Korean J Pain.  2021 Jan;34(1):35-46. 10.3344/kjp.2021.34.1.35.

Antinociceptive effects of oleuropein in experimental models of neuropathic pain in male rats

• Chen H
• Ma D
• Zhang H
• Tang Y
• Wang J
• Li R
• Wen W
• Zhang Y

• KMID: 2510310
• DOI: http://doi.org/10.3344/kjp.2021.34.1.35

Abstract

Background
The present investigation explored the therapeutic actions of oleuropein along with the possible signaling pathway involved in attenuating neuropathic pain in chronic constriction injury (CCI) and vincristine-induced neuropathic pain in male rats.
Methods
Four loose ligatures were placed around the sciatic nerve to induce CCI, and vincristine (50 μg/kg) was injected for 10 days to develop neuropathic pain. The development of cold allodynia, mechanical allodynia, and mechanical hyperalgesia was assessed using different pain-related behavioral tests. The levels of H₂S, cystathionine-γ-lyase (CSE), cystathionine-β-synthase (CBS), orexin, and nuclear factor erythroid-2-related factor 2 (Nrf2) were measured in the sciatic nerve.
Results
Treatment with oleuropein for 14 days led to significant amelioration of behavioral manifestations of neuropathic pain in two pain models. Moreover, oleuropein restored both CCI and vincristine-induced decreases in H₂S, CSE, CBS, orexin, and Nrf2 levels. Co-administration of suvorexant, an orexin receptor antagonist, significantly counteracted the pain-attenuating actions of oleuropein and Nrf2 levels without modulating H₂S, CSE and CBS.
Conclusions
Oleuropein has therapeutic potential to attenuate the pain manifestations in CCI and vincristine-induced neuropathic pain, possibly by restoring the CSE, CBS, and H₂S, which may subsequently increase the expression of orexin and Nrf2 to ameliorate behavioral manifestations of pain.

Keyword

Hydrogen Sulfide; Hyperalgesia; Neuralgia; Neuropeptides; NF-E2-Related Factor 2; Nociception; Orexins; Orexin Receptor Antagonists; Polyphenols; Vincristine

Figure

• 

  Fig. 1 Diagrammatic representation of experimental protocol in (A) chronic constriction injury (CCI) and (B) vincristine model. OLE: oleuropein, Suv: suvorexant, p.o.: per oral, i.p.: intraperitoneal, DMSO: dimethyl sulfoxide, VIN: vincristine-induced neuropathic pain.

• 

  Fig. 2 Effect of different interventions on the cold allodynia in the acetone drop test in chronic constriction injury (CCI) and vincristine models. Values are in mean ± standard deviation. N = 10, F(14, 135) = 342.5. OLE: oleuropein, Suv: suvorexant, DMSO: dimethyl sulfoxide, VIN: vincristine-induced neuropathic pain. aP < 0.05 vs. sham for CCI, bP < 0.05 vs. CCI, cP < 0.05 vs. OLE (20 mg/kg) in CCI, dP < 0.05 vs. sham for VIN, eP < 0.05 VIN, fP < 0.05 OLE (20 mg/kg) in VIN.

• 

  Fig. 3 Effect of different interventions on mechanical allodynia in the Von Frey filament test in chronic constriction injury (CCI) and vincristine models. Values are in mean ± standard deviation. N = 10, F(14, 135) = 372.7. OLE: oleuropein, Suv: suvorexant, DMSO: dimethyl sulfoxide, VIN: vincristine-induced neuropathic pain. aP < 0.05 vs. sham for CCI, bP < 0.05 vs. CCI, cP < 0.05 vs. OLE (20 mg/kg) in CCI, dP < 0.05 vs. sham for VIN, eP < 0.05 VIN, fP < 0.05 OLE (20 mg/kg) in VIN.

