Effect of Crocus sativus extracts and its active constituent safranal on the harmaline-induced tremor in mice

Document Type: Original Article


1 Department of Pharmacology and Physiology, Faculty of Medicine, Sabzevar University of Medical Sciences, Sabzevar, Iran

2 Neuroscience Research center, Department of Toxicology and Pharmacology, School of Pharmacy, Kerman University of Medical Sciences, Kerman, Iran

3 Pharmaceutical Research center, Department of Toxicology and Pharmacology, Faculty of Pharmacy, Kerman University of Medical Sciences, Kerman, Iran

4 Pharmaceutical Research Center, Department of Pharmacodynamics and Toxicology School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran


Objective(s): Due to unsatisfactory response or intolerable side effects of current drugs, treatment of essential tremor remains inadequate. Thus, we aimed to investigate the protective and therapeutic effects of aqueous and ethanolic extracts of Crocus sativus (saffron), and its active consistent, safranal, on the harmaline-induced tremor in mice.
Materials and Methods: To induce tremor, harmaline (30 mg/kg) was injected intraperitoneally. Test groups were also given the aqueous and ethanolic extracts of saffron (40, 80, and 160 mg/kg) as well as safranal (0.1, 0.3, and 0.5 ml/kg), intraperitoneally, 10 min before harmaline administration (prophylactic study) or 10 min after the onset of tremors (curative study). The latency of onset, duration, and intensity of tremor were recorded.
Results: The extracts (80 and160 mg/kg) dose dependently attenuated duration of harmaline-induced tremors as did reference drug, propranolol (2 and 5 mg/kg). Only the highest dose of extracts (160 mg/kg) attenuated intensity of harmaline-induced tremors throughout the study. Safranal at the doses of (0.1 and 0.3 ml/kg) but not 0.5 ml/kg attenuated duration and intensity of tremor. Onset of tremor increased with the extracts (80 and 160 mg/kg) in prophylactic study, as the effect observed with propranolol at the dose of 5 mg/kg. Safranal did not affect the latency of tremor.
Conclusion: Both aqueous and ethanolic extracts of saffron and with a less effect, low doses of safranal, have relatively protective and suppressive effects on the harmaline-induced tremor and different constituents of extracts seem to participate in the protective effects against harmaline induced tremor.


1. Elble R, Deuschl G. Milestones in tremor research. Mov Disord 2011; 26:1096-1105.

2. Zesiewicz TA, Elble RJ, Louis ED, Gronseth GS, Ondo WG, Dewey RB Jr, et al. Evidence-based guideline update: treatment of essential tremor: report of the quality standards subcommittee of the American Academy of Neurology. Neurology 2011; 77:1752-1755.

3. Deuschl G, Raethjen J, Hellriegel H, Elble R. Treatment of patients with essential tremor. Lancet Neurol 2011; 10:148-161.

4. Bernard JF, Horcholle-Bossavit G. Harmaline-induced rhythm in the lateral reticular nucleus. Arch Ital Biol 1983; 121:139-150.

5. Ahmed A, Taylor NR. The analysis of drug-induced tremor in mice. Br J Pharmacol Chemother 1959; 14:350-354.

6. Zargari A. Medicinal plants. Tehran: University Press; 1990:574-578.

7. Hosseinzadeh H, Nassiriā€Asl M. Avicenna's (Ibn Sina) the canon of medicine and saffron (Crocus sativus): a review. Phytother Res 2013; 27:475-483.

8. Abe K, Saito H. Effects of saffron extract and its constituent crocin on learning behaviour and long-term potentiation. Phytother Res 2000; 14:149-152.

9. Rastgoo M, Hosseinzadeh H, Alavizadeh H, Abbasi A, Ayati Z, Jaafari MR. Antitumor activity of PEGylated nanoliposomes containing crocin in mice bearing C26 colon carcinoma. Planta Med 2013; 79:447-451.

