Skip to main content
SpringerLink
Log in

Thymoquinone reduces mitochondrial damage and death of cardiomyocytes induced by clozapine

  • Original Article
  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Abstract

The generation of a reactive nitrenium ion by microsomal/mitochondrial cytochrome P450 (CYPs) from clozapine (CLZ) has been suggested as the main cause of cardiotoxicity by this drug. Previous studies indicated that thymoquinone (TQ) as an active constituent of Nigella sativa has pharmacological effects such as antioxidant, reactive oxygen species (ROS) scavenger, and inhibitory effect on CYPs enzymes. Therefore, we hypothesized that TQ with these pharmacological effects can reduce CLZ-induced toxicity in isolated cardiomyocytes and mitochondria. Rat left ventricular cardiomyocytes and mitochondria were isolated by collagenase perfusion and differential centrifugation respectively. Then, isolated cardiomyocytes and mitochondria were pretreated with different concentrations of TQ (1, 5, and 10 μmol/l) for 30 min and then followed by exposure to CLZ (50 μmol/l) for 6 h. After 6 h of incubation, using biochemical evaluations and flow cytometric analysis, the parameters of cellular toxicity including cytotoxicity, the level of oxidized/reduced glutathione (GSH/GSSG), malondialdehyde (MDA), reactive oxygen species (ROS) formation, lysosomal membrane integrity, mitochondria membrane potential (ΔΨm) collapse, and mitochondrial toxicity including succinate dehydrogenase (SDH) activity and mitochondrial swelling were analyzed. We observed a significant toxicity in isolated cardiomyocytes and mitochondria after exposure with CLZ which was related to ROS formation, oxidative stress, GSH depletion, lysosomal and mitochondrial damages, and mitochondrial dysfunction and swelling, while TQ pretreatment reverted the above toxic effect of CLZ on isolated cardiomyocytes and mitochondria. Our results indicate that TQ prevents and reverses CLZ-induced cytotoxicity and mitochondrial damages in isolated cardiomyocytes and mitochondria, providing an experimental basis for clinical treatment on CLZ-induced cardiotoxicity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Data availability

Not applicable.

References

  • Abdel Hamid OI, Ahmed MG, Hassaneine HM, Rashed HE (2017) Evaluation of the role of captopril on clozapine-induced cardiotoxicity and hematotoxicity in adult male albino rats. Toxicol Res Appl 1:2397847317696539

    Google Scholar 

  • Ahmad S, Beg ZH (2013) Elucidation of mechanisms of actions of thymoquinone-enriched methanolic and volatile oil extracts from Nigella sativa against cardiovascular risk parameters in experimental hyperlipidemia. Lipids Health Dis 12:86

    Article  Google Scholar 

  • Albassam AA, Ahad A, Alsultan A, Al-Jenoobi FI (2018) Inhibition of cytochrome P450 enzymes by thymoquinone in human liver microsomes. Saudi Pharm J 26:673–677

    Article  Google Scholar 

  • Alberich S, Fernández-Sevillano J, González-Ortega I, Usall J, Sáenz M, González-Fraile E, González-Pinto A (2019) A systematic review of sex-based differences in effectiveness and adverse effects of clozapine. Psychiatry Res 280:112506

    Article  Google Scholar 

  • Arzuk E, Karakuş F, Orhan H (2021) Bioactivation of clozapine by mitochondria of the murine heart: possible cause of cardiotoxicity. Toxicology 447:152628

  • Atkin K, Kendall F, Gould D, Freeman H, Lieberman J, O’Sullivan D (1996) Neutropenia and agranulocytosis in patients receiving clozapine in the UK and Ireland. Br J Psychiatry 169:483–488

    Article  CAS  Google Scholar 

  • Badary OA, Taha RA, Gamal El-Din AM, Abdel-Wahab MH (2003) Thymoquinone is a potent superoxide anion scavenger. Drug Chem Toxicol 26:87–98

    Article  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • De Berardis D et al (2012) Update on the adverse effects of clozapine: focus on myocarditis. Curr Drug Safety 7:55–62

    Article  Google Scholar 

  • Dragovic S, Boerma JS, van Bergen L, Vermeulen NP, Commandeur JN (2010) Role of human glutathione S-transferases in the inactivation of reactive metabolites of clozapine. Chem Res Toxicol 23:1467–1476

    Article  CAS  Google Scholar 

  • Dragovic S, Gunness P, Ingelman-Sundberg M, Vermeulen NP, Commandeur JN (2013) Characterization of human cytochrome P450s involved in the bioactivation of clozapine. Drug Metab Dispos 41:651–658

    Article  CAS  Google Scholar 

  • Dur A, Kose H, Kocyigit A, Kocaman O, Ismayilova M, Sonmez FC (2016) The anti-inflammatory and antioxidant effects of thymoquinone on ceruleine induced acute pancreatitis in rats. Bratisl Lek Listy 117:614–618

  • Farkhondeh T, Samarghandian S, Borji A (2017) An overview on cardioprotective and anti-diabetic effects of thymoquinone. Asian Pac J Trop Med 10:849–854

    Article  CAS  Google Scholar 

  • Khalife K, Lupidi G (2008) Reduction of hypervalent states of myoglobin and hemoglobin to their ferrous forms by thymoquinone the role of GSH, NADH and NADPH. Biochim Biophys Acta 1780:627–637

    Article  CAS  Google Scholar 

  • Khezri S, Sabzalipour T, Jahedsani A, Azizian S, Atashbar S, Salimi A (2020) Chrysin ameliorates aluminum phosphide-induced oxidative stress and mitochondrial damages in rat cardiomyocytes and isolated mitochondria. Environ Toxicol 35:1114–1124

