Top

Published in:

01-03-2022 | Original Article

A novel antagonist of TRPM2 and TRPV4 channels: Carvacrol

Author: Mustafa Nazıroğlu

Published in: Metabolic Brain Disease | Issue 3/2022

Abstract

The overload cytosolic free Ca²⁺ (cCa²⁺) influx-mediated excessive generation of oxidative stress in the pathophysiological conditions induces neuronal and cellular injury via the activation of cation channels. TRPM2 and TRPV4 channels are activated by oxidative stress, and their specific antagonists have not been discovered yet. The antioxidant and anti-Covid-19 properties of carvacrol (CARV) were recently reported. Hence, I suspected possible antagonist properties of CARV against oxidative stress (OS)/ADP-ribose (ADPR)-induced TRPM2 and GSK1016790A (GSK)-mediated TRPV4 activations in neuronal and kidney cells. I investigated the antagonist role of CARV on the activations of TRPM2 and TRPV4 in SH-SY5Y neuronal, BV-2 microglial, and HEK293 cells. The OS/ADPR and GSK in the cells caused to increase of TRPM2/TRPV4 current densities and overload cytosolic free Ca²⁺ (cCa²⁺) influx with an increase of mitochondrial membrane potential, cytosolic (cROS), and mitochondrial (mROS) ROS. The changes were not observed in the absence of TRPM2 and TRPV4 or the presence of Ca²⁺ free extracellular buffer and PARP-1 inhibitors (PJ34 and DPQ). When OS-induced TRPM2 and GSK-induced TRPV4 activations were inhibited by the treatment of CARV, the increase of cROS, mROS, lipid peroxidation, apoptosis, cell death, cCa²⁺ concentration, caspase -3, and caspase -9 levels were restored via upregulation of glutathione and glutathione peroxidase. In conclusion, the treatment of CARV modulated the TRPM2 and TRPV4-mediated overload Ca²⁺ influx and may provide an avenue for protecting TRPM2 and TRPV4-mediated neurodegenerative diseases associated with the increase of mROS and cCa²⁺.

Graphical abstract

The possible TRPM2 and TRPV4 blocker action of carvacrol (CARV) via the modulation oxidative stress and apoptosis in the SH-SY5Y neuronal cells. TRPM2 is activated by DNA damage-induced (via PARP-1 activation) ADP-ribose (ADPR) and reactive oxygen species (ROS) (H₂O₂), although it is inhibited by nonspecific inhibitors (ACA and 2-APB). TRPV4 is activated by the treatments of GSK1016790A (GSK), although it is inhibited by a nonspecific inhibitor (ruthenium red, RuRe). The treatment of GSK induces excessive generation of ROS. The accumulation of free cytosolic Ca²⁺ (cCa²⁺) via the activations of TRPM2 and TRPV4 in the mitochondria causes the increase of mitochondrial membrane depolarization (ΔΨm). In turn, the increase of ΔΨm causes the excessive generation of ROS. The TRPM2 and TRPV4-induced the excessive generations of ROS result in the increase of apoptosis and cell death via the activations of caspase -3 (Casp-3) and caspase -9 (Casp-9) in the neuronal cells, although their oxidant actions decrease the glutathione (GSH) and glutathione peroxidase (GSHPx) levels. The oxidant and apoptotic adverse actions of TRPM2 and TRPV4 are modulated by the treatment of CARV

Available only for authorised users

Akpınar H, Nazıroğlu M, Övey İS, Çiğ B, Akpınar O (2016) The neuroprotective action of dexmedetomidine on apoptosis, calcium entry and oxidative stress in cerebral ischemia-induced rats: Contribution of TRPM2 and TRPV1 channels. Sci Rep 6:37196. https://doi.org/10.1038/srep37196CrossRefPubMedPubMedCentral

Akyuva Y, Nazıroğlu M (2020) Resveratrol attenuates hypoxia-induced neuronal cell death, inflammation and mitochondrial oxidative stress by modulation of TRPM2 channel. Sci Rep 10(1):6449. https://doi.org/10.1038/s41598-020-63577-5CrossRefPubMedPubMedCentral

