Korean J Pain.  2021 Apr;34(2):185-192. 10.3344/kjp.2021.34.2.185.

Effect of pregabalin on nociceptive thresholds and immune responses in a mouse model of incisional pain

• Park JH
• Cho SH
• Kim R
• Na SH
• Kang Es
• Yeom My
• Jang Y

• KMID: 2514418
• DOI: http://doi.org/10.3344/kjp.2021.34.2.185

Abstract

Background
It is known that some analgesics as well as pain can affect the immune system. The aim of this study was to investigate the analgesic effect and immunomodulation of pregabalin (PGB) in a mouse incisional pain model.
Methods
A postoperative pain model was induced by hind paw plantar incision in male BALB/c mice. Mice were randomly divided into four groups (n = 8): a salinetreated incision (incision), PGB-treated incision (PGB-incision), sham controls without incision or drug treatment (control), and a PGB-treated control (PGB-control). In the PGB treated groups, PGB was administered intraperitoneally (IP) 30 minutes before and 1 hour after the plantar incision. Changes of the mechanical nociceptive thresholds following incision were investigated. Mice were euthanized for spleen harvesting 12 hours after the plantar incision, and natural killer (NK) cytotoxicity to YAC 1 cells and lymphocyte proliferation responses to phytohemagglutinin were compared among these four groups.
Results
Mechanical nociceptive thresholds were decreased after plantar incision and IP PGB administration recovered these decreased mechanical nociceptive thresholds (P < 0.001). NK activity was increased by foot incision, but NK activity in the PGB-incision group was significantly lower than that in the Incision group (P < 0.001). Incisional pain increased splenic lymphocyte proliferation, but PGB did not alter this response.
Conclusions
Incisional pain alters cell immunity of the spleen in BALB/c mice. PGB showed antinocieptive effect on mouse incisional pain and attenuates the activation of NK cells in this painful condition. These results suggest that PGB treatment prevents increases in pain induced NK cell activity.

Keyword

Analgesics; Immunity; Cellular; Immunomodulation; Killer Cells; Natural; Mice; Pain; Postoperative; Pregabalin; Spleen

Figure

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  Fig. 1 Experimental procedure. At the beginning of the experiment, initial paw withdrawal threshold were assessed for mechanical nociceptive test (baseline). In the pregabalin (PGB)-incision and PGB-control groups, PGB in saline was administered intraperitoneally, at 30 mg/kg, in a volume of 0.1 mL/10 g. In the incision group, an identical volume of saline was injected intraperitoneally. Thirty minutes subsequent to the initial drug injection, plantar incision or anesthesia was applied in accordance with group. One hour following hind paw incision, a second intraperitoneal injection was performed at the same dosage. Mice were sacrificed by cervical dislocation 12 hours following plantar incision, and natural killer cytotoxicity and lymphocyte proliferation were measured. The figure shows the whole experimental process using group 3 as an example.

• 

  Fig. 2 Time course of paw withdrawal threshold (PWT) after plantar incision in mice (n = 8 per group). Foot incision resulted in the development of marked mechanical allodynia until the day after the incision in the incision group (saline + incision). Intraperitoneal injection of pregabalin (PGB) at 30 mg/kg significantly reduced mechanical allodynia in the PGB-incision group (PGB + incision). There were no significant differences in the PWT between the control and PGB-incision groups throughout the experiment. In the control and PGB-incision groups, there were no significant changes in the PWT compared with the baseline PWT. Data represent means ± standard deviation. aP < 0.001 vs. the control and PGB-incision groups. bP < 0.001 vs. baseline.

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  Fig. 3 Effects of perioperative pregabalin (PGB) treatment on natural killer (NK) cell activity (% specific lysis) on the day after hind paw incision in mice (n = 8 per group). At various E:T ratios (80:1, 40:1, 20:1), the tumoricidal activities of NK cells (effector cells, E) against NK sensitive YAC-1 lymphoma cells (target cell, T) were calculated from lactate dehydrogenase release. NK cell activity was significantly increased in the incision group (saline + incision). In the PGB-incision group (PGB + incision), perioperative intraperitoneal PGB treatment significantly reduced NK activity compared with the incision group, but NK activity in the PGB-incision group remained higher than in the control group (P < 0.001). Data represent means ± standard deviation. aP < 0.001 vs. the PGB-incision, control, and PGB-control groups. bP < 0.001 vs. the control and PGB-control groups.

