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Carvacrol Protects Against 6-Hydroxydopamine-Induced Neurotoxicity in In Vivo and In Vitro Models of Parkinson’s Disease

Abstract

Parkinson’s disease (PD) is a progressive neurodegenerative movement disorder characterized by selective loss of dopaminergic neurons that project from the substantia nigra pars compacta to the striatum. Evidence from human and animal studies has suggested that oxidative damage critically contributes to neuronal loss in PD. Carvacrol (CAR), a monoterpenic phenol, is the main constituents in the essential oil of many aromatic plants and possesses some properties including anti-inflammatory and anti-oxidant effects. In this study, in vitro and in vivo experiments were performed with the CAR in order to investigate its potential neuroprotective effects in models of PD. Post-treatment with CAR in vitro was found to protect rat adrenal pheochromocytoma PC12 cells from toxicity induced by 6-hydroxydopamine (6-OHDA) administration in a dose-dependent manner by (1) increasing cell viability and (2) reduction in intracellular reactive oxygen species, intracellular lipid peroxidation, and annexin-positive cells. In vivo, post-treatment with CAR (15 and 20 mg/kg) was protective against neurodegenerative phenotypes associated with systemic administration of 6-OHDA. Results indicated that CAR improved the locomotor activity, catalepsy, akinesia, bradykinesia, and motor coordination and reduced the apomorphine-caused rotation in 6-OHDA-stimulated rats. Increased level of reduced glutathione content and a decreased level of MDA (malondialdehyde) were observed in the 6-OHDA rats post-treated with CAR. These findings suggest that CAR exerts protective effects, possibly related to an anti-oxidation mechanism, in these in vitro and in vivo models of Parkinson’s disease.

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References

  1. Ahmad M, Yousuf S, Khan MB, Hoda MN, Ahmad AS, Ansari MA, Ishrat T, Agrawal AK, Islam F (2006) Attenuation by Nardostachys jatamansi of 6-hydroxydopamine-induced parkinsonism in rats: behavioral, neurochemical, and immunohistochemical studies. Pharmacol Biochem Behav 83(1):150–160

  2. Aminyavari S, Zahmatkesh M, Farahmandfar M, Khodagholi F, Dargahi L, Zarrindast M-R (2019) Protective role of Apelin-13 on amyloid β25–35-induced memory deficit; involvement of autophagy and apoptosis process. Prog Neuro-Psychopharmacol Biol Psychiatry 89:322–334

  3. Arunasree K (2010) Anti-proliferative effects of carvacrol on a human metastatic breast cancer cell line, MDA-MB 231. Phytomedicine 17(8–9):581–588

  4. Azizi Z, Ebrahimi S, Saadatfar E, Kamalinejad M, Majlessi N (2012) Cognitive-enhancing activity of thymol and carvacrol in two rat models of dementia. Behav Pharmacol 23(3):241–249

  5. Baluchnejadmojarad T, Hassanshahi J, Roghani M, Mansouri M, Raoufi S (2014) Protective effect of carvacrol in 6-hydroxydopamine hemi-parkinsonian rat model. JBCP 2(2):29–34

  6. Biocca S, Cattaneo A, Calissano P (1983) A macromolecular structure favouring microtubule assembly in NGF-differentiated pheochromocytoma cells (PC12). EMBO J 2(5):643–648

  7. Boskabady MH, Jafari Z, Pouraboli I (2011) The effect of carvacrol on muscarinic receptors of guinea-pig tracheal chains. Phytother Res 25(4):530–535. https://doi.org/10.1002/ptr.3290

  8. Broom L, Marinova-Mutafchieva L, Sadeghian M, Davis JB, Medhurst AD, Dexter DT (2011) Neuroprotection by the selective iNOS inhibitor GW274150 in a model of Parkinson disease. Free Radic Biol Med 50(5):633–640

  9. Chen W, Xu B, Xiao A, Liu L, Fang X, Liu R, Turlova E, Barszczyk A, Zhong X, Sun CLF, Britto LRG, Feng ZP, Sun HS (2015) TRPM7 inhibitor carvacrol protects brain from neonatal hypoxic-ischemic injury. Mol Brain 8(1):11

  10. Cho S, Choi Y, Park S, Park T (2012) Carvacrol prevents diet-induced obesity by modulating gene expressions involved in adipogenesis and inflammation in mice fed with high-fat diet. J Nutr Biochem 23(2):192–201

  11. Cohen G, Heikkila RE, Allis B, Cabbat F, Dembiec D, MacNamee D, Mytilineou C, Winston B (1976) Destruction of sympathetic nerve terminals by 6-hydroxydopamine: protection by 1-phenyl-3-(2-thiazolyl)-2-thiourea, diethyldithiocarbamate, methimazole, cysteamine, ethanol and n-butanol. J Pharmacol Exp Ther 199(2):336–352