Armağan HH, Nazıroğlu M (2021) Curcumin attenuates hypoxia-Induced oxidative neurotoxicity, apoptosis, calcium, and zinc ion influxes in a neuronal cell line: Involvement of TRPM2 channel. Neurotox Res 39(3):618–633. https://doi.org/10.1007/s12640-020-00314-wCrossRefPubMed

Arunasree KM (2010) Anti-proliferative effects of carvacrol on a human metastatic breast cancer cell line, MDA-MB 231. Phytomedicine 17(8–9):581–588. https://doi.org/10.1016/j.phymed.2009.12.008CrossRefPubMed

Badr H, Kozai D, Sakaguchi R, Numata T, Mori Y (2016) Different Contribution of Redox-Sensitive Transient Receptor Potential Channels to Acetaminophen-Induced Death of Human Hepatoma Cell Line. Front Pharmacol 7:19. https://doi.org/10.3389/fphar.2016.00019CrossRefPubMedPubMedCentral

Banik S, Akter M, Corpus Bondad SE, Saito T, Hosokawa T, Kurasaki M (2019) Carvacrol inhibits cadmium toxicity through combating against caspase dependent/independent apoptosis in PC12 cells. Food Chem Toxicol 134:110835. https://doi.org/10.1016/j.fct.2019.110835CrossRef

Bao L, Chen SJ, Conrad K et al (2016) Depletion of the human ion channel TRPM2 in neuroblastoma demonstrates its key role in cell survival through modulation of mitochondrial reactive oxygen species and bioenergetics. J Biol Chem 291(47):24449–24464. https://doi.org/10.1074/jbc.M116.747147CrossRefPubMedPubMedCentral

Bollimuntha S, Singh BB, Shavali S, Sharma SK, Ebadi M (2005) TRPC1-mediated inhibition of 1-methyl-4-phenylpyridinium ion neurotoxicity in human SH-SY5Y neuroblastoma cells. J Biol Chem 280(3):2132–2140. https://doi.org/10.1074/jbc.M407384200CrossRefPubMed

Bubolz AH, Mendoza SA, Zheng X, Zinkevich NS, Li R, Gutterman DD, Zhang DX (2012) Activation of endothelial TRPV4 channels mediates flow-induced dilation in human coronary arterioles: role of Ca2+ entry and mitochondrial ROS signaling. Am J Physiol Heart Circ Physiol 302(3):H634–H642. https://doi.org/10.1152/ajpheart.00717.2011CrossRefPubMed

Butenko O, Dzamba D, Benesova J, Honsa P, Benfenati V, Rusnakova V et al (2012) The increased activity of TRPV4 channel in the astrocytes of the adult rat hippocampus after cerebral hypoxia/ischemia. PLoS One 7(6):e39959. https://doi.org/10.1371/journal.pone.0039959CrossRefPubMedPubMedCentral

Carrasco C, Nazıroǧlu M, Rodríguez AB, Pariente JA (2018) Neuropathic Pain: Delving into the Oxidative Origin and the Possible Implication of Transient Receptor Potential Channels. Front Physiol 9:95. https://doi.org/10.3389/fphys.2018.00095CrossRefPubMedPubMedCentral

Chen W, Xu B, Xiao A, Liu L, Fang X, Liu R et al (2015) TRPM7 inhibitor carvacrol protects brain from neonatal hypoxic-ischemic injury. Mol Brain 8:11. https://doi.org/10.1186/s13041-015-0102-5CrossRefPubMedPubMedCentral

Chen Y, Ba L, Huang W, Liu Y, Pan H, Mingyao E et al (2017) Role of carvacrol in cardioprotection against myocardial ischemia/reperfusion injury in rats through activation of MAPK/ERK and Akt/eNOS signaling pathways. Eur J Pharmacol 796:90–100. https://doi.org/10.1016/j.ejphar.2016.11.053CrossRefPubMed

de la Monte SM, Neely TR, Cannon J, Wands JR (2000) Oxidative stress and hypoxia-like injury cause Alzheimer-type molecular abnormalities in central nervous system neurons. Cell Mol Life Sci 57(10):1471–1481. https://doi.org/10.1007/PL00000630CrossRefPubMed

