Effect of thymoquinone, a constituent of Nigella sativa L., on ischemia–reperfusion in rat skeletal muscle

  • Hossein Hosseinzadeh
  • Samaneh Taiari
  • Marjan Nassiri-Asl
Original Article

Abstract

Thymoquinone have been shown to decrease ischemia–reperfusion injury (IRI) in some tissues such as gastric mucosa and brain. In this study, the effect of thymoquinone was evaluated on an animal model of IRI in the rat hind limb. Hind limb ischemia was induced by clamping the common femoral artery and vein. After 2 h ischemia, the clamp on the femoral vessels was taken off and the animal underwent 1 h reperfusion. Muscle injuries were evaluated by recording the electromyographic (EMG) potentials and performing some biochemical analysis including thiobarbituric acid reactive substances (TBARS), total sulfhydryl (SH) groups, and antioxidant capacity of muscle using ferric reducing ability of plasma (FRAP) assay. Ischemia was induced using free-flap surgery in skeletal muscle. Thymoquinone (20, 40 and 80 mg/kg) and normal saline (10 ml/kg) were administered intraperitoneally 1 h prior to reperfusion. The average peak-to-peak amplitude during ischemic reperfusion was significantly increased in thymoquinone groups in comparison with the control group. Following thymoquinone administration, the total SH contents and antioxidant capacity were elevated in muscle flap. The malondialdehyde (MDA) level was declined significantly in test groups. It is concluded that thymoquinone have some protective effects against the muscle tissue injury caused by lower limb ischemia–reperfusion.

Keywords

Thymoquinone Nigella sativa Oxidative stress Lower limb ischemia Reperfusion Electromyography 

Notes

Acknowledgements

The authors are thankful to the Vice Chancellor of Research, Mashhad University of Medical Sciences for financial support. The results described in this paper are part of a Pharm. D. thesis.