  12. Cui Z-W, Xie Z-X, Wang B-F, Zhong Z-H, Chen X-Y et al (2015) Carvacrol protects neuroblastoma SH-SY5Y cells against Fe 2+-induced apoptosis by suppressing activation of MAPK/JNK-NF-κB signaling pathway. Acta Pharmacol Sin 36(12):1426

  13. Dati L, Ulrich H, Real C, Feng Z, Sun H, Britto LJN (2017) Carvacrol promotes neuroprotection in the mouse hemiparkinsonian model. Neuroscience 356:176–181

  14. Gasparotto J, Ribeiro CT, Bortolin RC, Somensi N, Rabelo TK et al (2017) Targeted inhibition of RAGE in substantia nigra of rats blocks 6-OHDA–induced dopaminergic denervation. Sci Rep 7(1):8795

  15. Gutteridge JM, Halliwell B (1990) The measurement and mechanism of lipid peroxidation in biological systems. Trends Biochem Sci 15(4):129–135

  16. Haddadi H, Rajaei Z, Alaei H, Shahidani S (2018) Chronic treatment with carvacrol improves passive avoidance memory in a rat model of Parkinson's disease. Arq Neuropsiquiatr 76(2):71–77. https://doi.org/10.1590/0004-282x20170193

  17. Haghdoost-Yazdi H, Piri H, Najafipour R, Faraji A, Fraidouni N, Dargahi T, Alipour Heidari M (2017) Blockade of fast A-type and TEA-sensitive potassium channels provide an antiparkinsonian effect in a 6-OHDA animal model. Neurosciences 22(1):44–50

  18. Hashemi M, Karami-Tehrani F, Ghavami S, Maddika S, Los M (2005) Adenosine and deoxyadenosine induces apoptosis in oestrogen receptor-positive and-negative human breast cancer cells via the intrinsic pathway. Cell Prolif 38(5):269–285

  19. Hayashi A, Matsunaga N, Okazaki H, Kakimoto K, Kimura Y, Azuma H, Ikeda E, Shiba T, Yamato M, Yamada KI, Koyanagi S, Ohdo S (2013) A disruption mechanism of the molecular clock in a MPTP mouse model of Parkinson’s disease. NeuroMolecular Med 15(2):238–251

  20. Held P, Newick K (2009) Using BioTek’s Synergy™ HT reader to measure reactive oxygen species (ROS) generation in stimulated cells. BioTechniques 46(1):61–62

  21. Hirsch EC, Vyas S, Hunot S (2012) Neuroinflammation in Parkinson’s disease. Parkinsonism Relat Disord 18:S210–S212

  22. Hornykiewicz O (1986) Biochemical pathophysiology of Parkinson’s disease. Adv Neurol 45:19–34

  23. Hosseini M, Rajaei Z, Alaei H, Tajadini M (2016) The effects of crocin on 6-OHDA-induced oxidative/nitrosative damage and motor behaviour in hemiparkinsonian rats. Malays J Med Sci 23(6):35–43

  24. Hotta M, Nakata R, Katsukawa M, Hori K, Takahashi S, Inoue H (2010) Carvacrol, a component of thyme oil, activates PPARα and γ and suppresses COX-2 expression. J Lipid Res 51(1):132–139. https://doi.org/10.1194/jlr.M900255-JLR200

  25. Iancu R, Mohapel P, Brundin P, Paul G (2005) Behavioral characterization of a unilateral 6-OHDA-lesion model of Parkinson’s disease in mice. Behav Brain Res 162(1):1–10

  26. Jalewa J, Sharma MK, Gengler S, Hölscher C (2017) A novel GLP-1/GIP dual receptor agonist protects from 6-OHDA lesion in a rat model of Parkinson’s disease. Neuropharmacology 117:238–248

  27. Janda E, Lascala A, Carresi C, Parafati M, Aprigliano S, Russo V, Savoia C, Ziviani E, Musolino V, Morani F, Isidoro C, Mollace V (2015) Parkinsonian toxin-induced oxidative stress inhibits basal autophagy in astrocytes via NQO2/quinone oxidoreductase 2: implications for neuroprotection. Autophagy 11(7):1063–1080

  28. Jayakumar S, Madankumar A, Asokkumar S, Raghunandhakumar S, Kamaraj S et al (2012) Potential preventive effect of carvacrol against diethylnitrosamine-induced hepatocellular carcinoma in rats. Mol Cell Biochem 360(1–2):51–60

  29. Jha SK, Jha NK, Kar R, Ambasta RK, Kumar P (2015) p38 MAPK and PI3K/AKT signalling cascades inParkinson’s disease. Int J Mol Cell Med 4(2):67–86