Dong Q, Li J, Wu QF, Zhao N, Qian C, Ding D et al (2017) Blockage of transient receptor potential vanilloid 4 alleviates myocardial ischemia/reperfusion injury in mice. Sci Rep 7:42678. https://doi.org/10.1038/srep42678CrossRefPubMedPubMedCentral

Donkó A, Ruisanchez E, Orient A, Enyedi B, Kapui R, Péterfi Z et al (2010) Urothelial cells produce hydrogen peroxide through the activation of Duox1. Free Radic Biol Med 49(12):2040–2048. https://doi.org/10.1016/j.freeradbiomed.2010.09.027CrossRefPubMed

Dringen R (2000) Metabolism and functions of glutathione in brain. Prog Neurobiol 62(6):649–671. https://doi.org/10.1016/S0301-0082(99)00060-XCrossRefPubMed

Du J, Wong WY, Sun L, Huang Y, Yao X (2012) Protein kinase G inhibits flow-induced Ca2+ entry into collecting duct cells. J Am Soc Nephrol 23(7):1172–1180. https://doi.org/10.1681/ASN.2011100972CrossRefPubMedPubMedCentral

Ertilav K (2019) Pregabalin protected cisplatin-induced oxidative neurotoxicity in neuronal cell line. J Cell Neurosci Oxid Stress 11(1):815–824. https://doi.org/10.37212/jcnos.653500CrossRef

Espino J, Bejarano I, Paredes SD, González D, Barriga C, Reiter RJ et al (2010) Melatonin counteracts alterations in oxidative metabolism and cell viability induced by intracellular calcium overload in human leucocytes: changes with age. Basic Clin Pharmacol Toxicol 107(1):590–597. https://doi.org/10.1111/j.1742-7843.2010.00546.xCrossRefPubMed

Fonfria E, Marshall IC, Benham CD, Boyfield I, Brown JD, Hill K et al (2004) TRPM2 channel opening in response to oxidative stress is dependent on activation of poly(ADP-ribose) polymerase. Br J Pharmacol 143(1):186–192CrossRef

Guan X, Li X, Yang X, Yan J, Shi P, Ba L, Cao Y, Wang P (2019) The neuroprotective effects of carvacrol on ischemia/reperfusion-induced hippocampal neuronal impairment by ferroptosis mitigation. Life Sci 235:116795. https://doi.org/10.1016/j.lfs.2019.116795CrossRefPubMed

Guse AH (2015) Calcium mobilizing second messengers derived from NAD. Biochim Biophys Acta 1854(9):1132–1137. https://doi.org/10.1016/j.bbapap.2014.12.015CrossRefPubMed

Halliwell B (2006) Oxidative stress and neurodegeneration: where are we now? J Neurochem 97(6):1634–1658. https://doi.org/10.1111/j.1471-4159.2006.03907.xCrossRefPubMed

Hara Y, Wakamori M, Ishii M, Maeno E, Nishida M, Yoshida T et al (2002) LTRPC2 Ca2+-permeable channel activated by changes in redox status confers susceptibility to cell death. Mol Cell 9(1):163–173. https://doi.org/10.1016/S1097-2765(01)00438-5CrossRefPubMed

Hong Z, Tian Y, Yuan Y, Qi M, Li Y, Du Y et al (2016) Enhanced oxidative stress is responsible for TRPV4-induced neurotoxicity. Front Cell Neurosci 10:232. https://doi.org/10.3389/fncel.2016.00232CrossRefPubMedPubMedCentral

Islam MT (2017) Oxidative stress and mitochondrial dysfunction-linked neurodegenerative disorders. Neurol Res 39(1):73–82. https://doi.org/10.1080/01616412.2016.1251711CrossRefPubMed

Javed H, Meeran MFN, Jha NK, Ojha S (2021) Carvacrol, a plant metabolite targeting viral protease (M^pro) and ACE2 in host cells can be a possible candidate for COVID-19. Front Plant Sci 11:601335. https://doi.org/10.3389/fpls.2020.601335CrossRefPubMedPubMedCentral

