Renoprotective effect of thymoquinone against rhabdomyolysis-induced acute kidney injury in the rat model

Document Type : Original Article


1 Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

2 Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

3 Department of Physiology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

4 Department of Pathology, Ghaem Hospital, Mashhad University of Medical Sciences, Mashhad, Iran


Objective(s): Rhabdomyolysis leads to the release of myoglobin, sarcoplasmic proteins, and electrolytes into the blood circulation causing acute kidney injury (AKI). Thymoquinone, a natural compound found in Nigella sativa seeds, has antioxidant and anti-inflammatory effects. This investigation assessed the renoprotective effect of thymoquinone on rhabdomyolysis-induced AKI in rats.
Materials and Methods: Male Wistar rats were categorized into six groups (n = 6): 1. Control: (normal saline), 2. Glycerol (50 ml/kg, single dose, IM), 3–5: Glycerol + thymoquinone (1, 2.5 and 5 mg/kg, 4 days, IP), 6. Thymoquinone (5 mg/kg). On day 5, serum and kidney tissue were isolated and the amounts of serum creatinine and blood urea nitrogen (BUN), renal malondialdehyde (MDA), glutathione (GSH.), tumor necrosis factor-alpha (TNF-α), neutrophil gelatinase-associated lipocalin (NGAL), and pathological changes were evaluated. 
Results: Glycerol increased creatinine, BUN, MDA, TNF-α, and NGAL levels. It decreased GSH amounts and caused renal tubular necrosis, glomerular atrophy, and myoglobin cast in kidney tissue. Co-administration of glycerol and thymoquinone reduced creatinine, BUN, histopathological alterations, and MDA levels, and enhanced GSH amounts. Administration of glycerol and thymoquinone (5 mg/kg) had no significant effect on TNF-α amount but decreased NGAL protein levels. The administration of thymoquinone (5 mg/kg) alone did not display a significant difference from the control group.
Conclusion: Rhabdomyolysis from glycerol injection in rats can cause kidney damage. Thymoquinone may attenuate renal dysfunction and oxidative stress. However, the TNF-α level was not significantly affected. Further studies are needed to explore the potential therapeutic effects of thymoquinone in managing AKI.