References

  1. Abdel-Fattah AFM, Matsumoto K, Watanabe H (2000) Antinociceptive effects of Nigella sativa oil and its major component, thymoquinone, in mice. Eur J Pharmacol 400:89–97PubMedCrossRefGoogle Scholar
  2. Abdollahi M, Ranjbar R, Shadnia S, Nikfar S, Rezaie A (2004) Pesticides and oxidative stress: a review. Med Sci Mon 10:141–147Google Scholar
  3. Al-Gharably NM, Badary OA, Nagi MN, Al-Sawaf HA, Al-Rikabi AC, Al-Bekairi AM (1997) Protective effect of thymoquinone against carbon tetrachloride-induced hepatotoxicity in mice. Res Commun Pharmacol Toxicol 2:41–50Google Scholar
  4. Awad AS, Kamel R, Sherief MA (2011) Effect of thymoquinone on hepatorenal dysfunction and alteration of CYP3A1 and spermidine/spermine N-1-acetyl-transferase gene expression induced by renal ischaemia–reperfusion in rats. J Pharm Pharmacol 63:1037–1042PubMedCrossRefGoogle Scholar
  5. Benzie IFF, Strain JJ (1996) The ferric reducing ability of plasma (FRAP) as a measure of antioxidant power: the FRAP assay. Anal Biochem 239:70–76PubMedCrossRefGoogle Scholar
  6. Benzie IFF, Strain JJ (1999) Ferric reducing/antioxidant power assay: direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration. Method Enzymol 299:15–27CrossRefGoogle Scholar
  7. Blaisdell FW (2002) The pathophysiology of skeletal muscle ischemia and the reperfusion syndrome: a review. Cardiovasc Surg 10:620–630PubMedCrossRefGoogle Scholar
  8. Burits M, Bucar F (2000) Antioxidant activity of Nigella sativa essential oil. Phytother Res 14:323–328PubMedCrossRefGoogle Scholar
  9. Daba MH, Abdel-Rahman MS (1998) Hepatoprotective activity of thymoquinone in isolated rat hepatocytes. Toxicol Lett 95:23–29PubMedCrossRefGoogle Scholar
  10. El-Abhar HS, Abdallah M, Saleh S (2003) Gastroprotective activity of Nigella sativa oil and its constituent, thymoquinone, against gastric mucosal injury induced by ischaemia–reperfusion in rats. J Ethnopharmacol 84:251–258PubMedCrossRefGoogle Scholar
  11. El-Dakhakhny M, Mady NI, Halim MA (2000) Nigella sativa L. protects against induced hepatotoxicity and improves serum lipid profile in rats. Arzneimittelforsch 50:832–836PubMedGoogle Scholar
  12. Ellman G (1959) Tissue sulfhydryl groups. Arch Biochem Biophys 82:70–77PubMedCrossRefGoogle Scholar
  13. Fernandez J, Perez-Alvarez JA, Fernandez-Lopez JA (1997) Thiobarbituric acid test for monitoring lipid oxidation in meat. Food Chem 99:345–353CrossRefGoogle Scholar
  14. Fernandez DC, Antuna SS, Martinez E, Arias AP (2000) Ischemia–reperfusion injury in muscular flaps: role of superoxide dismutase and catalase, 6th Internet World Congress for Biomedical Sciences, INABIS, Presentation# 159Google Scholar
  15. Grisotto PC, dos Santos AC, Coutinho-Neto J, Cherry J, Picinatto CE (2000) Indicators of oxidative injury and alterations of the cell membrane in the skeletal muscle of rats submitted to ischemia and reperfusion. J Surg Res 92:1–6PubMedCrossRefGoogle Scholar
  16. Gute DC, Ishida T, Yarimizu K, Korthuis RJ (1998) Inflammatory responses to ischemia and reperfusion in skeletal muscle. Mol Cell Biochem 179:169–187PubMedCrossRefGoogle Scholar
  17. Hamdy NM, Taha RA (2009) Effects of Nigella sativa oil and thymoquinone on oxidative stress and neuropathy in streptozotocin-induced diabetic rats. Pharmacology 84:127–134PubMedCrossRefGoogle Scholar
  18. Hassan M, El-Dakhakhny M (1992) Effect of some Nigella sativa constituents on chemical carcinogenesis in hamster cheek pouch. J Egypt Soc Pharmacol Exp Ther 11:675–677Google Scholar
  19. Hosseinzadeh H, Hosseini A, Nassiri-Asl M, Sadeghnia HR (2007a) Effect of Salvia leriifolia Benth. root extracts on ischemia–reperfusion in rat skeletal muscle. BMC Complement Alter Med 7:23CrossRefGoogle Scholar
  20. Hosseinzadeh H, Moghim FF, Mansouri SMT (2007b) Effect of Nigella sativa seed extracts on ischemia–reperfusion in rat skeletal muscle. Pharmacol gyonline 2:326–335Google Scholar
  21. Hosseinzadeh H, Parvardeh S, Asl MN, Sadeghnia HR, Ziaee T (2007c) Effect of thymoquinone and Nigella sativa seeds oil on lipid peroxidation level during global cerebral ischemia–reperfusion injury in rat hippocampus. Phytomedicine 14:621–627PubMedCrossRefGoogle Scholar
