Histopathological and behavioral evaluations of the effects of crocin, safranal and insulin on diabetic peripheral neuropathy in rats

Document Type : Original Research Article


1 Division of Pathology, Department of Pathobiology, Faculty of Veterinary Medicine, Urmia University, Urmia, I.R. Iran

2 Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, I.R. Iran


Objectives: Crocin and safranal, the major constituents of saffron, exert neuroprotective effects. In the present study, we investigated the effects of crocin and safranal  (alone or in combination with insulin) on peripheral neuropathy in diabetic rats.
Materials and Methods: Diabetes was induced by intraperitoneal (i.p.) injection of 60 mg/kg of streptozotocin (STZ) and confirmed by blood glucose level higher than 250 mg/dl. After confirmation of diabetes, crocin (30 mg/kg, i.p.), safranal (1 mg/kg, i.p.) (alone or in combination with insulin) and insulin (5 IU/kg, s.c.) were administered for eight weeks. Neuropathic pain was evaluated using acetone drop test. Histopathological changes of sciatic nerve were evaluated using light microscope. Blood glucose levels and sciatic nerve malondialdehyde (MDA) contents were also measured.
Results: STZ caused cold allodynia, edema and degenerative changes of sciatic nerve, hyperglycemia and an elevation of sciatic nerve MDA levels. Crocin, safranal and insulin improved STZ-induced behavioral, histopathological and biochemical changes. Combined treatments produced more documented improving effects.
Conclusion: The results of the present study showed neuroprotective effects of crocin, safranal and insulin in a rat model of diabetic neuropathy. In addition, crocin and safranal enhanced the neuroprotective effect of insulin. The neuroprotective effects of theses chemical compounds could be associated with their anti-hyperglycemic and antioxidant properties.


