The Effect of Thymoquinone on Progressive Brain Damage: An Experimental Study in a Wistar Rat Model of Intracerebral Hemorrhage
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Submitted
12 September 2025
Published
30 June 2026
Keywords:
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MMP-9, Thymoquinone, Inflammatory, Free radicals, Intracerebral hemorrhage
Abstract
Intracerebral hemorrhage (ICH) accounts for 20% of stroke cases and is associated with a 40–50% mortality rate, yet effective neuroprotective therapies remain lacking. This study evaluated the therapeutic efficacy and molecular mechanisms of thymoquinone (TQ), the primary bioactive constituent of Nigella sativa, in a Wistar rat model of ICH. Male Wistar rats (n = 55, 200–220 g) were randomized into five groups: untreated control, untreated ICH, vehicle-treated ICH (corn oil for 7 days), and ICH treated orally with TQ at 150 mg/kg or 250 mg/kg body weight daily for 7 days. ICH was induced via autologous blood injection (0.12 mL) into the brain parenchyma. Biomarkers for neuroinflammation (NLRP3, TNF-α, IL-6, IL-1β), oxidative stress (SOD, MDA), tissue remodeling (MMP-9), and neuronal necrosis were quantified using ELISA, immunohistochemistry, and histological staining. TQ administration significantly elevated MMP-9 levels (p = 0.000) compared with untreated ICH controls and markedly suppressed NLRP3, TNF-α, and IL-1β expression. Furthermore, TQ increased SOD antioxidant activity, whereas MDA levels remained elevated. These neuroprotective effects were dose-dependent, with maximal modulation observed at 250 mg/kg TQ. In conclusion, TQ modulates critical inflammatory and oxidative pathways in experimental ICH while unexpectedly upregulating MMP-9, suggesting a complex dual function in early neuroprotection and subsequent tissue remodeling. These findings demonstrate that TQ holds promising potential as a natural neuroprotective agent for hemorrhagic stroke management.
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References
1. Montaño A, Hanley DF, Hemphill JC 3rd. Hemorrhagic stroke. Handb Clin Neurol. 2021;176:229-48. DOI: 10.1016/B978-0-444-64034-5.00019-5; PMID: 33272397.
2. Wang S, Zou XL, Wu LX, Zhou HF, Xiao L, Yao T, et al. Epidemiology of intracerebral hemorrhage: A systematic review and meta-analysis. Front Neurol. 2022;13:915813. DOI: 10.3389/fneur.2022.915813; PMID: 36188383; PMCID: PMC9523083.
3. Lindsay MP, Norrving B, Sacco RL, Brainin M, Hacke W, Martins S, et al. World Stroke Organization (WSO): Global Stroke Fact Sheet 2019. Int J Stroke. 2019;14(8):806-17. DOI: 10.1177/1747493019881353; PMID: 31658892.
4. Yudawijaya A, Suling FRW. Comparison of hypertension risk factors in Hemorric Stroke with Non-Hemorric Stroke in UKI General Hospital, East Jakarta. Int J Med Health Res. 2022;8(3):40-8.
5. Yuniati D, Syifak S, Putra PA, Saffanah VSP. Intracerebral hemorrhage Score as a Prognosis Prediction of Spontaneous Intracerebral hemorrhage at RSI Surabaya Jemursari. AKSONA. 2023;3(2):67-73. DOI: 10.20473/aksona.v3i2.40242.
6. Shao Z, Tu S, Shao A. Pathophysiological Mechanisms and Potential Therapeutic Targets in Intracerebral Hemorrhage. Front Pharmacol. 2019;10:1079. DOI: 10.3389/fphar.2019.01079; PMID: 31607923; PMCID: PMC6761372.
7. Zheng S, Jian D, Gan H, Wang L, Zhao J, Zhai X. FUNDC1 inhibits NLRP3-mediated inflammation after intracerebral hemorrhage by promoting mitophagy in mice. Neurosci Lett. 2021;756:135967. DOI: 10.1016/j.neulet.2021.135967; PMID: 34022268.
8. Liu T, Li X, Cui Y, Meng P, Zeng G, Wang Y, Wang Q. Bioinformatics Analysis Identifies Potential Ferroptosis Key Genes in the Pathogenesis of Intracerebral Hemorrhage. Front Neurosci. 2021;15:661663. DOI: 10.3389/fnins.2021.661663; PMID: 34163322; PMCID: PMC8215678.
9. Shad KF, Soubra W, Cordato DJ. The role of thymoquinone, a major constituent of Nigella sativa, in the treatment of inflammatory and infectious diseases. Clin Exp Pharmacol Physiol. 2021;48(11):1445-53. DOI: 10.1111/1440-1681.13553; PMID: 34297870.
