Document Type : Original Article


Department of Biochemistry, Ataturk University Faculty of Veterinary, Erzurum, Türkiye


Ischemia-reperfusion (IR) injury to the lower extremities causes damage to various tissues, notably the limbs. Because research in recent years have demonstrated that saffron and its components are useful in ischemic strokes, the goal of this study was to see whether Crocin (Cr), one of the active constituents in saffron, could protect the gastrocnemius muscle from IR injury. A total number of 32 Sprague-Dawley rats were randomized into four groups randomly: control, Cr, IR, and IR + Cr. Xylazine and ketamine were used to anesthetize all of the rats. The left lower limbs of the other two groups were subjected to 2 hr of ischemia and 2 hr of reperfusion with tourniquet, with the exception of the control and Cr groups. Tumor necrosis factor alfa (TNF-α), interleukin 6 (IL-6), IL-1β, total antioxidant status (TAS) and total oxidant status (TOS) levels were assessed in the blood as well as muscle IL-6, IL1β, SOD1-2, catalase (CAT) and glutathione peroxidase (GPx) expression. According to the IR group, increases in TAS levels and decreases in TNF-α, IL-6, and IL-1β levels were substantial in the Cr therapy group. Cr significantly reduced IL-6 and IL-1β mRNA expression levels in the muscle of the IR group and increased superoxide dismutases 1 (SOD1), SOD2, catalase (CAT), and GPx. Our data showed that Cr protected the gastrocnemius muscle from IR injury in rats and reduced inflammatory markers significantly. These effects of Cr might have been mediated by improved antioxidant enzyme activity, suppression of free radical generation and reduction of oxidative stress.