Main Subjects

1. Torres PA, Helmstetter JA, Kaye AM, Kaye AD. Rhabdomyolysis: pathogenesis, diagnosis, and treatment. Ochsner J 2015; 15: 58-69.
2. Petejova N, Martinek A. Acute kidney injury due to rhabdomyolysis and renal replacement therapy: A critical review. Crit Care 2014; 18: 224-231.
3. Boutaud O, Roberts II LJ. Mechanism-based therapeutic approaches to rhabdomyolysis-induced renal failure. Free Radic  Biol Med 2011; 51: 1062-1067.
4. Al-Brakati A, Alsharif KF, Alzahrani KJ, Kabrah S, Al-Amer O, Oyouni AA, et al. Using green biosynthesized lycopene-coated selenium nanoparticles to rescue renal damage in glycerol-induced acute kidney injury in rats. Int J Nanomed 2021; 16: 4335-4349.
5. Wu J, Pan X, Fu H, Zheng Y, Dai Y, Yin Y, et al. Effect of curcumin on glycerol-induced acute kidney injury in rats. Sci Rep 2017; 7: 1-11.
6. Reis NG, Francescato HDC, de Almeida LF, Silva C, Costa RS, Coimbra TM. Protective effect of calcitriol on rhabdomyolysis-induced acute kidney injury in rats. Sci Rep 2019; 9: 7090.
7. Yin M, Jiang N, Guo L, Ni Z, Al-Brakati AY, Othman MS, et al. Oleuropein suppresses oxidative, inflammatory, and apoptotic responses following glycerol-induced acute kidney injury in rats. Life Sci 2019; 232: 116634.
8. Boozari M, Hosseinzadeh H. Natural medicines for acute renal failure: A review. Phytother Res 2017; 31: 1824-1835.
9. Vafaeipour Z, Ghasemzadeh Rahbardar M, Hosseinzadeh H. Effect of saffron, black seed, and their main constituents on inflammatory cytokine response (mainly TNF-α) and oxidative stress status: An aspect on pharmacological insights. Naunyn Schmiedebergs Arch Pharmacol 2023; 396:2241-2259.
10. Oskouei Z, Akaberi M, Hosseinzadeh H. A glance at Black cumin (Nigella sativa) and its active constituent, thymoquinone, in ischemia: A review. Iran J Basic Med Sci 2018; 21: 1200-1209.
11. Hannan MA, Rahman MA, Sohag AAM, Uddin MJ, Dash R, Sikder MH, et al. Black Cumin (Nigella sativa L.): A comprehensive review on phytochemistry, health benefits, molecular pharmacology, and safety. Nutrients 2021; 13: 1784.
12. Nakisa N, Ghasemzadeh Rahbardar M. Action mechanisms of antirheumatic herbal medicines. Rheumatoid Arthritis: Intech Open 2021; 171-184.
13. Fadishei M, Ghasemzadeh Rahbardar M, Imenshahidi M, Mohajeri A, Razavi BM, Hosseinzadeh H. Effects of Nigella sativa oil and thymoquinone against bisphenol A-induced metabolic disorder in rats. Phytother Res 2021; 35: 2005-2024.
14. Hosseinzadeh H, Parvardeh S. Anticonvulsant effects of thymoquinone, the major constituent of Nigella sativa seeds, in mice. Phytomedicine 2004; 11: 56-64.
15. Oskouei Z, Mehri S, Kalalinia F, Hosseinzadeh H. Evaluation of the effect of thymoquinone in d-galactose-induced memory impairments in rats: Role of MAPK, oxidative stress, and neuroinflammation pathways and telomere length. Phytother Res 2021; 35: 2252-2266.
16. Turhan Caskurlu M. Mustafa Erboga, 3, Zeynep Fidanol Erboga MO, 4 Gokhan Atis1. Protective effect of Nigella sativa on renal reperfusion injury in rat. Iran J kidney Dis 2016; 10: 135-143.
17. Hammad FT, Lubbad L. The effect of thymoquinone on the renal functions following ischemia-reperfusion injury in the rat. Int J Physiol Pathophysiol Pharmacol 2016; 8: 152-159.
18. Farooqui Z, Ahmed F, Rizwan S, Shahid F, Khan AA, Khan F. Protective effect of Nigella sativa oil on cisplatin induced nephrotoxicity and oxidative damage in rat kidney. Biomed Pharmacother 2017; 85: 7-15.
19. Ahmad S, Beg ZH. Hypolipidemic and antioxidant activities of thymoquinone and limonene in atherogenic suspension fed rats. Food chem 2013; 138: 1116-1124.
20. Guan D, Li Y, Peng X, Zhao H, Mao Y, Cui Y. Thymoquinone protects against cerebral small vessel disease: Role of antioxidant and anti-inflammatory activities. J Biol Regul Homeostatic Agents 2018; 32: 225-231.
