Korean J Pain.  2022 Oct;35(4):433-439. 10.3344/kjp.2022.35.4.433.

Spinal orexin A attenuates opioid-induced mechanical hypersensitivity in the rat

• Youn Dh
• Jun J
• Kim TW
• Park K

• KMID: 2533983
• DOI: http://doi.org/10.3344/kjp.2022.35.4.433

Abstract

Background
Repeated administration of opioid analgesics for pain treatment can produce paradoxical hyperalgesia via peripheral and/or central mechanisms. Thus, this study investigated whether spinally (centrally) administered orexin A attenuates opioid-induced hyperalgesia (OIH).
Methods
[D-Ala² , N-Me-Phe⁴ , Gly⁵ -ol]-enkephalin (DAMGO), a selective µ-opioid receptor agonist, was used to induce mechanical hypersensitivity and was administered intradermally (4 times, 1-hour intervals) on the rat hind paw dorsum. To determine whether post- or pretreatments with spinal orexin A, dynorphin A, and antidynorphin A were effective in OIH, the drugs were injected through an intrathecal catheter whose tip was positioned dorsally at the L3 segment of the spinal cord (5 µg for all). Mechanical hypersensitivity was assessed using von Frey monofilaments.
Results
Repeated intradermal injections of DAMGO resulted in mechanical hypersensitivity in rats, lasting more than 8 days. Although the first intrathecal treatment of orexin A on the 6th day after DAMGO exposure did not show any significant effect on the mechanical threshold, the second (on the 8th day) significantly attenuated the DAMGO-induced mechanical hypersensitivity, which disappeared when the type 1 orexin receptor (OX1R) was blocked. However, intrathecal administration of dynorphin or an anti-dynorphin antibody (dynorphin antagonists) had no effect on DAMGO-induced hypersensitivity. Lastly, pretreatment with orexin A, dynorphin, or anti-dynorphin did not prevent DAMGO-induced mechanical hypersensitivity.
Conclusions
Spinal orexin A attenuates mechanical hyperalgesia induced by repetitive intradermal injections of DAMGO through OX1R. These data suggest that OIH can be potentially treated by activating the orexin A-OX1R pathway in the spinal dorsal horn.

Keyword

Analgesics; Opioid; Dynorphins; Enkephalin; Ala(2)-MePhe(4)-Gly(5)-; Hyperalgesia; Orexins; Orexin Receptors; Pain; Spinal Cord Dorsal Horn

Figure

• 

  Fig. 1 Repeated exposure to [D-Ala2, N-Me-Phe4, Gly5-ol]-enkephalin (DAMGO) induced mechanical hypersensitivity in rats. Repetitive intradermal injection (4 times, at 1-hour intervals) of DAMGO (5 µg/5 µL) into the hind paw dorsum significantly decreased mechanical threshold measured by von Frey filament test (g) on the day of and 4 and 6 days after injection. The error bars indicate SEM. **P < 0.01 vs. saline control; #P < 0.05 and ##P < 0.01 vs. baseline (i.e., before the DAMGO injection).

• 

  Fig. 2 Spinal orexin A attenuated [D-Ala2, N-Me-Phe4, Gly5-ol]-enkephalin (DAMGO)-induced mechanical hypersensitivity. Intrathecal administration of orexin A (5 µg/5 µL, i.t.) normalized the mechanical hypersensitivity that was developed on the day of and lasted until 8 days after repetitive intradermal injection (4 times, at 1-hour intervals) of DAMGO on the rat hind paw dorsum, although the first intrathecal administration of orexin A (6 days post DAMGO) had no effect. The effect of orexin A was inhibited by SB674042 (5 µg/5 µL), the OX1 receptor antagonist. The error bars indicate SEM. *P < 0.05 vs. baseline of each group; #P < 0.05 vs. the thresholds (g) of other four groups at 8 days after intradermal DAMGO injection. An experimental scheme above the histogram is shown with the number of rats used vs. the number of rats subjected with catheter insertion (n = 31/33). OX1: type 1 orexin, DMSO: dimethyl sulfoxide.

• 

  Fig. 3 Spinal dynorphin or preventing spinal endogenous dynorphin signals had no effect on the [D-Ala2, N-Me-Phe4, Gly5-ol]-enkephalin (DAMGO)-induced mechanical hypersensitivity. The mechanical hypersensitivity induced by repetitive intradermal injection of DAMGO (4 times, at 1-hour intervals) was not affected by intrathecal administration (i.t.) of dynorphin (5 µg/5 µL) or antiserum to dynorphin (5 µg/5 µL) at 6 and 8 days after the DAMGO exposure. The error bars indicate SEM. *P < 0.05 vs. baseline. An experimental scheme above the histogram is shown with the number of rats used vs. the number of rats subjected with catheter insertion (n = 18/23).

• 

  Fig. 4 Pre-dosing of orexin A or dynorphin did not prevent the induction of mechanical hypersensitivity by repeated exposure of [D-Ala2, N-Me-Phe4, Gly5-ol]-enkephalin (DAMGO). The mechanical hypersensitivity was induced by four times of repetitive intradermal injection of DAMGO (hourly). Orexin A (5 µg/5 µL), dynorphin (5 µg/5 µL), antiserum to dynorphin (5 µg/5 µL), or saline were intrathecally administered just before the DAMGO exposure. The error bars indicate SEM. **P < 0.01 vs. baseline mechanical sensitivity before DAMGO exposure. An experimental scheme above the histogram is shown with the number of rats used vs. the number of rats subjected with catheter insertion (n = 32/35). i.t.: intrathecal administration.

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