  22. Hosseinzadeh H, Modaghegh MH, Saffari Z (2009) Crocus sativus L. (saffron) extract and its active constituents (crocin and safranal) on ischemia–reperfusion in rat skeletal muscle. eCAM 6:343–350PubMedGoogle Scholar
  23. Houghton PJ, Zarka R, De-Las-Heras B, Hoult JR (1995) Fixed oil of Nigella sativa and derived thymoquinone inhibit Eicosanoid generation in leukocytes and membrane lipid peroxidation. Planta Med 61:33–36PubMedCrossRefGoogle Scholar
  24. Huang D, Ou B, Prior RL (2005) The chemistry behind antioxidant capacity assays. J Agric Food Chem 53:1841–1856PubMedCrossRefGoogle Scholar
  25. Iida H, Schmelzer JD, Schmeichel AM, Wang Y, Low PA (2003) Peripheral nerve ischemia: reperfusion injury and fiber regeneration. Exp Neurol 184:997–1002PubMedCrossRefGoogle Scholar
  26. Ismail M, Al-Naqeep G, Chan KW (2010) Nigella sativa thymoquinone-rich fraction greatly improves plasma antioxidant capacity and expression of antioxidant genes in hypercholesterolemic rats. Free Radic Biol Med 48:664–672PubMedCrossRefGoogle Scholar
  27. Korthals JK, Maki T, Gieron MA (1985) Nerve and muscle vulnerability to ischemia. J Neurol Sci 71:283–290PubMedCrossRefGoogle Scholar
  28. Mahgoub AA (2003) Thymoquinone protects against experimental colitis in rats. Toxicol Lett 143:133–143PubMedCrossRefGoogle Scholar
  29. Mansour MA, Nagi MN, El-Khatib AS, Al-Bekairi AM (2002) Effects of thymoquinone on antioxidant enzyme activities, lipid peroxidation and DT-diaphorase in different tissues of mice: a possible mechanism of action. Cell Biochem Funct 20:143–151PubMedCrossRefGoogle Scholar
  30. Mihara M, Uchiyama M (1978) Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal Biochem 86:271–278PubMedCrossRefGoogle Scholar
  31. Nagamatsu M, Schmelzer JD, Zollman PJ, Smithson IL, Nickander KK, Low PA (1996) Ischemic reperfusion causes lipid peroxidation and fiber degeneration. Muscle Nerve 19:37–47PubMedCrossRefGoogle Scholar
  32. Nagi MN, Alam K, Badary OA, Al-Shabaneh OA, Al-Sawaf HA, Al-Bekairi AM (1999) Thymoquinone protects against carbon tetrachloride hepatotoxicity in mice via an antioxidant mechanism. Biochem Mol Biol Int 47:153–159PubMedGoogle Scholar
  33. Oguzhanoglu A, Kurt T, Sahiner T (2000) Nerve conduction parameters during ischemia–reperfusion in the rat sciatic nerve. Electromyogr Clin Neurophysiol 40:487–490PubMedGoogle Scholar
  34. Ossowska K, Lorence-Koci E, Schulze G, Wolfarth S (1996) The influence of dizolcipine (MK-801) on the reserpine-enhanced electromyographic stretch reflex in rats. Neurosci Lett 203:73–76PubMedCrossRefGoogle Scholar
  35. Parihar MS, Hemnani T (2003) Phenolic antioxidants attenuate hippocampal neuronal cell damage against kainic acid-induced excitotoxicity. J Biosci 28:121–128PubMedCrossRefGoogle Scholar
  36. Schmelzer JD, Zochodne DW, Low PA (1989) Ischemic and reperfusion injury of rat peripheral nerve. Proc Natl Acad Sci USA 86:1639–1342PubMedCrossRefGoogle Scholar
  37. Soszynski M, Bartosz G (1997) Decrease in accessible thiols as an index of oxidative damage to membrane proteins. Free Rad Biol Med 23:463–469PubMedCrossRefGoogle Scholar
  38. Woo CC, Loo SY, Gee V, Yap CW, Sethi G, Kumar AP, Benny Tan KH (2011) Anticancer activity of thymoquinone in breast cancer cells: possible involvement of PPAR-γ pathway. Biochem Pharmacol. in pressGoogle Scholar
  39. Woodruff TM, Arumugam TV, Shiels IA, Reid RC, Fairlie DP, Taylor SM (2004) Protective effects of a potent C5a receptor antagonist on experimental acute limb ischemia–reperfusion in rats. J Surg Res 116:81–90PubMedCrossRefGoogle Scholar
  40. Worthen DR, Ghosheh OA, Crooks PA (1998) The in vitro anti-tumor activity of some crude and purified components of blackseed Nigella sativa L. Anticancer Res 18:1527–1532PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Hossein Hosseinzadeh
    • 1
  • Samaneh Taiari
    • 2
  • Marjan Nassiri-Asl
    • 3
  1. 1.Pharmaceutical Research Center, School of PharmacyMashhad University of Medical SciencesMashhadI.R. Iran
  2. 2.School of PharmacyMashhad University of Medical SciencesMashhadI. R. Iran
  3. 3.Department of Pharmacology, Faculty of MedicineQazvin University of Medical SciencesQazvinI.R. Iran

Personalised recommendations