• 

  Fig. 4 Effect of different interventions on mechanical hyperalgesia in the pin-prick test in chronic constriction injury (CCI) and vincristine models. Values are in mean ± standard deviation. N = 10, F(14, 135) = 313.8. OLE: oleuropein, Suv: suvorexant, DMSO: dimethyl sulfoxide, VIN: vincristine-induced neuropathic pain. aP < 0.05 vs. sham for CCI, bP < 0.05 vs. CCI, cP < 0.05 vs. OLE (20 mg/kg) in CCI, dP < 0.05 vs. sham for VIN, eP < 0.05 VIN, fP < 0.05 OLE (20 mg/kg) in VIN.

• 

  Fig. 5 Effect of different interventions on H2S levels in the sciatic nerve in chronic constriction injury (CCI) and vincristine models. Values are in mean ± standard deviation. N = 10, F(14, 135) = 140.5. OLE: oleuropein, Suv: suvorexant, DMSO: dimethyl sulfoxide, VIN: vincristine-induced neuropathic pain. aP < 0.05 vs. sham for CCI, bP < 0.05 vs. CCI, cP < 0.05 vs. sham for VIN, dP < 0.05 VIN.

• 

  Fig. 6 Effect of different interventions on cystathionine-γ-lyase levels in the sciatic nerve in chronic constriction injury (CCI) and vincristine models. Values are in mean ± standard deviation. N = 10, F(14, 135) = 120.2. OLE: oleuropein, Suv: suvorexant, DMSO: dimethyl sulfoxide, VIN: vincristine-induced neuropathic pain. aP < 0.05 vs. sham for CCI, bP < 0.05 vs. CCI, cP < 0.05 vs. sham for VIN, dP < 0.05 VIN.

• 

  Fig. 7 Effect of different interventions in cystathionine-β-synthase levels in the sciatic nerve in chronic constriction injury (CCI) and vincristine models. Values are in mean ± standard deviation. N = 10, F(14, 135) = 132.7. OLE: oleuropein, Suv: suvorexant, DMSO: dimethyl sulfoxide, VIN: vincristine-induced neuropathic pain. aP < 0.05 vs. sham for CCI, bP < 0.05 vs. CCI, cP < 0.05 vs. sham for VIN, dP < 0.05 VIN.

• 

  Fig. 8 Effect of different interventions on orexin levels in the sciatic nerve in chronic constriction injury (CCI) and vincristine models. Values are in mean ± standard deviation. N = 10, F(14, 135) = 154.4. OLE: oleuropein, Suv: suvorexant, DMSO: dimethyl sulfoxide, VIN: vincristine-induced neuropathic pain. aP < 0.05 vs. sham for CCI, bP < 0.05 vs. CCI, cP < 0.05 vs. sham for VIN, dP < 0.05 VIN.

• 

  Fig. 9 Effect of different interventions on nuclear factor erythroid-2-related factor 2 levels in the sciatic nerve in chronic constriction injury (CCI) and vincristine models. Values are in mean ± standard deviation. N = 10, F(14, 135) = 144.2. OLE: oleuropein, Suv: suvorexant, DMSO: dimethyl sulfoxide, VIN: vincristine-induced neuropathic pain. aP < 0.05 vs. sham for CCI, bP < 0.05 vs. CCI, cP < 0.05 vs. OLE (20 mg/kg) in CCI, dP < 0.05 vs. sham for VIN, eP < 0.05 VIN, fP < 0.05 OLE (20 mg/kg) in VIN.

Reference

1. Zilliox LA. 2017; Neuropathic Pain. Continuum (Minneap Minn). 23(2, Selected Topics in Outpatient Neurology):512–32. DOI: 10.1212/CON.0000000000000462. PMID: 28375916.

2. Badr AM, Attia HA, Al-Rasheed N. 2020; Oleuropein reverses repeated corticosterone-induced depressive-like behavior in mice: evidence of modulating effect on biogenic amines. Sci Rep. 10:3336. DOI: 10.1038/s41598-020-60026-1. PMID: 32094406. PMCID: PMC7040186.