10. Bordoni L, Fedeli D, Nasuti C, Maggi F, Papa F, Wabitsch M, De Caterina R, Gabbianelli R. Antioxidant and Anti-Inflammatory Properties of Nigella sativa Oil in Human Pre-Adipocytes. Antioxidants. 2019;8(2):51. DOI: 10.3390/antiox8020051; PMID: 30823525; PMCID: PMC6406245.
11. Tadi P, Lui F. Acute Stroke. In: StatPearls. Treasure Island (FL): StatPearls Publishing; 2026. NBKID: NBK535369
12. Amartey J, Gapper S, Hussein N, Morris K, Withycombe CE. Nigella sativa extract and thymoquinone regulate inflammatory cytokine and TET-2 expression in endothelial cells. Artery Res. 2019;25(3):157-63. DOI: 10.2991/artres.k.191114.002.
13. Cobourne-Duval MK, Taka E, Mendonca P, Soliman KFA. Thymoquinone increases the expression of neuroprotective proteins while decreasing the expression of pro-inflammatory cytokines and the gene expression NFκB pathway signaling targets in LPS/IFNγ -activated BV-2 microglia cells. J Neuroimmunol. 2018;320:87-97. DOI: 10.1016/j.jneuroim.2018.04.018; PMID: 29759145; PMCID: PMC5967628.
14. Durak MA, Ozhan O, Yildiz A, Durhan M, Vardi N, Cigremis Y, et al. Protective effect of short-term thymoquinone administration on the central nervous system in cisplatin-induced neurotoxicity. Eur Rev Med Pharmacol Sci. 2022;26(19):6935-43. DOI: 10.26355/eurrev_202210_29874; PMID: 36263573.
15. Hu X, Tao C, Gan Q, Zheng J, Li H, You C. Oxidative Stress in Intracerebral Hemorrhage: Sources, Mechanisms, and Therapeutic Targets. Oxid Med Cell Longev. 2016;2016:3215391. DOI: 10.1155/2016/3215391; PMID: 26843907; PMCID: PMC4710930.
16. Zhao S, Jiang J, Jing Y, Liu W, Yang X, Hou X, et al. The concentration of tumor necrosis factor-α determines its protective or damaging effect on liver injury by regulating Yap activity. Cell Death Dis. 2020;11(1):70. DOI: 10.1038/s41419-020-2264-z; PMID: 31988281; PMCID: PMC6985193.
17. Isaev NK, Genrikhs EE, Stelmashook EV. Antioxidant Thymoquinone and Its Potential in the Treatment of Neurological Diseases. Antioxidants. 2023;12(2):433. DOI: 10.3390/antiox12020433; PMID: 36829993; PMCID: PMC9952318.
18. Liu Y, Huang L, Kim MY, Cho JY. The Role of Thymoquinone in Inflammatory Response in Chronic Diseases. Int J Mol Sci. 2022;23(18):10246. DOI: 10.3390/ijms231810246; PMID: 36142148; PMCID: PMC9499585.
19. Fanoudi S, Alavi MS, Hosseini M, Sadeghnia HR. Nigella sativa and thymoquinone attenuate oxidative stress and cognitive impairment following cerebral hypoperfusion in rats. Metab Brain Dis. 2019;34(4):1001-10. DOI: 10.1007/s11011-019-00394-4; PMID: 31016464.
20. Könnecke H, Bechmann I. The role of microglia and matrix metalloproteinases involvement in neuroinflammation and gliomas. Clin Dev Immunol. 2013;2013:914104. DOI: 10.1155/2013/914104; PMID: 24023566; PMCID: PMC3759277.
21. Guan X, Li M, Li H, Guo Z, Ullah MS, Liu X, Wu M, Yu W. The role of MMPs in intracerebral hemorrhage. Front Cell Dev Biol. 2025;13:1667228. DOI: 10.3389/fcell.2025.1667228; PMID: 41480312; PMCID: PMC12754015.
22. Kaymak E, Akin AT, Öztürk E, Karabulut D, Kuloğlu N, Yakan B. Thymoquinone has a neuroprotective effect against inflammation, oxidative stress, and endoplasmic reticulum stress in the brain cortex, medulla, and hippocampus due to doxorubicin. J Biochem Mol Toxicol. 2021;35(11):e22888. DOI: 10.1002/jbt.22888; PMID: 34392583.
23. Behairy A, Elkomy A, Elsayed F, Gaballa MMS, Soliman A, Aboubakr M. Antioxidant and anti-inflammatory potential of spirulina and thymoquinone mitigate the methotrexate-induced neurotoxicity. Naunyn Schmiedebergs Arch Pharmacol. 2024;397(3):1875-88. DOI: 10.1007/s00210-023-02739-4; PMID: 37773524; PMCID: PMC10858838.
