Carvacrol Depends on Heme Oxygenase-1 (HO-1) to Exert Antioxidant, Anti-inflammatory, and Mitochondria-Related Protection in the Human Neuroblastoma SH-SY5Y Cells Line Exposed to Hydrogen Peroxide
The link between mitochondrial dysfunction, redox impairment, and inflammation leads to increased rates of brain cells loss in neurodegenerative diseases and in affective disorders. Carvacrol (CAR) is a component of essential oils found in Labiatae. CAR exerts antioxidant and anti-inflammatory effects in different cell types, as assessed in both in vitro and in vivo experimental designs. Nonetheless, it was not previously investigated whether and how CAR would prevent mitochondrial impairment in human cells exposed to a pro-oxidant challenge. Therefore, we analyzed here whether a pretreatment (for 4 h) with CAR (10–1000 µM) would promote mitochondrial protection in the human neuroblastoma cells SH-SY5Y exposed to hydrogen peroxide (H2O2). We found that CAR at 100 µM prevented the H2O2-induced decline in the activity of the complexes I and V, as well as on the levels of adenosine triphosphate (ATP). CAR also prevented the H2O2-elicited decrease in the activity of the mitochondrial enzymes aconitase, α-ketoglutarate dehydrogenase, and succinate dehydrogenase. Moreover, CAR induced an antioxidant action by decreasing the levels of lipid peroxidation, protein carbonylation, and protein nitration in the mitochondrial membranes. Interestingly, CAR prevented the pro-inflammatory action of H2O2 by downregulating the transcription factor nuclear factor-κB (NF-κB). The inhibition of the heme oxygenase-1 (HO-1) enzyme by zinc protoporphyrin IX (ZnPP IX, 10 µM) suppressed the preventive effects caused by CAR regarding mitochondrial function and inflammation. Thus, it is suggested that CAR caused cytoprotective effects by an HO-1-dependent manner in SH-SY5Y cells exposed to H2O2.
KeywordsCarvacrol Mitochondria Bioenergetics Antioxidant Anti-inflammatory HO-1
This work was supported by the Conselho Nacional de Pesquisa e Desenvolvimento Tecnológico (CNPq; Edital Universal 2016; 400216/2016-7) and by the Fundação de Apoio à Pesquisa do Estado de Mato Grosso (FAPEMAT; Edital Universal; 222541/2015). ALC receives a CAPES fellow (Bolsa de Mestrado).
Compliance with Ethical Standards
Conflict of interest
All authors declare that they have no conflict of interest.
- 15.Sies H, Berndt C, Jones DP (2017) Oxidative stress. Annu Rev Biochem 86:715–748. https://doi.org/10.1146/annurev-biochem-061516-045037 CrossRefGoogle Scholar
- 16.de Oliveira MR, Ferreira GC, Schuck PF, Dal Bosco SM (2015) Role for the PI3K/Akt/Nrf2 signaling pathway in the protective effects of carnosic acid against methylglyoxal-induced neurotoxicity in SH-SY5Y neuroblastoma cells. Chem Biol Interact 242:396–406. https://doi.org/10.1016/j.cbi.2015.11.003 CrossRefGoogle Scholar
- 18.de Oliveira MR, Fürstenau CR, de Souza ICC, da Costa Ferreira G (2017) Tanshinone I attenuates the effects of a challenge with H2O2 on the functions of tricarboxylic acid cycle and respiratory chain in SH-SY5Y cells. Mol Neurobiol 54:7858–7868. https://doi.org/10.1007/s12035-016-0267-7 CrossRefGoogle Scholar
- 22.de Oliveira MR, Peres A, Ferreira GC, Schuck PF, Gama CS, Bosco SMD (2017) Carnosic acid protects mitochondria of human neuroblastoma SH-SY5Y cells exposed to paraquat through activation of the Nrf2/HO-1 Axis. Mol Neurobiol 54:5961–5972. https://doi.org/10.1007/s12035-016-0100-3 CrossRefGoogle Scholar
- 32.Cui ZW, Xie ZX, Wang BF, Zhong ZH, Chen XY, Sun YH, Sun QF, Yang GY, Bian LG (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 36:1426–1436. https://doi.org/10.1038/aps.2015.90 CrossRefPubMedCentralGoogle Scholar
- 40.de Oliveira MR, da Rocha RF, Pasquali MA, Moreira JC (2012) The effects of vitamin A supplementation for 3 months on adult rat nigrostriatal axis: increased monoamine oxidase enzyme activity, mitochondrial redox dysfunction, increased β-amyloid(1–40) peptide and TNF-α contents, and susceptibility of mitochondria to an in vitro H2O2 challenge. Brain Res Bull 87:432–444. https://doi.org/10.1016/j.brainresbull.2012.01.005 CrossRefGoogle Scholar
- 48.Qin W, Zhang J, Lv W, Wang X, Sun B (2013) Effect of carbon monoxide-releasing molecules II-liberated CO on suppressing inflammatory response in sepsis by interfering with nuclear factor kappa B activation. PLoS One 8:e75840. https://doi.org/10.1371/journal.pone.0075840 CrossRefPubMedCentralGoogle Scholar
- 53.Niture SK, Khatri R, Jaiswal AK (2014) Regulation of Nrf2-an update. Free Radic Biol Med 66:36–44. https://doi.org/10.1016/j.freeradbiomed.2013.02.008 CrossRefGoogle Scholar
- 58.Steiner J, Walter M, Gos T, Guillemin GJ, Bernstein HG, Sarnyai Z, Mawrin C, Brisch R, Bielau H, Meyer zu Schwabedissen L, Bogerts B, Myint AM (2011) Severe depression is associated with increased microglial quinolinic acid in subregions of the anterior cingulate gyrus: evidence for an immune-modulated glutamatergic neurotransmission? J Neuroinflammation 8:94. https://doi.org/10.1186/1742-2094-8-94 CrossRefPubMedCentralGoogle Scholar