3. Mohammad-Beigi H, Aliakbari F, Sahin C, Lomax C, Tawfike A, Schafer NP, et al. 2019; Oleuropein derivatives from olive fruit extracts reduce α-synuclein fibrillation and oligomer toxicity. J Biol Chem. 294:4215–32. DOI: 10.1074/jbc.RA118.005723. PMID: 30655291. PMCID: PMC6422090.

4. Fki I, Sayadi S, Mahmoudi A, Daoued I, Marrekchi R, Ghorbel H. 2020; Comparative study on beneficial effects of hydroxytyrosol- and oleuropein-rich olive leaf extracts on high-fat diet-induced lipid metabolism disturbance and liver injury in rats. Biomed Res Int. 2020:1315202. DOI: 10.1155/2020/1315202. PMID: 31998777. PMCID: PMC6970490.

5. Czerwińska ME, Gąsińska E, Leśniak A, Krawczyk P, Kiss AK, Naruszewicz M, et al. 2018; Inhibitory effect of Ligustrum vulgare leaf extract on the development of neuropathic pain in a streptozotocin-induced rat model of diabetes. Phytomedicine. 49:75–82. DOI: 10.1016/j.phymed.2018.06.006. PMID: 30217264.

6. Mao X, Xia B, Zheng M, Zhou Z. 2019; Assessment of the anti-inflammatory, analgesic and sedative effects of oleuropein from Olea europaea L. Cell Mol Biol (Noisy-le-grand). 65:52–5. DOI: 10.14715/cmb/2019.65.1.9. PMID: 30782294.

7. Zare L, Esmaeili-Mahani S, Abbasnejad M, Rasoulian B, Sheibani V, Sahraei H, et al. 2012; Oleuropein, chief constituent of olive leaf extract, prevents the development of morphine antinociceptive tolerance through inhibition of morphine-induced L-type calcium channel overexpression. Phytother Res. 26:1731–7. DOI: 10.1002/ptr.4634. PMID: 22422486.

8. Linden DR. 2014; Hydrogen sulfide signaling in the gastrointestinal tract. Antioxid Redox Signal. 20:818–30. DOI: 10.1089/ars.2013.5312. PMID: 23582008. PMCID: PMC3910452.

9. Wu D, Hu Q, Zhu D. 2018; An update on hydrogen sulfide and nitric oxide interactions in the cardiovascular system. Oxid Med Cell Longev. 2018:4579140. DOI: 10.1155/2018/4579140. PMID: 30271527. PMCID: PMC6151216.

10. Sheibani L, Lechuga TJ, Zhang H, Hameed A, Wing DA, Kumar S, et al. 2017; Augmented H₂S production via cystathionine-beta-synthase upregulation plays a role in pregnancy-associated uterine vasodilation. Biol Reprod. 96:664–72. DOI: 10.1095/biolreprod.116.143834. PMID: 28339573. PMCID: PMC6366540.

11. Lin JQ, Luo HQ, Lin CZ, Chen JZ, Lin XZ. 2014; Sodium hydrosulfide relieves neuropathic pain in chronic constriction injured rats. Evid Based Complement Alternat Med. 2014:514898. DOI: 10.1155/2014/514898. PMID: 25506383. PMCID: PMC4260443.

12. Chen H, Xie K, Chen Y, Wang Y, Wang Y, Lian N, et al. 2019; Nrf2/HO-1 signaling pathway participated in the protection of hydrogen sulfide on neuropathic pain in rats. Int Immunopharmacol. 75:105746. DOI: 10.1016/j.intimp.2019.105746. PMID: 31325725.

13. Lucarini E, Micheli L, Trallori E, Citi V, Martelli A, Testai L, et al. 2018; Effect of glucoraphanin and sulforaphane against chemotherapy-induced neuropathic pain: Kv7 potassium channels modulation by H2 S release in vivo. Phytother Res. 32:2226–34. DOI: 10.1002/ptr.6159. PMID: 30069944.