24. Zhang Y, Yu W, Liu Y, Chang W, Wang M, Zhang L. Regulation of nuclear factor erythroid-2-related factor 2 as a potential therapeutic target in intracerebral hemorrhage. Front Mol Neurosci. 2022;15:995518. DOI: 10.3389/fnmol.2022.995518; PMID: 36245922; PMCID: PMC9559574.
25. Pottoo FH, Ibrahim AM, Alammar A, Alsinan R, Aleid M, Alshehhi A, Alshehri M, Mishra S, Alhajri N. Thymoquinone: Review of Its Potential in the Treatment of Neurological Diseases. Pharmaceuticals. 2022;15(4):408. DOI: 10.3390/ph15040408; PMID: 35455405; PMCID: PMC9026861.
2. Wang S, Zou XL, Wu LX, Zhou HF, Xiao L, Yao T, et al. Epidemiology of intracerebral hemorrhage: A systematic review and meta-analysis. Front Neurol. 2022;13:915813. DOI: 10.3389/fneur.2022.915813; PMID: 36188383; PMCID: PMC9523083.
3. Lindsay MP, Norrving B, Sacco RL, Brainin M, Hacke W, Martins S, et al. World Stroke Organization (WSO): Global Stroke Fact Sheet 2019. Int J Stroke. 2019;14(8):806-17. DOI: 10.1177/1747493019881353; PMID: 31658892.
4. Yudawijaya A, Suling FRW. Comparison of hypertension risk factors in Hemorric Stroke with Non-Hemorric Stroke in UKI General Hospital, East Jakarta. Int J Med Health Res. 2022;8(3):40-8.
5. Yuniati D, Syifak S, Putra PA, Saffanah VSP. Intracerebral hemorrhage Score as a Prognosis Prediction of Spontaneous Intracerebral hemorrhage at RSI Surabaya Jemursari. AKSONA. 2023;3(2):67-73. DOI: 10.20473/aksona.v3i2.40242.
6. Shao Z, Tu S, Shao A. Pathophysiological Mechanisms and Potential Therapeutic Targets in Intracerebral Hemorrhage. Front Pharmacol. 2019;10:1079. DOI: 10.3389/fphar.2019.01079; PMID: 31607923; PMCID: PMC6761372.
7. Zheng S, Jian D, Gan H, Wang L, Zhao J, Zhai X. FUNDC1 inhibits NLRP3-mediated inflammation after intracerebral hemorrhage by promoting mitophagy in mice. Neurosci Lett. 2021;756:135967. DOI: 10.1016/j.neulet.2021.135967; PMID: 34022268.
8. Liu T, Li X, Cui Y, Meng P, Zeng G, Wang Y, Wang Q. Bioinformatics Analysis Identifies Potential Ferroptosis Key Genes in the Pathogenesis of Intracerebral Hemorrhage. Front Neurosci. 2021;15:661663. DOI: 10.3389/fnins.2021.661663; PMID: 34163322; PMCID: PMC8215678.
9. Shad KF, Soubra W, Cordato DJ. The role of thymoquinone, a major constituent of Nigella sativa, in the treatment of inflammatory and infectious diseases. Clin Exp Pharmacol Physiol. 2021;48(11):1445-53. DOI: 10.1111/1440-1681.13553; PMID: 34297870.
10. Bordoni L, Fedeli D, Nasuti C, Maggi F, Papa F, Wabitsch M, De Caterina R, Gabbianelli R. Antioxidant and Anti-Inflammatory Properties of Nigella sativa Oil in Human Pre-Adipocytes. Antioxidants. 2019;8(2):51. DOI: 10.3390/antiox8020051; PMID: 30823525; PMCID: PMC6406245.
11. Tadi P, Lui F. Acute Stroke. In: StatPearls. Treasure Island (FL): StatPearls Publishing; 2026. NBKID: NBK535369
12. Amartey J, Gapper S, Hussein N, Morris K, Withycombe CE. Nigella sativa extract and thymoquinone regulate inflammatory cytokine and TET-2 expression in endothelial cells. Artery Res. 2019;25(3):157-63. DOI: 10.2991/artres.k.191114.002.
13. Cobourne-Duval MK, Taka E, Mendonca P, Soliman KFA. Thymoquinone increases the expression of neuroprotective proteins while decreasing the expression of pro-inflammatory cytokines and the gene expression NFκB pathway signaling targets in LPS/IFNγ -activated BV-2 microglia cells. J Neuroimmunol. 2018;320:87-97. DOI: 10.1016/j.jneuroim.2018.04.018; PMID: 29759145; PMCID: PMC5967628.