Jeong JH, Lee SH, Kho AR, Hong DK, Kang DH, Kang BS et al (2020) The Transient Receptor Potential Melastatin 7 (TRPM7) inhibitors suppress seizure-induced neuron death by inhibiting zinc neurotoxicity. Int J Mol Sci 21(21):7897. https://doi.org/10.3390/ijms21217897CrossRefPubMedCentral

Ji SG, Medvedeva YV, Weiss JH (2020) Zn²⁺ entry through the mitochondrial calcium uniporter is a critical contributor to mitochondrial dysfunction and neurodegeneration. Exp Neurol 325:113161. https://doi.org/10.1016/j.expneurol.2019.113161CrossRefPubMed

Jie P, Hong Z, Tian Y, Li Y, Lin L et al (2015) Activation of transient receptor potential vanilloid 4 induces apoptosis in hippocampus through downregulating PI3K/Akt and upregulating p38 MAPK signaling pathways. Cell Death Dis 6(6):e1775. https://doi.org/10.1038/cddis.2015.146CrossRefPubMedPubMedCentral

Jie P, Lu Z, Hong Z, Li L, Zhou L, Li Y et al (2016) Activation of transient receptor potential vanilloid 4 is involved in neuronal injury in middle cerebral artery occlusion in mice. Mol Neurobiol 53(1):8–17. https://doi.org/10.1007/s12035-014-8992-2CrossRefPubMed

Kahya MC, Nazıroğlu M, Övey İS (2017) Modulation of diabetes-induced oxidative Stress, apoptosis, and Ca(2+) entry through TRPM2 and TRPV1 channels in dorsal root ganglion and hippocampus of diabetic rats by melatonin and selenium. Mol Neurobiol 54(3):2345–2360. https://doi.org/10.1007/s12035-016-9727-3CrossRefPubMed

Kumamoto E, Fujita T (2016) Differential activation of TRP channels in the adult rat spinal substantia gelatinosa by stereoisomers of plant-derived chemicals. Pharmaceuticals (Basel) 9(3):46. https://doi.org/10.3390/ph9030046CrossRef

Lipski J, Park TI, Li D, Lee SC, Trevarton AJ, Chung KK et al (2006) Involvement of TRP-like channels in the acute ischemic response of hippocampal CA1 neurons in brain slices. Brain Res 1077(1):187–199. https://doi.org/10.1016/j.brainres.2006.01.016CrossRefPubMed

Malko P, Ding R, Jiang LH (2021) TRPM2 channel in oxidative stress-induced mitochondrial dysfunction and apoptotic cell death. Adv Protein Chem Struct Biol 125:51–72. https://doi.org/10.1016/bs.apcsb.2020.12.001CrossRefPubMed

McHugh D, Flemming R, Xu SZ, Perraud AL, Beech DJ (2003) Critical intracellular Ca2+ dependence of transient receptor potential melastatin 2 (TRPM2) cation channel activation. J Biol Chem 278(13):11002–11006. https://doi.org/10.1074/jbc.M210810200CrossRefPubMed

Naeem K, Tariq Al Kury L, Nasar F, Alattar A, Alshaman R et al (2021) Natural dietary supplement, carvacrol, alleviates LPS-induced oxidative stress, neurodegeneration, and depressive-like behaviors via the Nrf2/HO-1 pathway. J Inflamm Res 14:1313–1132. https://doi.org/10.2147/JIR.S294413CrossRefPubMedPubMedCentral

Nakagawa F, Higashi S, Ando E, Ohsumi T, Watanabe S, Takeuchi H (2020) Modification of TRPV4 activity by acetaminophen. Heliyon 6(1):e03301. https://doi.org/10.1016/j.heliyon.2020.e03301CrossRefPubMedPubMedCentral

Nazıroğlu M, Dikici DM, Dursun S (2012) Role of oxidative stress and Ca²⁺ signaling on molecular pathways of neuropathic pain in diabetes: focus on TRP channels. Neurochem Res 37(10):2065–2075. https://doi.org/10.1007/s11064-012-0850-xCrossRefPubMed

