Anesth Pain Med.  2018 Jan;13(1):82-92. 10.17085/apm.2018.13.1.82.

Differential expression of spinal γ-aminobutyric acid and opioid receptors modulates the analgesic effects of intrathecal curcumin on postoperative/inflammatory pain in rats

• Ju J
• Shin JY
• Yoon JJ
• Yin M
• Yoon MH

• KMID: 2436061
• DOI: http://doi.org/10.17085/apm.2018.13.1.82

Abstract

BACKGROUND
Curcumin is traditionally used as an herbal medicine. We explored the efficacy of intrathecal curcumin in relieving both postoperative and inflammatory pain and elucidated the mechanisms of action of curcumin interacting with γ-aminobutyric acid (GABA) and opioid receptors at the spinal level.
METHODS
Experimental pain was induced in male Sprague-Dawley rats via paw incision or injection of intraplantar carrageenan. After examination of the effects of intrathecal curcumin on the pain, GABA and opioid receptor antagonists were intrathecally administered to explore the involvement of GABA or opioid receptors on the effect of curcumin. Additionally, the expression levels of the GABA and opioid receptors were assessed.
RESULTS
Intrathecal curcumin reduced the withdrawal threshold of both incisional surgery- and carrageenan injection-induced nociception. Intrathecal GABA and opioid receptor antagonists reversed the curcumin-mediated antinociception. Incisional surgery decreased the levels of the GABA receptors mRNA, but little changed the levels of the opioid receptors mRNA. Carrageenan injection increased the levels of the opioid receptors mRNA, but not the GABA receptors mRNA levels. Intrathecal curcumin increased or decreased the levels of GABA receptors mRNA and opioid receptors mRNA in the spinal cords of incised or carrageenan-injected rats, respectively.
CONCLUSIONS
Intrathecal curcumin was effective to postoperative and inflammatory pain and such antinociception of curcumin was antagonized by both GABA and opioid receptor antagonists. Also, intrathecal curcumin altered the levels of GABA and opioid receptors. Thus, spinal GABA and opioid receptors may, respectively, be directly or indirectly involved when curcumin alleviates postoperative and inflammatory pain.

Keyword

Curcumin; γ-aminobutyric acid and opioid receptor; Postoperative and inflammatory pain; Spinal cord

MeSH Terms

Animals
Carrageenan
Curcumin*
gamma-Aminobutyric Acid
Herbal Medicine
Humans
Male
Narcotic Antagonists
Nociception
Rats*
Rats, Sprague-Dawley
Receptors, GABA
Receptors, Opioid*
RNA, Messenger
Spinal Cord
Carrageenan
Curcumin
Narcotic Antagonists
RNA, Messenger
Receptors, GABA
Receptors, Opioid
gamma-Aminobutyric Acid

Figure

• 

  Fig. 1 Time course of hindpaw withdrawal responses to von Frey filament pressure after (A) incisional surgery or (B) carrageenan injection. Data are presented as withdrawal thresholds. Each line represents the mean ± SEM of data from five or six rats. BL: baseline withdrawal threshold measured prior to paw incision or carrageenan injection. Significant differences between data from the ipsilateral and contralateral sites are indicated, *P < 0.05, †P < 0.01, ‡P < 0.001.

• 

  Fig. 2 Expression levels of mRNAs encoding γ-aminobutyric acid (GABA) and opioid receptors in the left dorsal horn of the spinal cord in naïve, incised, and carrageenan-injected rats. Data are presented as relative values of GABA and opioid receptors to that of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Each bar represents the mean ± SEM of data from four rats. Reverse transcriptase polymerase chain reaction detected mRNA bands corresponding to the GABA and opioid receptors in naïve animals. (A) Quantitative analysis indicated that incisional surgery reduced the levels of expression of mRNAs encoding GABA receptors, but not of those encoding opioid receptors. (B) Carrageenan injection increased expression of opioid receptor mRNAs, but not those of GABA receptor mRNAs. *P < 0.05, †P < 0.01, ‡P < 0.001 compared to naïve rats.

• 

  Fig. 3 (A) The time effect and (B) dose-response of intrathecal curcumin on the hindpaw withdrawal response to von Frey filament pressure applied after incisional surgery. Data are presented as withdrawal thresholds or percentages of maximal possible effects (%MPEs). Each line represents the mean ± SEM of data from five-to-seven rats. Control data were obtained 2 h after incision. Curcumin was administered immediately after measurement of control post-incisional thresholds. BL: baseline withdrawal threshold measured before paw incision. Curcumin produced dose-dependent increases in withdrawal thresholds of injured paws after incision. *P < 0.05, †P < 0.01 compared to vehicle (dimethylsulfoxide, DMSO) alone.