14. Durak MA, Ozhan O, Yildiz A, Durhan M, Vardi N, Cigremis Y, et al. Protective effect of short-term thymoquinone administration on the central nervous system in cisplatin-induced neurotoxicity. Eur Rev Med Pharmacol Sci. 2022;26(19):6935-43. DOI: 10.26355/eurrev_202210_29874; PMID: 36263573.
15. Hu X, Tao C, Gan Q, Zheng J, Li H, You C. Oxidative Stress in Intracerebral Hemorrhage: Sources, Mechanisms, and Therapeutic Targets. Oxid Med Cell Longev. 2016;2016:3215391. DOI: 10.1155/2016/3215391; PMID: 26843907; PMCID: PMC4710930.
16. Zhao S, Jiang J, Jing Y, Liu W, Yang X, Hou X, et al. The concentration of tumor necrosis factor-α determines its protective or damaging effect on liver injury by regulating Yap activity. Cell Death Dis. 2020;11(1):70. DOI: 10.1038/s41419-020-2264-z; PMID: 31988281; PMCID: PMC6985193.
17. Isaev NK, Genrikhs EE, Stelmashook EV. Antioxidant Thymoquinone and Its Potential in the Treatment of Neurological Diseases. Antioxidants. 2023;12(2):433. DOI: 10.3390/antiox12020433; PMID: 36829993; PMCID: PMC9952318.
18. Liu Y, Huang L, Kim MY, Cho JY. The Role of Thymoquinone in Inflammatory Response in Chronic Diseases. Int J Mol Sci. 2022;23(18):10246. DOI: 10.3390/ijms231810246; PMID: 36142148; PMCID: PMC9499585.
19. Fanoudi S, Alavi MS, Hosseini M, Sadeghnia HR. Nigella sativa and thymoquinone attenuate oxidative stress and cognitive impairment following cerebral hypoperfusion in rats. Metab Brain Dis. 2019;34(4):1001-10. DOI: 10.1007/s11011-019-00394-4; PMID: 31016464.
20. Könnecke H, Bechmann I. The role of microglia and matrix metalloproteinases involvement in neuroinflammation and gliomas. Clin Dev Immunol. 2013;2013:914104. DOI: 10.1155/2013/914104; PMID: 24023566; PMCID: PMC3759277.
21. Guan X, Li M, Li H, Guo Z, Ullah MS, Liu X, Wu M, Yu W. The role of MMPs in intracerebral hemorrhage. Front Cell Dev Biol. 2025;13:1667228. DOI: 10.3389/fcell.2025.1667228; PMID: 41480312; PMCID: PMC12754015.
22. Kaymak E, Akin AT, Öztürk E, Karabulut D, Kuloğlu N, Yakan B. Thymoquinone has a neuroprotective effect against inflammation, oxidative stress, and endoplasmic reticulum stress in the brain cortex, medulla, and hippocampus due to doxorubicin. J Biochem Mol Toxicol. 2021;35(11):e22888. DOI: 10.1002/jbt.22888; PMID: 34392583.
23. Behairy A, Elkomy A, Elsayed F, Gaballa MMS, Soliman A, Aboubakr M. Antioxidant and anti-inflammatory potential of spirulina and thymoquinone mitigate the methotrexate-induced neurotoxicity. Naunyn Schmiedebergs Arch Pharmacol. 2024;397(3):1875-88. DOI: 10.1007/s00210-023-02739-4; PMID: 37773524; PMCID: PMC10858838.
24. Zhang Y, Yu W, Liu Y, Chang W, Wang M, Zhang L. Regulation of nuclear factor erythroid-2-related factor 2 as a potential therapeutic target in intracerebral hemorrhage. Front Mol Neurosci. 2022;15:995518. DOI: 10.3389/fnmol.2022.995518; PMID: 36245922; PMCID: PMC9559574.
25. Pottoo FH, Ibrahim AM, Alammar A, Alsinan R, Aleid M, Alshehhi A, Alshehri M, Mishra S, Alhajri N. Thymoquinone: Review of Its Potential in the Treatment of Neurological Diseases. Pharmaceuticals. 2022;15(4):408. DOI: 10.3390/ph15040408; PMID: 35455405; PMCID: PMC9026861.
Authors
1.
Thohari K, Fauzi AA, Purwanto DA. The Effect of Thymoquinone on Progressive Brain Damage: An Experimental Study in a Wistar Rat Model of Intracerebral Hemorrhage. Borneo J Pharm [Internet]. 2026Jun.30 [cited 2026Aug.14];9(2):153-64. Available from: https://journal.umpr.ac.id/index.php/bjop/article/view/10849
Copyright (c) 2026 Khamim Thohari, Asra Al Fauzi, Djoko Agus Purwanto
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