Nazıroğlu M, Lückhoff A (2008) Effects of antioxidants on calcium influx through TRPM2 channels in transfected cells activated by hydrogen peroxide. J Neurol Sci 270(1–2):152–158. https://doi.org/10.1016/j.jns.2008.03.003CrossRefPubMed

Nazıroğlu M, Öz A, Yıldızhan K (2020) Selenium and neurological diseases: Focus on peripheral pain and TRP channels. Curr Neuropharmacol 18(6):501–517. https://doi.org/10.2174/1570159X18666200106152631CrossRefPubMedPubMedCentral

Nazıroğlu M (2007) New molecular mechanisms on the activation of TRPM2 channels by oxidative stress and ADP-ribose. Neurochem Res 32(11):1990–2001CrossRef

Özkaya D, Shu X, Nazıroğlu M (2021) Deletion of mitochondrial translocator protein (TSPO) gene decreases oxidative retinal pigment epithelial cell death via modulation of TRPM2 channel. Biology (Basel) 10(5):382. https://doi.org/10.3390/biology10050382CrossRef

Pacheco G, Oliveira AP, Noleto IRSG, Araújo AK, Lopes ALF, Sousa FBM et al (2021) Activation of transient receptor potential vanilloid channel 4 contributes to the development of ethanol-induced gastric injury in mice. Eur J Pharmacol 902:174113. https://doi.org/10.1016/j.ejphar.2021.174113CrossRefPubMed

Parnas M, Peters M, Dadon D, Lev S, Vertkin I, Slutsky I, Minke B (2009) Carvacrol is a novel inhibitor of Drosophila TRPL and mammalian TRPM7 channels. Cell Calcium 45(3):300–309. https://doi.org/10.1016/j.ceca.2008.11.009CrossRefPubMedPubMedCentral

Pathania AS, Guru SK, Verma MK, Sharma C, Abdullah ST, Malik F et al (2013) Disruption of the PI3K/AKT/mTOR signaling cascade and induction of apoptosis in HL-60 cells by an essential oil from Monarda citriodora. Food Chem Toxicol 62:246–254. https://doi.org/10.1016/j.fct.2013.08.037CrossRefPubMed

Perraud AL, Fleig A, Dunn CA, Bagley LA, Launay P, Schmitz C et al (2001) ADP-ribose gating of the calcium-permeable LTRPC2 channel revealed by Nudix motif homology. Nature 411(6837):595–599. https://doi.org/10.1038/35079100CrossRefPubMed

Saleh J, Peyssonnaux C, Singh KK, Edeas M (2020) Mitochondria and microbiota dysfunction in COVID-19 pathogenesis. Mitochondrion 54:1–7. https://doi.org/10.1016/j.mito.2020.06.008CrossRefPubMedPubMedCentral

Sánchez JC, Muñoz LV, Ehrlich BE (2020) Modulating TRPV4 channels with paclitaxel and lithium. Cell Calcium 91:102266. https://doi.org/10.1016/j.ceca.2020.102266CrossRefPubMed

Schenk SA, Dick F, Herzog C, Eberhardt MJ, Leffler A (2019) Active metabolites of dipyrone induce a redox-dependent activation of the ion channels TRPA1 and TRPV1. Pain Rep 4(3):e720. https://doi.org/10.1097/PR9.0000000000000720CrossRefPubMedPubMedCentral

Sensi SL, Ton-That D, Sullivan PG, Jonas EA, Gee KR, Kaczmarek LK, Weiss JH (2003) Modulation of mitochondrial function by endogenous Zn2+ pools. Proc Natl Acad Sci U S A 100(10):6157–6162. https://doi.org/10.1073/pnas.1031598100CrossRefPubMedPubMedCentral

Sha’fie MSA, Rathakrishnan S, Hazanol IN, Dali MHI, Khayat ME, Ahmad S et al (2020) Ethanol induces microglial cell death via the NOX/ROS/PARP/TRPM2 signalling pathway. Antioxidants (Basel) 9(12):1253. https://doi.org/10.3390/antiox9121253CrossRef