14. Mieda M. 2017; The roles of orexins in sleep/wake regulation. Neurosci Res. 118:56–65. DOI: 10.1016/j.neures.2017.03.015. PMID: 28526554.

15. Suyama H, Kawamoto M, Shiraishi S, Gaus S, Kajiyama S, Yuge O. 2004; Analgesic effect of intrathecal administration of orexin on neuropathic pain in rats. In Vivo. 18:119–23. PMID: 15113038.

16. Toyama S, Shimoyama N, Shimoyama M. 2017; The analgesic effect of orexin-A in a murine model of chemotherapy-induced neuropathic pain. Neuropeptides. 61:95–100. DOI: 10.1016/j.npep.2016.12.007. PMID: 28041630.

17. Kajiyama S, Kawamoto M, Shiraishi S, Gaus S, Matsunaga A, Suyama H, et al. 2005; Spinal orexin-1 receptors mediate anti-hyperalgesic effects of intrathecally-administered orexins in diabetic neuropathic pain model rats. Brain Res. 1044:76–86. DOI: 10.1016/j.brainres.2005.03.007. PMID: 15862792.

18. Ma Q. 2013; Role of nrf2 in oxidative stress and toxicity. Annu Rev Pharmacol Toxicol. 53:401–26. DOI: 10.1146/annurev-pharmtox-011112-140320. PMID: 23294312. PMCID: PMC4680839.

19. Yetik-Anacak G, Sevin G, Ozzayım O, Dereli MV, Ahmed A. 2016; Hydrogen sulfide: a novel mechanism for the vascular protection by resveratrol under oxidative stress in mouse aorta. Vascul Pharmacol. 87:76–82. DOI: 10.1016/j.vph.2016.08.003. PMID: 27538867.

20. Wang Z, Yan Y, Wang Y, Tong F. 2019; The interaction between CSE/H₂S and the iNOS/NO-mediated resveratrol/poly(ethylene glycol)-poly(phenylalanine) complex alleviates intestinal ischemia/reperfusion injuries in diabetic rats. Biomed Pharmacother. 112:108736. DOI: 10.1016/j.biopha.2019.108736. PMID: 30970526.

21. Vitalone A, Di Sotto A, Mammola CL, Heyn R, Miglietta S, Mariani P, et al. 2017; Phytochemical analysis and effects on ingestive behaviour of a Caralluma fimbriata extract. Food Chem Toxicol. 108(Pt A):63–73. DOI: 10.1016/j.fct.2017.07.027. PMID: 28713048.

22. Guo C, Bi J, Li X, Lyu J, Liu X, Wu X, et al. 2021; Immunomodulation effects of polyphenols from thinned peach treated by different drying methods on RAW264.7 cells through the NF-κB and Nrf2 pathways. Food Chem. 340:127931. DOI: 10.1016/j.foodchem.2020.127931. PMID: 32871358.

23. Castejon ML, Sánchez-Hidalgo M, Aparicio-Soto M, Montoya T, Martín-LaCave I, Fernández-Bolaños JG, et al. 2019; Dietary oleuropein and its new acyl-derivate attenuate murine lupus nephritis through HO-1/Nrf2 activation and suppressing JAK/STAT, NF-κB, MAPK and NLRP3 inflammasome signaling pathways. J Nutr Biochem. 74:108229. DOI: 10.1016/j.jnutbio.2019.108229. PMID: 31698204.

24. Sanchez-Alavez M, Benedict J, Wills DN, Ehlers CL. 2019; Effect of suvorexant on event-related oscillations and EEG sleep in rats exposed to chronic intermittent ethanol vapor and protracted withdrawal. Sleep. 42:zsz020. DOI: 10.1093/sleep/zsz020. PMID: 30715515. PMCID: PMC6448295.

25. Janahmadi Z, Nekooeian AA, Moaref AR, Emamghoreishi M. 2017; Oleuropein attenuates the progression of heart failure in rats by antioxidant and antiinflammatory effects. Naunyn Schmiedebergs Arch Pharmacol. 390:245–52. DOI: 10.1007/s00210-016-1323-6. PMID: 27928616.