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  Fig. 4 Effects of perioperative pregabalin (PGB) treatment on splenocyte proliferation in mice the day following foot incision (n = 8 per group). Proliferation in response to 5 µg of phytohemagglutinin mitogenic stimulation was determined as the percentage changes in the stimulation index. Incision-induced pain, in both the incision and PGB-incision groups, increased splenic lymphocyte proliferation to a significantly greater degree compared with the control group. However, PGB pretreatment in the PGB-control and PGB-incision groups did not induce greater changes in splenocyte proliferation compared with the control and incision groups, respectively. Data represent means ± standard deviation. aP < 0.05 vs. control.

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  Fig. 5 Change in plasma pregabalin (PGB) concentration over time. In this auxiliary experiment, six mice underwent plantar incision and preoperative and postoperative PGB treatment, and blood was taken 2, 6, and 12 hours after the first PGB injection. Plasma PGB concentrations were measured by using high-performance liquid chromatography/mass spectrometry. Twelve hours after the first injection, which is when spleens were harvested for immune response analyses, the PGB concentration was close to zero (0.013 ± 0.003 µg/mL). Data represent means ± standard deviation.

Reference

1. Kennedy BC, Hall GM. 1999; Neuroendocrine and inflammatory aspects of surgery: do they affect outcome? Acta Anaesthesiol Belg. 50:205–9. PMID: 10603996.

2. Kiecolt-Glaser JK, Page GG, Marucha PT, MacCallum RC, Glaser R. 1998; Psychological influences on surgical recovery. Perspectives from psychoneuroimmunology. Am Psychol. 53:1209–18. DOI: 10.1037/0003-066X.53.11.1209. PMID: 9830373.

3. Kehlet H, Holte K. 2001; Effect of postoperative analgesia on surgical outcome. Br J Anaesth. 87:62–72. DOI: 10.1093/bja/87.1.62. PMID: 11460814.

4. Tsai YC, Won SJ, Lin MT. 2000; Effects of morphine on immune response in rats with sciatic constriction injury. Pain. 88:155–60. DOI: 10.1016/S0304-3959(00)00323-7. PMID: 11050370.

5. Beilin B, Shavit Y, Trabekin E, Mordashev B, Mayburd E, Zeidel A, et al. 2003; The effects of postoperative pain management on immune response to surgery. Anesth Analg. 97:822–7. DOI: 10.1213/01.ANE.0000078586.82810.3B. PMID: 12933409.

6. Sacerdote P, Bianchi M, Manfredi B, Panerai AE. 1997; Effects of tramadol on immune responses and nociceptive thresholds in mice. Pain. 72:325–30. DOI: 10.1016/S0304-3959(97)00055-9. PMID: 9313273.

7. Jang Y, Song HK, Yeom MY, Jeong DC. 2012; The immunomodulatory effect of pregabalin on spleen cells in neuropathic mice. Anesth Analg. 115:830–6. DOI: 10.1213/ANE.0b013e31825d2ad1. PMID: 22729962.

8. van Seventer R, Bach FW, Toth CC, Serpell M, Temple J, Murphy TK, et al. 2010; Pregabalin in the treatment of post-traumatic peripheral neuropathic pain: a randomized double-blind trial. Eur J Neurol. 17:1082–9. DOI: 10.1111/j.1468-1331.2010.02979.x. PMID: 20236172.

9. Field MJ, Holloman EF, McCleary S, Hughes J, Singh L. 1997; Evaluation of gabapentin and S-(+)-3-isobutylgaba in a rat model of postoperative pain. J Pharmacol Exp Ther. 282:1242–6. PMID: 9316831.

10. Zhang J, Ho KY, Wang Y. 2011; Efficacy of pregabalin in acute postoperative pain: a meta-analysis. Br J Anaesth. 106:454–62. DOI: 10.1093/bja/aer027. PMID: 21357616.

11. Pogatzki EM, Raja SN. 2003; A mouse model of incisional pain. Anesthesiology. 99:1023–7. DOI: 10.1097/00000542-200310000-00041. PMID: 14508341.

12. Pogatzki EM, Vandermeulen EP, Brennan TJ. 2002; Effect of plantar local anesthetic injection on dorsal horn neuron activity and pain behaviors caused by incision. Pain. 97:151–61. DOI: 10.1016/S0304-3959(02)00014-3. PMID: 12031788.

13. Hegarty DA, Shorten GD. 2011; A randomised, placebo-controlled trial of the effects of preoperative pregabalin on pain intensity and opioid consumption following lumbar discectomy. Korean J Pain. 24:22–30. DOI: 10.3344/kjp.2011.24.1.22. PMID: 21390175. PMCID: PMC3049973.