Main Subjects

Alinejad B, Ghorbani A, Sadeghnia HR. 2013. Effects of combination of curcumin, linalool, rutin, safranal, and thymoquinone on glucose/serum deprivation-induced cell death. Avicenna J Phytomed, 3: 321-328.
Altinoz E, Oner Z, Elbe H, Cigremis Y, Turkoz Y. 2015. Protective effects of saffron (its active constituent, crocin) on nephropathy in streptozotocin-induced diabetic rats. Hum Exp Toxicol, 34: 127-134.
Asri-Rezaei S, tamaddonfard E, Ghasemsoltani-Momtaz B, Erfanparast A, Gholamalipour S. 2015. Effects of crocin and zinc chloride on blood levels of zinc and metabolic and oxidative parameters in streptozotocin-induced diabetic rats. Avicenna J Phytomed, 5: Article in Press.
Bujalska-Zadrozny M, de Carde A, Pawlik K. 2015. Influence of nitric oxide synthase or cyclooxygenase inhibitors on cannabinoids activity in streptozotocin-induced neuropathy. Pharm Rep, 67: 209-216.
Callaghan B, Cheng H, Stables C, Smith A, Felman F. 2012. Diabetic neuropathy: manifestations and current treatments. Lancet Neurol, 11: 251-234.
Choi Y, Yoon YM, Na HS, Kim SH, Chung JM. 1994. Behavioral signs of ongoing pain and cold allodynia in a rat model of neuropathic pain. Pain, 59: 369-376.
El-Abhar HS, Schaalan MF. 2014. Phytotherapy in diabetes: review on potential mechanistic perspectives. World J Diabetes, 5: 176-197.
England JD, Asbury AK. 2004. Peripheral neuropathy. Lancet, 363: 2151-2161.
Erfanparast A, Tamaddonfard E, Taati M, Dabbaghi M. 2015. Effects of crocin and safranal, saffron constituents, on the formalin-induced orofacial pain in rats. Avicenna J Phytomed, 5: Article in Press.
Erken HA, Genc O, Erken G, Ayada C, Gundagdu G, Dogan H. 2015. Ozone partially prevents diabetic neuropathy in rats. Exp Clin Endocrinol Diabetes, 123: 101-105.
Farshid AA, Tamaddonfard E, Najafi S. 2014. Effects of histidine and n-acetylcysteine on experimental lesions induced by doxorubicin in sciatic nerve of rats. Drug Chem Toxicol, DOI: 10.3109/014805545.2014981753.
Javed S, Petropoulos IN, Alam U, Malik RA. 2015. Treatment of painful diabetic neuropathy. Ther Adv Chronic Dis, 6: 15-28.
Karami M, Bathaie SZ, Tiraihi T, Habbibi-Rezaei M, Arabkheradmand J, Faghihzadeh S. 2013. Crocin improved locomotor function and mechanical behavior in a rat model of contused spinal cord injury through decreasing calcitonic gene related peptide (CGRP). Phytomedicine, 21: 62-67.
Kubo S, Watada H, Kawamori R. 2010. Combination therapy of miglitol and insulin in type 1 diabetes mellitus patients. Diabetes Investig, 22: 60-65.
Maeda A, kai K, Ishii M, Ishii T, Akagawa M. 2014. Safranal, a novel protein tyrosine phosphatase 1B inhibitor, activates insulin signaling in C2C12 myotubes and improved glucose tolerance in diabetic KK-Ay mice. Mol Nutr Food res, 58: 1177-1189.
Mielke JG, Wang YT. 2011. Insulin, synaptic function, and opportunities for neuroprotection. Prog Mol Biol Transl Sci, 98: 133-186.
Moore K, Roberts LJ. 1998. Measurement of lipid peroxidation. Free Radical Res, 28: 659-671.
Nam JS, Cheong YS, Karm MH, Ahn HS, Sim JH, Kim JS, Choi SS, Leem JG. 2014. Effects of nefopam on streptozotocin-induced diabetic neuropathic pain in rats. Korean J Pain, 27: 326-333.
Niswender KD. 2011. Basal insulin: beyond glycemia. Postgrad Med, 123: 27-37.
Ohkawa H, Oishi N, Tagi K. 1979. Assay for lipid peroxidase in animal tissues by thiobarbituric acid reaction. Anal Biochem, 95: 351-358.
Omran OM. 2012. Histopathological study of evening primrose oil effects on experimental diabetic neuropathy. Ultrustruct Pathol, 36: 222-227.
Patel DK, Prasad SK, Kumar R, Hemalatha S. 2012. An overview on antidiabetic medicinal plants having insulin mimetic property. Asian Pac J Trop Biomed, 2: 320-330.
Premkumar LS, Pabiddi RM. 2013. Diabetic peripheral neuropathy: role of reactive oxygen and nitrogen species. Cell Biochem Biophys, 67: 373-383.
Rajaei Z, Hadjzadeh MA, Nemati H, Hosseini M, Ahmadi M, Shafiee S. 2013. Antihyperglycemic and antioxidant activity of crocin in streptozotocin-induced diabetic rats. J Med Food, 16: 206-210.
Ranjitkumar R, Prathab Balaji S, Balaji B, Ramesh RV, Ramanathan M. 2013. Standardized aqueous Tribulus terristri (Nerunjil) extract attenuates hyperalgesia in experimentally induced diabetic neuropathic pain model: role of oxidative stress and inflammatory mediators. Phytother Res, 27: 1646-1657.
Sadeghnia HR, Kamkar M, Assadpour E, Boroushaki MT, Ghorbani A. 2013. Protective effect of safranal, a constituent of Crocus sativus, on quinolinic acid-induced oxidative damage in rat hippocampus. Iran J Basic Med Sci, 16: 73-82.
Samargandian S, Borji A, Delkhosh MB, Samini F. 2013. Safranal treatment improves hyperglycemia, hyperlipidemia and oxidative stress in streptozotocin-induced diabetic rats. J Pharm Pharm Sci, 16: 352-362.
Sharma S, Chopra K, Kulkarni SK. 2007. Effect of insulin and its combination with resveratrol or curcumin in attenuation of diabetic neuropathic pain: participation of nitric oxide and TNF-alpha. Phytother Res, 21: 278-283.
Singh R, Kishore L, Kaur N. 2014. Diabetic peripheral neuropathy: current perspectives and future directions. Pharmacol Res, 80: 21-35.
Tamaddonfard E, Farshid AA, Ahmadian E, Hamidhoseyni A. 2013a. Crocin enhanced functional recovery after sciatic nerve crush injury in rats. Iran J Basic Med Sci, 16: 83-90.
Tamaddonfard E, Farshid AA, Asri-Rezaee S, Javadi S, Khosravi V, Rahman B, Mirfakhraee Z. 2013b. Crocin improved learning and memory impairments in streptozotocin-induced diabetic rats. Iran J Basic Med Sci, 16: 91-100.
Tamaddonfard E, Farshid AA, Eghdami K, Samadi F, Erfanparst A. 2013c. Comparison of the effects of crocin, safranal and diclofenac on local inflammation and inflammatory pain responses induced by carrageenan in rats. Pharmacol Rep, 65: 1272-1280.
Tamaddonfard E, Farshid AA, Maroufi S, Kazemi-Shojaei S, Erfanparast A, Asri-Rezaei S, Taati M, Dabbaghi M, Escort M. 2014. Effects of safranal, a constituent of saffron, and vitamin E on nerve function and histopathology following crush injury of sciatic nerve in rats. Phytomedicine, 21: 717-723.
Tamaddonfard E, Hamzeh-Gooshchi N. 2010. Effect of crocin on the morphine-induced antinociception in the formalin test in rats. Phytother Res, 24: 410-413.
Tomlinson DR, Gardiner NJ. 2008. Glucose neurotoxicity. Nat Rev Neurosci, 9: 36-45.
Wayhs CA, Tortato C, Mescka CP, Pasquali MA, Schnorr CE, Nin MS, Barros HM, Moreira JC, Vargas CR. 2013. The association effect of insulin and clonazepam on oxidative stress in liver of an experimental animal model of diabetes and depression. Pharm Biol, 51: 533-538.
Wu YB, Shi LL, Wu YJ, Xu WH, Wang L, Ren MS. 2012. Protective effect of glidazide on diabetic peripheral neuropathy through Drp-1 mediated oxidative stress and apoptosis. Neurosci Lett, 523: 45-49.
Wu J, Yan LJ. 2015. Streptozotocin-induced type 1 diabetes in rodents as a model for studying mitochondrial mechanisms of diabetic β cell glucotoxicity. Diabetes Metab Syndr Obes, 8: 181-188.
Zhu KJ, Yang JS. 2014. Anti-allodynia effects of safranal on neuropathic pain induced by spinal nerve ligation in rats. Int J Clin Exp Med, 7: 4990-4996.
Zychowska M, Rojewska E, Przewlocka B, Mika J. 2013. Mechanisms and pharmacology of diabetic neuropathy-experimental and clinical studies. Pharmacol Rep, 65: 1601-1610.