  30. Jiang ZS, Pu ZC, Hao ZH (2015) Carvacrol protects against spinal cord injury in rats via suppressing oxidative stress and the endothelial nitric oxide synthase pathway. Mol Med Rep 12(4):5349–5354

  31. Jollow D, Mitchell J, Zampaglione N a, Gillette J (1974) Bromobenzene-induced liver necrosis. Protective role of glutathione and evidence for 3, 4-bromobenzene oxide as the hepatotoxic metabolite. Pharmacology 11(3):151–169

  32. Kim M-K, Kim S-C, Kang J-I, Hyun J-H, Boo H-J, Eun SY, Park DB, Yoo ES, Kang HK, Kang JH (2011) 6-Hydroxydopamine-induced PC12 cell death is mediated by MEF2D down-regulation. Neurochem Res 36(2):223–231

  33. Lee J, Jung E, Yu H, Kim Y, Ha J, Kim YS, Park D (2008) Mechanisms of carvacrol-induced expression of type I collagen gene. J Dermatol Sci 52(3):160–169

  34. Lins L, Souza MF, Bispo JMM, Gois AM, Melo TCS et al (2018) Carvacrol prevents impairments in motor and neurochemical parameters in a model of progressive parkinsonism induced by reserpine. Brain Res Bull 139:9–15. https://doi.org/10.1016/j.brainresbull.2018.01.017

  35. Llana-Ruiz-Cabello M, Gutiérrez-Praena D, Pichardo S, Moreno FJ, Bermúdez JM, Aucejo S, Cameán AM (2014) Cytotoxicity and morphological effects induced by carvacrol and thymol on the human cell line Caco-2. Food Chem Toxicol 64:281–290

  36. Llana-Ruiz-Cabello M, Gutiérrez-Praena D, Puerto M, Pichardo S, Jos Á, Cameán AM (2015) In vitro pro-oxidant/antioxidant role of carvacrol, thymol and their mixture in the intestinal Caco-2 cell line. Toxicol in Vitro 29(4):647–656

  37. Magalingam KB, Radhakrishnan A, Haleagrahara N (2014) Protective effects of flavonol isoquercitrin, against 6-hydroxy dopamine (6-OHDA)-induced toxicity in PC12 cells. BMC Res Notes 7(1):49

  38. Melo FHC, Venâncio ET, De Sousa DP, Fonteles MMDF, De Vasconcelos SMM et al (2010) Anxiolytic-like effect of Carvacrol (5-isopropyl-2-methylphenol) in mice: involvement with GABAergic transmission. Fundam Clin Pharmacol 24(4):437–443

  39. Melo FHC, Moura BA, de Sousa DP, de Vasconcelos SMM, Macedo DS, Fonteles MMF, Viana GSB, de Sousa FCF (2011) Antidepressant-like effect of carvacrol (5-Isopropyl-2-methylphenol) in mice: involvement of dopaminergic system. Fundam Clin Pharmacol 25(3):362–367

  40. Mu X, He G, Cheng Y, Li X, Xu B, Du G (2009) Baicalein exerts neuroprotective effects in 6-hydroxydopamine-induced experimental parkinsonism in vivo and in vitro. Pharmacol Biochem Behav 92(4):642–648

  41. Neves KRT, Nobre HV, Leal LKA, de Andrade GM, Brito GADC, Viana GSDB (2015, 2015) Pentoxifylline neuroprotective effects are possibly related to its anti-inflammatory and TNF-alpha inhibitory properties, in the 6-OHDA model of Parkinson’s disease. Parkinsons Dis:108179

  42. Ogawa N, Hirose Y, Ohara S, Ono T, Watanabe Y (1985) A simple quantitative bradykinesia test in MPTP-treated mice. Res Commun Chem Pathol Pharmacol 50(3):435–441

  43. Olatunji OJ, Feng Y, Olatunji OO, Tang J, Ouyang Z, Su Z (2016) Cordycepin protects PC12 cells against 6-hydroxydopamine induced neurotoxicity via its antioxidant properties. Biomed Pharmacother 81:7–14

  44. Park G, Park Y-J, Yang HO, Oh MS (2013) Ropinirole protects against 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-induced neurotoxicity in mice via anti-apoptotic mechanism. Pharmacol Biochem Behav 104:163–168

  45. Paxinos G, Watson C (1998) A stereotaxic atlas of the rat brain. Academic, New York

  46. Przedbroski S, Leviver M, Jiang H, Ferreira M, Jackson-Lewis V, Donaldson D, Togasaki DM (1995) Dose-dependent lesions of the dopaminergic nigrostriatal pathway induced by intrastriatal injection of 6-hydroxydopamine. Neuroscience 67(3):631–647

