplasm and/or nuclear translocation (Figure 3C, D). |
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Together these data suggest that the non-responsiveness to EA in non- and moderate-responders may be due to increased spinal p38 MAPK phosphorylation due to higher NK1R activity perhaps due to increased SP levels in these sickle mice. |
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Conclusions: These results suggest a pivotal role of SP in maintaining the chronic pain in SCD via spinal phospho-p38 MAPK signaling, which may hinder the effect of EA in poor responders. |