  1. Amani H, Mostafavi E, Alebouyeh MR, et al. Would colloidal gold nanocarriers present an effective diagnosis or treatment for ischemic stroke? Int J Nanomedicine 2019; 14: 8013-8031.
  2. Kumar V, Abbas AK, Aster JC. Robbins basic pathology. E-Book. 10th Amsterdam, Netherlands: Elsevier Health Sciences 2017; 57-97.
  3. Kraiss LW, Conte MS, Geary RL, et al. Setting high-impact clinical research priorities for the Society for Vascular Surgery. J Vasc Surg 2013; 57(2): 493-500. 
  4. Lejay A, Meyer A, Schlagowski AI, et al. Mitochondria: mitochondrial participation in ischemia-reperfusion injury in skeletal muscle. Int J Biochem Cell Biol 2014; 50: 101-105.
  5. Khayatnouri M, Safavi SE, Safarmashaei S, et al. The effect of saffron orally administration on spermato-genesis index in rat. Adv Environ Biol 2011; 5(7): 1514-1521.
  6. Assimopoulou AN, Sinakos Z, Papageorgiou VP. Radical scavenging activity of Crocus sativus L. extract and its bioactive constituents. Phytother Res 2005; 19(11): 997-
  7. Fernández JA. Anticancer properties of saffron, Crocus sativus Adv Phytomedicine 2006; 2: 313-330.
  8. Schmidt M, Betti G, Hensel A. Saffron in phytotherapy: pharmacology and clinical uses. Wien Med Wochenschr 2007; 157(13-14): 315-319.
  9. Hosseinzadeh H, Shamsaie F, Mehri S. Antioxidant activity of aqueous and ethanolic extracts of Crocus sativus stigma and its bioactive constituent, crocin and safranal. Pharmacogn Mag 2009; 5(20): 419-424.
  10. Wang Y, Han T, Zhu Y, et al. Antidepressant properties of bioactive fractions from the extract of Crocus sativus J Nat Med 2010; 64(1): 24-30.
  11. Ghadrdoost B, Vafaei AA, Rashidy-Pour A, et al. Protective effects of saffron extract and its active constituent crocin against oxidative stress and spatial learning and memory deficits induced by chronic stress in Eur J Pharmacol 2011; 667(1-3): 222-229.
  12. Hosseinzadeh H, Sadeghnia HR, Ghaeni FA, et al. Effects of saffron (Crocus sativus) and its active constituent, crocin, on recognition and spatial memory after chronic cerebral hypoperfusion in rats. Phytother Res 2012; 26(3): 381-386.
  13. Morelli S, Salerno S, Piscioneri A, et al. Neuronal membrane bioreactor as a tool for testing crocin neuroprotective effect in Alzheimer’s disease. Chem Eng J 2016; 305: 69-78.
  14. Wang C, Cai X, Hu W, et al. Investigation of the neuroprotective effects of crocin via antioxidant activities in HT22 cells and in mice with Alzheimer’s disease. Int J Mol Med 2019; 43(2): 956-966.
  15. Apaydin Yildirim B, Albayrak S. Protective effects of crocin on experimental gastrocnemius muscle ischemia/ reperfusion model in rat. EPSTEM 2018; 3: 116-120.
  16. Pottecher J, Kindo M, Chamaraux‐Tran TN et al. Skeletal muscle ischemia–reperfusion injury and cyclosporine A in the aging rat. Fundam Clin Pharmacol 2016; 30(3): 216-225.
  17. Mard SA, Azad SM, Ahangarpoor A. Protective effect of crocin on gastric mucosal lesions induced by ischemia-reperfusion injury in rats. Iran J Pharm Res 2016; 15(Suppl): 93-99.
  18. Erel O. A new automated colorimetric method for measuring total oxidant status. Clin Biochem 2005; 38(12): 1103-1111.
  19. Coşan DT, Oner C, Soyocak A, et al. Inhibition of Kv 1.3 and Kv 10.1 voltage gated potassium channels role on oxidative stressin breast cancer.[Turkish] Dicle Med J 2017; 44(1): 43-50.
  20. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 2001; 25(4): 402-408.
  21. Wang WZ, Fang XH, Stephenson LL, et al. Micro-circulatory effects of melatonin in rat skeletal muscle after prolonged ischemia. J Pineal Res 2005; 39(1): 57-65.
  22. Menger MD, Vollmar B. Pathomechanisms of ischemia-reperfusion injury as the basis for novel preventive strategies: is it time for the introduction of pleiotropic compounds? Transplant Proc 2007; 39(2): 485-488.
  23. Wang L, Shan Y, Chen L, et al. Colchicine protects rat skeletal muscle from ischemia/reperfusion injury by suppressing oxidative stress and inflammation. Iran J Basic Med Sci 2016; 19(6): 670-675.
  24. Chai H, Wang Q, Huang L, et al. Ginsenoside Rb1 inhibits tumor necrosis factor-alpha-induced vascular cell adhesion molecule-1 expression in human endo-thelial cells. Biol Pharm Bull 2008; 31(11): 2050-2056.
  25. Brüning CA, Prigol M, Luchese C, et al. Protective effect of diphenyl diselenide on ischemia and reperfusion-induced cerebral injury: involvement of oxidative stress and pro-inflammatory cytokines. Neurochem Res 2012; 37(10): 2249-2258.
  26. Oktar GL, Kirisci M, Dursun AD, et al. Antioxidative effects of adrenomedullin and vascular endothelial growth factor on lung injury induced by skeletal muscle ischemia-reperfusion. Bratisl Lek Listy 2013; 114(11): 625-628. 
  27. Khanna G, Diwan V, Singh M, et al. Reduction of ischemic, pharmacological and remote preconditioning effects by an antioxidant N-acetyl cysteine pretreat-ment in isolated rat heart. Yakugaku Zasshi 2008; 128(3): 469-477.
  28. Apaydin Yildirim B, Adoum BA. The investigation of the preventive effects of coenzyme Q10 and berberine for tourniquet induced ischemia-reperfusion injury on skeletal muscle in rat hindlimb. GSC Biol Pharm Sci 2019; 9(3): 127-133.
  29. Park UJ, Kim HT, Cho WH, et al. Remote ischemic preconditioning enhances the expression of genes encoding antioxidant enzymes and endoplasmic reticulum stress-related proteins in rat skeletal muscle. Vasc Specialist Int 2016; 32(4): 141-149.
  30. Mansour Z, Charles AL, Bouitbir J, et al. Remote and local ischemic postconditioning further impaired skeletal muscle mitochondrial function after ischemia-reperfusion. J Vasc Surg 2012; 56(3): 774-782.
  31. Ozyurt H, Ozyurt B, Koca K, et al. Caffeic acid phenethyl ester (CAPE) protects rat skeletal muscle against ischemia-reperfusion-induced oxidative stress. Vascul Pharmacol 2007; 47(2-3): 108-112.
  32. Kleikers PW, Wingler K, Hermans JJ, et al. NADPH oxidases as a source of oxidative stress and molecular target in ischemia/reperfusion injury. J Mol Med (Berl) 2012; 90(12): 1391-1406.