Shahidullah M, Mandal A, Delamere NA (2012) TRPV4 in porcine lens epithelium regulates hemichannel-mediated ATP release and Na-K-ATPase activity. Am J Physiol Cell Physiol 302(12):C1751–C1761. https://doi.org/10.1152/ajpcell.00010.2012CrossRefPubMedPubMedCentral

Shi J, Mori E, Mori Y, Mori M, Li J, Ito Y, Inoue R (2004) Multiple regulation by calcium of murine homologues of transient receptor potential proteins TRPC6 and TRPC7 expressed in HEK293 cells. J Physiol 561(Pt 2):415–432. https://doi.org/10.1113/jphysiol.2004.075051CrossRefPubMedPubMedCentral

Skrzypski M, Kakkassery M, Mergler S, Grötzinger C, Khajavi N, Sassek M et al (2003) Activation of TRPV4 channel in pancreatic INS-1E beta cells enhances glucose-stimulated insulin secretion via calcium-dependent mechanisms. FEBS Lett 587(19):3281–3287. https://doi.org/10.1016/j.febslet.2013.08.025CrossRef

Takata T, Araki S, Tsuchiya Y, Watanabe Y (2020) Oxidative stress orchestrates MAPK and nitric-oxide synthase signal. Int J Mol Sci 21(22):8750. https://doi.org/10.3390/ijms21228750CrossRefPubMedCentral

Tian Y, Qi M, Hong Z, Li Y, Yuan Y, Du Y, Chen L, Chen L (2017) Activation of transient receptor potential vanilloid 4 promotes the proliferation of stem cells in the adult hippocampal dentate gyrus. Mol Neurobiol 54(8):5768–5779. https://doi.org/10.1007/s12035-016-0113-yCrossRefPubMed

Wang Z, Zhou L, An D, Xu W, Wu C, Sha S et al (2019) TRPV4-induced inflammatory response is involved in neuronal death in pilocarpine model of temporal lobe epilepsy in mice. Cell Death Dis 10(6):386. https://doi.org/10.1038/s41419-019-1612-3CrossRefPubMedPubMedCentral

Watanabe H, Davis JB, Smart D, Jerman JC, Smith GD, Hayes P et al (2002) Activation of TRPV4 channels (hVRL-2/mTRP12) by phorbol derivatives. J Biol Chem 277(16):13569–13577. https://doi.org/10.1074/jbc.M200062200CrossRefPubMed

Yıldızhan K, Nazıroğlu M (2020) Glutathione depletion and parkinsonian neurotoxin MPP⁺-induced TRPM2 channel activation play central roles in oxidative cytotoxicity and inflammation in microglia. Mol Neurobiol 57(8):3508–3525. https://doi.org/10.1007/s12035-020-01974-7CrossRefPubMed

Title: A novel antagonist of TRPM2 and TRPV4 channels: Carvacrol
Author: Mustafa Nazıroğlu
Publication date: 01-03-2022
Publisher: Springer US
Published in: Metabolic Brain Disease / Issue 3/2022
Print ISSN: 0885-7490
Electronic ISSN: 1573-7365
DOI: https://doi.org/10.1007/s11011-021-00887-1

At a glance: The STEP trials

Springer Medicine

A novel antagonist of TRPM2 and TRPV4 channels: Carvacrol

Abstract

Graphical abstract

At a glance: The STEP trials

Springer Medicine

Abstract

Graphical abstract

Please log in to get access to this content

Other articles of this Issue 3/2022

LncRNA NEAT1 ameliorate ischemic stroke via promoting Mfn2 expression through binding to Nova and activates Sirt3

Knockdown of circ_0007290 alleviates oxygen-glucose deprivation-induced neuronal injury by regulating miR-496/PDCD4 axis

The association of stem cell factor and soluble c-Kit (s-cKit) receptor serum concentrations with the severity and risk prediction of autism spectrum disorders

Correction to: RNA as a Source of Biomarkers for Amyotrophic Lateral Sclerosis

Sesamol prevents mitochondrial impairment and pro-inflammatory alterations in the human neuroblastoma SH-SY5Y cells: role for Nrf2

A kinetic scheme to examine the role of glial cells in the pathogenesis of Alzheimer’s disease