Korean J Pain.  2022 Jan;35(1):33-42. 10.3344/kjp.2022.35.1.33.

Anti-nociceptive and anti-inflammatory activities of the essential oil isolated from Cupressus arizonica Greene fruits

• Fakhri S
• Jafarian S
• Majnooni MB
• Farzaei MH
• Mohammadi-Noori E
• Khan H

• KMID: 2524645
• DOI: http://doi.org/10.3344/kjp.2022.35.1.33

Abstract

Background
Cupressus arizonica Greene is a coniferous tree with great importance in fragrance and pharmaceutical industries. Essential oils from C. arizonica (EC) have shown potential antioxidant, and anti-microbial activities. This study aimed at investigating the anti-nociceptive and anti-inflammatory effects/mechanisms of EC.
Methods
The EC was evaluated for anti-nociceptive and anti-inflammatory activities on male Wistar rats using a formalin test and carrageenan-induced paw edema, respectively. Also, we pre-treated some of the animals with naloxone and flumazenil in the formalin test to find out the possible contributions of opioid and benzodiazepine receptors to EC anti-nociceptive effects. Finally, gas chromatography/mass spectrometry (GC/MS) analysis was used to identify the EC’s constituents.
Results
EC in intraperitoneal doses of 0.5 and 1 g/kg significantly decrease the nociceptive responses in both early and late phases of the formalin test. From a mechanistic point of view, flumazenil administration 20 minutes before the most effective dose of EC (1 g/kg) showed a meaningful reduction in the associated antinociceptive responses during the early and late phases of the formalin test. Naloxone also reduced the anti-nociceptive role of EC in the late phase. Furthermore, EC at the doses of 1, 0.5, and 0.25 g/kg significantly reduced paw edema from 0.5 hours after carrageenan injection to 4 hours. GC/MS analysis showed that isolated EC is a monoterpene-rich oil with the major presence of α-pinene (71.92%), myrcene (6.37%), δ-3-carene (4.68%), β-pinene (3.71%), and limonene (3.34%).
Conclusions
EC showed potent anti-nociceptive and anti-inflammatory activities with the relative involvement of opioid and benzodiazepine receptors.

Keyword

Analgesics; Opioid; Anti-Inflammatory Agents; Antioxidants; Cupressus; Gas Chromatography-Mass Spectrometry; Inflammation; Oils; Volatile; Pain Measurement; Rats; Wistar; Receptors; GABA-A

Figure

• 

  Fig. 1 Anti-nociceptive activity of Cupressus arizonica crushed fruits EC in the early (A) and late (B) phases of formalin test. The error bars indicate standard error of mean. Control: negative control group, Diclofenac: positive control group, S1: 1 g/kg EC, S2: 0.5 g/kg EC, EC: essential oil of C. arizonica. ***P < 0.001 vs. Control. +++P < 0.001 vs. Diclofenac.

• 

  Fig. 2 Comparison of the effects of opioid (naloxone) and benzodiazepine (flumazenil) antagonists on the reduction of EC anti-nociceptive activity in the early (A) and late (B) phases of formalin test. From the left: Control: negative control group, Diclofenac: positive control group, Naloxone: the group which received naloxone and not EC, Flumazenil: the group which received flumazenil and not EC. The remaining three columns of the right-hand side underlined by “Cupressus arizonica (g/kg)” indicate the groups treated with 1 g/kg of C. arizonica EC following pre-treatment with the stated antagonists (naloxone, flumazenil) or without any pre-treatment (S1). The error bars indicate standard error of mean. EC: essential oil of C. arizonica. **P < 0.01. ***P < 0.001 vs. Control. ++P < 0.01. +++P < 0.001 vs. Diclofenac. ##P < 0.01. ###P < 0.001 vs. Flumazenil. @P < 0.05 vs. Naloxone.

• 

  Fig. 3 Anti-inflammatory effects of different doses of EC as evidenced by decreased paw edema at different time intervals including 0.5 (B), 1 (C), 2 (D) and 4 (E) hours post carrageenan injection. (A) indicates the time when carrageenan was sub-plantarly injected. The error bars indicate standard error of mean. EC: essential oils of Cupressus arizonica, Control: negative control group, Diclofenac: positive control group, S1: 1 g/kg EC, S2: 0.5 g/kg EC, S3: 0.25 g/kg EC. *P < 0.05. **P < 0.01. ***P < 0.001 vs. Control.

• 

  Fig. 4 GC/MS analysis of EC isolated from Cupressus arizonica Greene fruit crushed fruits. GC/MS: gas chromatography/mass spectrometry, EC: essential oil of C. arizonica.

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