26. Bennett GJ, Xie YK. 1988; A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain. 33:87–107. DOI: 10.1016/0304-3959(88)90209-6. PMID: 2837713.

27. Ma L, Liu H, Chen G, Chen M, Wang L, Zhang X, et al. 2018; Sulfasalazine attenuates chronic constriction injury-induced neuroinflammation and mechanical hypersensitivity in rats. Neurosci Lett. 683:174–80. DOI: 10.1016/j.neulet.2018.07.042. PMID: 30075286.

28. Shibayama M, Kuniyoshi K, Suzuki T, Yamauchi K, Ohtori S, Takahashi K. 2014; The effects of locally injected triamcinolone on entrapment neuropathy in a rat chronic constriction injury model. J Hand Surg Am. 39:1714–21. DOI: 10.1016/j.jhsa.2014.05.026. PMID: 25017582.

29. Khangura RK, Bali A, Kaur G, Singh N, Jaggi AS. 2017; Neuropathic pain attenuating effects of perampanel in an experimental model of chronic constriction injury in rats. Biomed Pharmacother. 94:557–63. DOI: 10.1016/j.biopha.2017.07.137. PMID: 28780471.

30. Siau C, Bennett GJ. 2006; Dysregulation of cellular calcium homeostasis in chemotherapy-evoked painful peripheral neuropathy. Anesth Analg. 102:1485–90. DOI: 10.1213/01.ane.0000204318.35194.ed. PMID: 16632831. PMCID: PMC1805480.

31. Selawry OS, Hananian J. 1963; Vincristine treatment of cancer in children. JAMA. 183:741–6. DOI: 10.1001/jama.1963.03700090061010. PMID: 13992692.

32. Gilchrist L, Tanner L. 2016; Gait patterns in children with cancer and vincristine neuropathy. Pediatr Phys Ther. 28:16–22. DOI: 10.1097/PEP.0000000000000208. PMID: 27088678.

33. Vashistha B, Sharma A, Jain V. 2017; Ameliorative potential of ferulic acid in vincristine-induced painful neuropathy in rats: an evidence of behavioral and biochemical examination. Nutr Neurosci. 20:60–70. DOI: 10.1179/1476830514Y.0000000165. PMID: 25494651.

34. Choi Y, Yoon YW, Na HS, Kim SH, Chung JM. 1994; Behavioral signs of ongoing pain and cold allodynia in a rat model of neuropathic pain. Pain. 59:369–76. DOI: 10.1016/0304-3959(94)90023-X. PMID: 7708411.

35. Chaplan SR, Bach FW, Pogrel JW, Chung JM, Yaksh TL. 1994; Quantitative assessment of tactile allodynia in the rat paw. J Neurosci Methods. 53:55–63. DOI: 10.1016/0165-0270(94)90144-9. PMID: 7990513.

36. Wang G, Yang Y, Wang C, Huang J, Wang X, Liu Y, et al. 2020; Exploring the role and mechanisms of diallyl trisulfide and diallyl disulfide in chronic constriction-induced neuropathic pain in rats. Korean J Pain. 33:216–25. DOI: 10.3344/kjp.2020.33.3.216. PMID: 32606266. PMCID: PMC7336342.

37. Xie T, Zhang J, Kang Z, Liu F, Lin Z. 2020; miR-101 down-regulates mTOR expression and attenuates neuropathic pain in chronic constriction injury rat models. Neurosci Res. 158:30–6. DOI: 10.1016/j.neures.2019.09.002. PMID: 31526851.

38. Yamamoto S, Suzuki Y, Ono H, Kume K, Ohsawa M. 2016; N- and L-type calcium channels blocker cilnidipine ameliorates neuropathic pain. Eur J Pharmacol. 793:66–75. DOI: 10.1016/j.ejphar.2016.11.001. PMID: 27823932.