  47. Sadegh M, Sakhaie MH (2018) Carvacrol mitigates proconvulsive effects of lipopolysaccharide, possibly through the hippocampal cyclooxygenase-2 inhibition. Metab Brain Dis 33(6):2045–2050

  48. Shah M, Rajagopalan S, Xu L, Voshavar C, Shurubor Y, Beal F, Andersen JK, Dutta AK (2014) The high-affinity D2/D3 agonist D512 protects PC12 cells from 6-OHDA-induced apoptotic cell death and rescues dopaminergic neurons in the MPTP mouse model of Parkinson’s disease. J Neurochem 131(1):74–85. https://doi.org/10.1111/jnc.12767

  49. Shimohama S, Sawada H, Kitamura Y, Taniguchi T (2003) Disease model: Parkinson’s disease. Trends Mol Med 9(8):360–365

  50. Singsai K, Akaravichien T, Kukongviriyapan V, Sattayasai J (2015) Protective effects of Streblus asper leaf extract on H2O2-induced ROS in SK-N-SH cells and MPTP-induced Parkinson’s disease-like symptoms in C57BL/6 mouse. Evid Based Complement Alternat Med 2015:970354

  51. Slater T (1984) Overview of methods used for detecting lipid peroxidation. In: Methods in enzymology, vol 105. Elsevier, Amsterdam, pp 283–293

  52. Sofic E, Lange KW, Jellinger K, Riederer P (1992) Reduced and oxidized glutathione in the substantia nigra of patients with Parkinson’s disease. Neurosci Lett 142(2):128–130

  53. Soto-Otero R, Méndez-Álvarez E, Hermida-Ameijeiras Á, Muñoz-Patiño AM, Labandeira-Garcia JL (2000) Autoxidation and neurotoxicity of 6-hydroxydopamine in the presence of some antioxidants. J Neurochem 74(4):1605–1612. https://doi.org/10.1046/j.1471-4159.2000.0741605.x

  54. Tao L, Li X, Zhang L, Tian J, Li X, Sun X, Li X, Jiang L, Zhang X, Chen J (2011) Protective effect of tetrahydroxystilbene glucoside on 6-OHDA-induced apoptosis in PC12 cells through the ROS-NO pathway. PLoS One 6(10):e26055

  55. Türkez H, Aydın E (2016) Investigation of cytotoxic, genotoxic and oxidative properties of carvacrol in human blood cells. Toxicol Ind Health 32(4):625–33

  56. Ultee A, Kets E, Smid E (1999) Mechanisms of action of carvacrol on the food-borne pathogen Bacillus cereus. Appl Environ Microbiol 65(10):4606–4610

  57. Viveros-Paredes JM, González-Castañeda RE, Gertsch J, Chaparro-Huerta V, López-Roa RI et al (2017) Neuroprotective effects of β-caryophyllene against dopaminergic neuron injury in a murine model of Parkinson’s disease induced by MPTP. Pharmaceuticals 10(3):60

  58. Wang P, Luo Q, Qiao H, Ding H, Cao Y et al (2017, 2017) The neuroprotective effects of carvacrol on ethanol-induced hippocampal neurons impairment via the antioxidative and antiapoptotic pathways. Oxidative Med Cell Longev:4079425

  59. Ximenes JCM, Neves KRT, Leal LKA, do Carmo MRS, Brito GADC et al (2015) Valproic acid neuroprotection in the 6-OHDA model of Parkinson’s disease is possibly related to its anti-inflammatory and HDAC inhibitory properties. J Neurodegener Dis 2015:313702

  60. Yu H, Zhang Z-L, Chen J, Pei A, Hua F, Qian X, He J, Liu CF, Xu X (2012) Carvacrol, a food-additive, provides neuroprotection on focal cerebral ischemia/reperfusion injury in mice. PLoS One 7(3):e33584

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Correspondence to Anahita Torkaman-Boutorabi.

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The experimental procedures were carried out in accordance with international guideline for the care and use of laboratory animals, and all efforts were made to minimize animal suffering and reduce the number of animals used in the experiments. The Research and Ethics Committee of Tehran University of Medical Sciences, School of Advanced Technologies in Medicine, approved the experimental protocol.

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Manouchehrabadi, M., Farhadi, M., Azizi, Z. et al. Carvacrol Protects Against 6-Hydroxydopamine-Induced Neurotoxicity in In Vivo and In Vitro Models of Parkinson’s Disease. Neurotox Res 37, 156–170 (2020). https://doi.org/10.1007/s12640-019-00088-w

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Keywords

  • CAR
  • Neuroprotection
  • 6-OHDA
  • Parkinson’s disease
  • PC12
  • Rat