  • Kilian JG, Kerr K, Lawrence C, Celermajer DS (1999) Myocarditis and cardiomyopathy associated with clozapine. The Lancet 354:1841–1845

    Article  CAS  Google Scholar 

  • Liu H, Liu HY, Jiang YN, Li N (2016) Protective effect of thymoquinone improves cardiovascular function, and attenuates oxidative stress, inflammation and apoptosis by mediating the PI3K/Akt pathway in diabetic rats. Mol Med Rep 13:2836–2842

    Article  CAS  Google Scholar 

  • Longhi S, Heres S (2017) Clozapine-induced, dilated cardiomyopathy a case report. BMC Res Notes 10:1–3

    Article  Google Scholar 

  • McGill MR, Jaeschke H (2013) Metabolism and disposition of acetaminophen: recent advances in relation to hepatotoxicity and diagnosis. Pharm Res 30:2174–2187

    Article  CAS  Google Scholar 

  • Meltzer HY et al (2003) Clozapine treatment for suicidality in schizophrenia: international suicide prevention trial (InterSePT). Arch Gen Psychiatry 60:82–91

    Article  CAS  Google Scholar 

  • Nagi MN, Mansour MA (2000) Protective effect of thymoquinone against doxorubicin–induced cardiotoxicity in rats: a possible mechanism of protection. Pharmacol Res 41:283–289

    Article  CAS  Google Scholar 

  • Odinokova I et al (2018) Effect of melatonin on rat heart mitochondria in acute heart failure in aged rats. Int J Mol Sciences 19:1555

    Article  CAS  Google Scholar 

  • Orhan H (2015) Extrahepatic targets and cellular reactivity of drug metabolites. Curr Med Chem 22:408–437

    Article  CAS  Google Scholar 

  • Salimi A, Motallebi A, Ayatollahi M, Seydi E, Mohseni AR, Nazemi M, Pourahmad J (2017) Selective toxicity of persian gulf sea cucumber holothuria parva on human chronic lymphocytic leukemia b lymphocytes by direct mitochondrial targeting. Environ Toxicol 32:1158–1169

    Article  CAS  Google Scholar 

  • Samarghandian S, Azimini-Nezhad M, Farkhondeh T (2016) The effects of Zataria multiflora on blood glucose, lipid profile and oxidative stress parameters in adult mice during exposure to bisphenol. Cardiovasc Hematol Disord Drug Targets 16:41–46

    Article  CAS  Google Scholar 

  • Shi W, Deng H, Zhang J, Zhang Y, Zhang X, Cui G (2018) Mitochondria-targeting small molecules effectively prevent cardiotoxicity induced by doxorubicin. Molecules 23:1486

    Article  CAS  Google Scholar 

  • Shukla SK, Gupta S, Ojha SK, Sharma SB (2010) Cardiovascular friendly natural products: a promising approach in the management of CVD. Nat Prod Res 24:873–898

    Article  CAS  Google Scholar 

  • Silva SG, F, F Simoes R, Couto R, J Oliveira P, (2016) Targeting mitochondria in cardiovascular diseases. Curr Pharm Des 22:5698–5717

    Article  CAS  Google Scholar 

  • Stelmashook E, Chetverikov N, Golyshev S, Genrikhs E, Isaev N (2020) Thymoquinone induces mitochondrial damage and death of cerebellar granule neurons. Biochemistry (Moscow) 85:205–212

    Article  CAS  Google Scholar 

  • Williams DP, O’Donnell CJ, Maggs JL, Leeder JS, Uetrecht J, Pirmohamed M, Park BK (2003) Bioactivation of clozapine by murine cardiac tissue in vivo and in vitro. Chem Res Toxicol 16:1359–1364

    Article  CAS  Google Scholar 

Download references

Funding

This study was supported by Shahid Beheshti University of Medical Sciences, Deputy of Research.

Author information

Authors and Affiliations

  1. Cancer Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Asghar Ashrafi Hafez

  2. Student Research Committee, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran

    Zhaleh Jamali

  3. Department of Addiction Studies, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran

    Zhaleh Jamali

  4. Students Research Committee, School of Pharmacy, Ardabil University of Medical Sciences, Ardabil, Iran

    Saleh Khezri

  5. Traditional Medicine and Hydrotherapy Research Center, Ardabil University of Medical Sciences, Ardabil, Iran

    Saleh Khezri & Ahmad Salimi

  6. Department of Pharmacology and Toxicology, School of Pharmacy, Ardabil University of Medical Sciences, P.O. Box, 56189-53141, Ardabil, Iran

    Ahmad Salimi

Authors
  1. Asghar Ashrafi Hafez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhaleh Jamali
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Saleh Khezri
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ahmad Salimi
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

A.A. and A.S. conceived and designed research. S.K. and Z.J. conducted experiments. A.S. contributed new reagents or analytical tools. A.S. analyzed data. A.S. and A.A. wrote the manuscript. All authors read and approved the manuscript and all data were generated in-house and that no paper mill was used.

Corresponding author

Correspondence to Ahmad Salimi.

Ethics declarations

Ethical approval

This research was approved by the Ethics Committee at the Shahid Beheshti University of Medical Sciences, Tehran, Iran. The approval code is IR.SBMU.RETECH.REC.1399.494.

Consent to participate

All authors have given consent to their contribution.

Consent to publish

All authors have agreed with the content and all have given explicit consent to publish.

Competing interests

The authors declare no competing interests.

Additional information

Publisher's note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hafez, A.A., Jamali, Z., Khezri, S. et al. Thymoquinone reduces mitochondrial damage and death of cardiomyocytes induced by clozapine. Naunyn-Schmiedeberg's Arch Pharmacol 394, 1675–1684 (2021). https://doi.org/10.1007/s00210-021-02095-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00210-021-02095-1

Keywords

Search

Navigation