39. Shen X, Chakraborty S, Dugas TR, Kevil CG. 2014; Hydrogen sulfide measurement using sulfide dibimane: critical evaluation with electrospray ion trap mass spectrometry. Nitric Oxide. 41:97–104. DOI: 10.1016/j.niox.2014.06.002. PMID: 24932544. PMCID: PMC4413942.

40. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1951; Protein measurement with the Folin phenol reagent. J Biol Chem. 193:265–75. PMID: 14907713.

41. Kim YH, Choi YJ, Kang MK, Lee EJ, Kim DY, Oh H, et al. 2018; Oleuropein curtails pulmonary inflammation and tissue destruction in models of experimental asthma and emphysema. J Agric Food Chem. 66:7643–54. DOI: 10.1021/acs.jafc.8b01808. PMID: 29945446.

42. Lechuga TJ, Qi QR, Kim T, Magness RR, Chen DB. 2019; E2β stimulates ovine uterine artery endothelial cell H₂S production in vitro by estrogen receptor-dependent upregulation of cystathionine β-synthase and cystathionine γ-lyase expression†. Biol Reprod. 100:514–22. DOI: 10.1093/biolre/ioy207. PMID: 30277497. PMCID: PMC6378861.

43. Di Cesare Mannelli L, Lucarini E, Micheli L, Mosca I, Ambrosino P, Soldovieri MV, et al. 2017; Effects of natural and synthetic isothiocyanate-based H₂S-releasers against chemotherapy-induced neuropathic pain: role of Kv7 potassium channels. Neuropharmacology. 121:49–59. DOI: 10.1016/j.neuropharm.2017.04.029. PMID: 28431970.

44. Wu G, Ringkamp M, Hartke TV, Murinson BB, Campbell JN, Griffin JW, et al. 2001; Early onset of spontaneous activity in uninjured C-fiber nociceptors after injury to neighboring nerve fibers. J Neurosci. 21:RC140. DOI: 10.1523/JNEUROSCI.21-08-j0002.2001. PMCID: PMC6762537. PMID: 11306646.

45. Djouhri L, Koutsikou S, Fang X, McMullan S, Lawson SN. 2006; Spontaneous pain, both neuropathic and inflammatory, is related to frequency of spontaneous firing in intact C-fiber nociceptors. J Neurosci. 26:1281–92. DOI: 10.1523/JNEUROSCI.3388-05.2006. PMID: 16436616. PMCID: PMC6674571.

46. Meacham K, Shepherd A, Mohapatra DP, Haroutounian S. 2017; Neuropathic pain: central vs. peripheral mechanisms. Curr Pain Headache Rep. 21:28. DOI: 10.1007/s11916-017-0629-5. PMID: 28432601.

47. Razavi BM, Hosseinzadeh H. 2017; A review of the role of orexin system in pain modulation. Biomed Pharmacother. 90:187–93. DOI: 10.1016/j.biopha.2017.03.053. PMID: 28360013.

48. Sorge RE, Mapplebeck JC, Rosen S, Beggs S, Taves S, Alexander JK, et al. 2015; Different immune cells mediate mechanical pain hypersensitivity in male and female mice. Nat Neurosci. 18:1081–3. DOI: 10.1038/nn.4053. PMID: 26120961. PMCID: PMC4772157.

49. Castejón ML, Rosillo MÁ, Montoya T, González-Benjumea A, Fernández-Bolaños JG, Alarcón-de-la-Lastra C. 2017; Oleuropein down-regulated IL-1β-induced inflammation and oxidative stress in human synovial fibroblast cell line SW982. Food Funct. 8:1890–8. DOI: 10.1039/C7FO00210F. PMID: 28426090.

50. Dubey AK, Handu SS, Mediratta PK. 2015; Suvorexant: the first orexin receptor antagonist to treat insomnia. J Pharmacol Pharmacother. 6:118–21. DOI: 10.4103/0976-500X.155496. PMID: 25969666. PMCID: PMC4419247.