14. Lam DMH, Choi SW, Wong SSC, Irwin MG, Cheung CW. 2015; Efficacy of pregabalin in acute postoperative pain under different surgical categories: a meta-analysis. Medicine (Baltimore). 94:e1944. DOI: 10.1097/MD.0000000000001944. PMID: 26579802. PMCID: PMC4652811.

15. Chesler EJ, Ritchie J, Kokayeff A, Lariviere WR, Wilson SG, Mogil JS. 2003; Genotype-dependence of gabapentin and pregabalin sensitivity: the pharmacogenetic mediation of analgesia is specific to the type of pain being inhibited. Pain. 106:325–35. DOI: 10.1016/S0304-3959(03)00330-0. PMID: 14659515.

16. Field MJ, Oles RJ, Lewis AS, McCleary S, Hughes J, Singh L. 1997; Gabapentin (neurontin) and S-(+)-3-isobutylgaba represent a novel class of selective antihyperalgesic agents. Br J Pharmacol. 121:1513–22. DOI: 10.1038/sj.bjp.0701320. PMID: 9283683. PMCID: PMC1564875.

17. Takeuchi Y, Takasu K, Ono H, Tanabe M. 2007; Pregabalin, S-(+)-3-isobutylgaba, activates the descending noradrenergic system to alleviate neuropathic pain in the mouse partial sciatic nerve ligation model. Neuropharmacology. 53:842–53. DOI: 10.1016/j.neuropharm.2007.08.013. PMID: 17889907.

18. Sacerdote P, Bianchi M, Gaspani L, Manfredi B, Maucione A, Terno G, et al. 2000; The effects of tramadol and morphine on immune responses and pain after surgery in cancer patients. Anesth Analg. 90:1411–4. DOI: 10.1097/00000539-200006000-00028. PMID: 10825330.

19. Sharify A, Mahmoudi M, Izad MH, Hosseini MJ, Sharify M. 2007; Effect of acute pain on splenic NK cell activity, lymphocyte proliferation and cytokine production activities. Immunopharmacol Immunotoxicol. 29:465–76. DOI: 10.1080/08923970701619877. PMID: 18075858.

20. Eisenstein TK, Hilburger ME. 1998; Opioid modulation of immune responses: effects on phagocyte and lymphoid cell populations. J Neuroimmunol. 83:36–44. DOI: 10.1016/S0165-5728(97)00219-1. PMID: 9610671.

21. Ren K, Dubner R. 2010; Interactions between the immune and nervous systems in pain. Nat Med. 16:1267–76. DOI: 10.1038/nm.2234. PMID: 20948535. PMCID: PMC3077564.

22. Zajączkowska R, Leppert W, Mika J, Kocot-Kępska M, Woroń J, Wrzosek A, et al. 2018; Perioperative immunosuppression and risk of cancer progression: the impact of opioids on pain management. Pain Res Manag. 2018:9293704. DOI: 10.1155/2018/9293704. PMID: 30327708. PMCID: PMC6169211.

23. Peterson PK, Molitor TW, Chao CC. 1993; Mechanisms of morphine-induced immunomodulation. Biochem Pharmacol. 46:343–8. DOI: 10.1016/0006-2952(93)90508-T. PMID: 8394079.

24. Lysle DT, Coussons ME, Watts VJ, Bennett EH, Dykstra LA. 1993; Morphine-induced alterations of immune status: dose dependency, compartment specificity and antagonism by naltrexone. J Pharmacol Exp Ther. 265:1071–8. PMID: 7685383.

25. Mössner R, Lesch KP. 1998; Role of serotonin in the immune system and in neuroimmune interactions. Brain Behav Immun. 12:249–71. DOI: 10.1006/brbi.1998.0532. PMID: 10080856.

26. Madden KS, Moynihan JA, Brenner GJ, Felten SY, Felten DL, Livnat S. 1994; Sympathetic nervous system modulation of the immune system. III. Alterations in T and B cell proliferation and differentiation in vitro following chemical sympathectomy. J Neuroimmunol. 49:77–87. DOI: 10.1016/0165-5728(94)90183-X. PMID: 8294564.

27. Tanabe M, Takasu K, Takeuchi Y, Ono H. 2008; Pain relief by gabapentin and pregabalin via supraspinal mechanisms after peripheral nerve injury. J Neurosci Res. 86:3258–64. DOI: 10.1002/jnr.21786. PMID: 18655202.