• 

  Fig. 4 (A) The time effect of and (B) dose-response to intrathecal curcumin, assessed by measuring hindpaw withdrawal responses to von Frey filament pressure applied after carrageenan injection. Data are presented as withdrawal thresholds or percentages of maximal possible effects (%MPEs). Each line represents the mean ± SEM of data from five-to-seven rats. Control data were obtained 4 h after carrageenan injection. Curcumin was administered immediately after measurement of control post-injectional thresholds. BL: baseline withdrawal threshold measured before carrageenan injection. Curcumin caused a dose-dependent increase in the withdrawal thresholds of injected paws after carrageenan injection. *P < 0.05 compared to vehicle (dimethylsulfoxide, DMSO) alone.

• 

  Fig. 5 (A) The effects of intrathecal bicuculline (a γ-aminobutyric acid [GABA]-A receptor antagonist, 0.3 μg), and saclofen (a GABA-B receptor antagonist, 100 μg), on the antinociceptive effects of intrathecal curcumin (100 μg) in rats experiencing postoperative pain; and (B) the effects of intrathecal curcumin (100 μg) on the expression levels of mRNAs encoding GABA receptors in the left dorsal horn of the spinal cord in incised rats. Data are presented as percentages of maximal possible effects (%MPEs) or the relative values of GABA receptor levels to that of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Each bar represents the mean ± SEM of data from four-to-six rats. DMSO: dimethylsulfoxide, CUR: curcumin, BI: Bicuculline, SA: saclofen. (A) Both bicuculline and saclofen reversed the effects of curcumin, †P < 0.01, ‡P < 0.001 compared with curcumin. (B) Intrathecal curcumin increased the expression levels of mRNAs encoding GABA receptors, †P < 0.01, ‡P < 0.001 compared with the incised group.

• 

  Fig. 6 (A) The effects of intrathecal CTOP (a μ opioid receptor antagonist, 15 μg), naltrindole (a δ opioid receptor antagonist, 10 g), and GNTI (a κ opioid receptor antagonist, 50 μg) on the antinociceptive effects of intrathecal curcumin (100 μg) on postoperative pain, and (B) the effects of intrathecal curcumin (100 μg) on the expression levels of mRNAs encoding opioid receptors in the left dorsal horn of the spinal cord in incised rats. Data are presented as percentages of maximal possible effects (%MPEs) or the relative expression levels of opioid receptors compared to that of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Each bar represents the mean ± SEM of data from four-to-six rats. DMSO: dimethylsulfoxide, CUR: curcumin, NALT: naltrindole. (A) CTOP, naltrindole and GNTI reversed the effects of curcumin, ‡P < 0.001 compared with curcumin. (B) Intrathecal curcumin only marginally affected the expression levels of mRNAs encoding opioid receptors in the incised group.

• 

  Fig. 7 (A) The effects of intrathecal CTOP (a μ opioid receptor antagonist, 15 μg), naltrindole (a δ opioid receptor antagonist, 10 μg) and GNTI (a κ opioid receptor antagonist, 50 μg) on the antinociceptive effects of intrathecal curcumin (300 μg) on inflammatory pain, and (B) the effects of intrathecal curcumin (300 μg) on the expression levels of mRNAs encoding opioid receptors in the left dorsal horn of the spinal cord in carrageenan-injected rats. Data are presented as percentages of maximal possible effects (%MPEs) or the relative expression levels of opioid receptors compared to that of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Each bar represents the mean ± SEM of data from four-to-six rats. DMSO: dimethylsulfoxide, CUR: curcumin, NALT: naltrindole. (A) All of CTOP, naltrindole, and GNTI reversed the effects of curcumin. ‡P < 0.001 compared with curcumin. (B) Intrathecal curcumin decreased the expression levels of mRNAs encoding all three opioid receptors, *P < 0.05, †P < 0.01, compared to the inflammatory group.

• 

  Fig. 8 (A) The effects of intrathecal bicuculline (a γ-aminobutyric acid [GABA]-A receptor antagonist, 0.3 μg) and saclofen (a GABA-B receptor antagonist, 100 μg) on the antinociceptive effects of intrathecal curcumin (300 μg) on inflammatory pain, and (B) the effects of intrathecal curcumin (300 μg) on the expression levels of mRNAs encoding GABA receptors in the left dorsal horn of the spinal cord in carrageenan-injected rats. Data are presented as percentages of maximal possible effects (%MPEs) or the relative expression levels of GABA receptors to that of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Each bar represents the mean ± SEM of data from four-to-six rats. DMSO: dimethylsulfoxide, CUR: curcumin, BI: Bicuculline, SA: saclofen. (A) Both bicuculline and saclofen reversed the effects of curcumin. †P < 0.01 compared with curcumin. (B) Intrathecal curcumin only marginally affected the expression levels of mRNAs encoding GABA receptors in the inflammatory group.

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