21. Amin B, Hosseinzadeh H. Black Cumin (Nigella sativa) and its active constituent, thymoquinone: An overview on the analgesic and anti-inflammatory effects. Planta Med 2016; 82: 8-16.
22. Hosseinzadeh H, Parvardeh S, Asl MN, Sadeghnia HR, Ziaee T. Effect of thymoquinone and Nigella sativa seeds oil on lipid peroxidation level during global cerebral ischemia-reperfusion injury in rat hippocampus. Phytomedicine 2007; 14: 621-627.
23. Calderone L, Grimes P, Shalev M. Acute reversible cataract induced by xylazine and by ketamine-xylazine anesthesia in rats and mice. Exp Eye Res 1986; 42: 331-337.
24. Liano F, Pascual J, Group MARFS. Epidemiology of acute renal failure: a prospective, multicenter, community-based study. Kidney Int 1996; 50: 811-818.
25. Mihara M, Uchiyama M. Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal Biochem 1978; 86: 271-278.
26. Ghasemzadeh Rahbardar M, Cheraghi Farmad H, Hosseinzadeh H, Mehri S. Protective effects of selenium on acrylamide-induced neurotoxicity and hepatotoxicity in rats. Iran J Basic Med Sci 2021; 24: 1041-1049.
27. Moron MS, Depierre JW, Mannervik B. Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim Biophys Acta 1979; 582: 67-78.
28. Ghasemzadeh Rahbardar M, Razavi BM, Hosseinzadeh H. Investigating the ameliorative effect of alpha-mangostin on development and existing pain in a rat model of neuropathic pain. Phytother Res 2020; 34: 3211-3225.
29. Ardakanian A, Ghasemzadeh Rahbardar M, Omidkhoda F, Razavi BM, Hosseinzadeh H. Effect of alpha-mangostin on olanzapine-induced metabolic disorders in rats. Iran J Basic Med Sci 2022; 25: 198-207.
30. Li Y-f, Xu B-y, An R, Du X-f, Yu K, Sun J-h, et al. Protective effect of anisodamine in rats with glycerol-induced acute kidney injury. BMC Nephrol 2019; 20: 1-14.
31. Hofni A, Ali FE, Ibrahim AR, Aboubaker EM. Renoprotective Effect of thymoquinone against streptozotocin-induced diabetic nephropathy: Role of NOX2 and Nrf2 signals. Curr Mol Pharmacol 2023; 16: 905-914.
32. Alsharidah M, Abdel-Moneim A-MH, Alsharidah AS, Mobark MA, Rahmani AH, Shata A, et al. Thymoquinone, but not metformin, protects against gentamicin-induced nephrotoxicity and renal dysfunction in rats. Appl Sci 2021; 11: 3981.
33. Mahmoud AM, Ahmed OM, Galaly SR. Thymoquinone and curcumin attenuate gentamicin-induced renal oxidative stress, inflammation and apoptosis in rats. Excli J 2014; 13: 98-110.
34. Nishida K, Watanabe H, Ogaki S, Kodama A, Tanaka R, Imafuku T, et al. Renoprotective effect of long acting thioredoxin by modulating oxidative stress and macrophage migration inhibitory factor against rhabdomyolysis-associated acute kidney injury. Sci Rep 2015; 5: 14471.
35. Homsi E, Andreazzi DD, Faria JB, Janino P. TNF-α-mediated cardiorenal injury after rhabdomyolysis in rats. Am J Physiol 2015; 308: 1259-1267.
36. Sharawy MH, Abdelrahman RS, El-Kashef DH. Agmatine attenuates rhabdomyolysis-induced acute kidney injury in rats in a dose dependent manner. Life Sci 2018; 208: 79-86.
37. Elsherbiny NM, Maysarah NM, El-Sherbiny M, Al-Gayyar MM. Renal protective effects of thymoquinone against sodium nitrite-induced chronic toxicity in rats: Impact on inflammation and apoptosis. Life Sci 2017; 180: 1-8.
38. Sener U, Uygur R, Aktas C, Uygur E, Erboga M, Balkas G, et al. Protective effects of thymoquinone against apoptosis and oxidative stress by arsenic in rat kidney. Ren Fail 2016; 38: 117-123.
39. He Z, Li J, Mei Y, Lv Y, Zhu F, Liu R, et al. The role and difference of TLR2 and TLR4 in rhabdomyolysis induced acute kidney injury in mice. Int J Clin Exp Pathol 2018; 11: 1054-1061.
40. Guo L-P, Liu S-X, Yang Q, Liu H-Y, Xu L-L, Hao Y-H, et al. Effect of thymoquinone on acute kidney injury induced by sepsis in BALB/c mice. Bio Med Res Int 2020; 2020.
41. Ali BH, Al Za’abi M, Shalaby A, Manoj P, Waly MI, Yasin J, et al. The effect of thymoquinone treatment on the combined renal and pulmonary toxicity of cisplatin and diesel exhaust particles. Exp Biol Med  2015; 240: 1698-1707.