10. Hosseinzadeh H, Shamsaie F, Mehri S. Antioxidant activity of aqueous and ethanolic extracts of Crocus sativus L. stigma and its bioactive constituents, crocin and safranal. Pharmacogn Mag 2009; 5:419-424.

11. Razavi M, Hosseinzadeh H, Abnous K, Motamedshariaty VS, Imenshahidi M. Crocin restores hypotensive effect of subchronic administration of diazinon in rats. Iran J Basic Med Sci 2013; 16:64-72.

12. Mehdizadeh R, Parizadeh MR, Khooei AR, Mehri S, Hosseinzadeh H. Cardioprotective effect of saffron extract and safranal in isoproterenol-induced myocardial infarction in wistar rats. Iran J Basic Med Sci 2013; 16:56-63.

13. Hosseinzadeh H, Karimi G, Niapoor M. Antidepressant effect of Crocus sativus L. stigma extracts and their constituents, crocin and safranal, in mice. International Symposium on Saffron Biology and Biotechnology (ISHS) 2003; 650:435-445.

14. Wang Y, Han T, Zhu Y, Zheng CJ, Ming QL, Rahman K, et al. Antidepressant properties of bioactive fractions from the extract of Crocus sativus L. J Nat Med 2010; 64:24-30.

15. Hosseinzadeh H, Noraei NB. Anxiolytic and hypnotic effect of Crocus sativus aqueous extract and its constituents, crocin and safranal, in mice. Phytother Res 2009; 23:768-774.

16. Hosseinzadeh H, Ziaei T. Effects of Crocus sativus stigma extract and its constituents, crocin and safranal, on intact memory and scopolamine-induced learning deficits in rats performing the Morris water maze task. J Med Plants 2006; 5:40-50.

17. Hosseinzadeh H, Sadeghnia HR, Ghaeni FA, Motamedshariaty VS, Mohajeri SA. Effects of saffron (Crocus sativus L.) and its active constituent, crocin, on recognition and spatial memory after chronic cerebral hypoperfusion in rats. Phytother Res 2012; 26:381-386.

18. Hosseinzadeh H, Younesi HM. Antinociceptive and anti-inflammatory effects of Crocus sativus L. stigma and petal extracts in mice. BMC Pharmacol 2002; 2:7-12.

19. Hosseinzadeh H, Shariaty VM. Anti-nociceptive effect of safranal, a constituent of Crocus sativus (saffron), in mice. Pharmacologyonline 2007; 2:498-503.

20. Amin B, Hosseinzadeh H. Evaluation of aqueous and ethanolic extracts of saffron, Crocus sativus L., and its constituents, safranal and crocin in allodynia and hyperalgesia induced by chronic constriction injury model of neuropathic pain in rats. Fitoterapia 2012; 83:888-895.

21. Ochiai T, Shimeno H, Mishima K, Iwasaki K, Fujiwara M, Tanaka H, et al. Protective effects of carotenoids from saffron on neuronal injury in vitro and in vivo. Biochim Biophys Acta 2007; 1770:578-584.

22. Papandreou MA, Kanakis CD, Polissiou MG, Efthimiopoulos S, Cordopatis P, Margarity M, et al. Inhibitory activity on amyloid-beta aggregation and antioxidant properties of Crocus sativus stigmas extract and its crocin constituents. J Agric Food Chem 2006; 54:8762-8768.

23. Hosseinzadeh H, Sadeghnia HR, Rahimi A. Effect of safranal on extracellular hippocampal levels of glutamate and aspartate during kainic acid treatment in anesthetized rats. Planta Med 2008; 74:1441-1445.

24. National Research Council. Guide for the Care and Use of Laboratory Animals. Washington: National Academy Press; 1996.

25. Rahimi Shourmasti F, Goudarzi I, Lashkarbolouki T, Abrari K, Elahdadi Salmani M, Goudarzi A. Effects of riluzole on harmaline induced tremor and ataxia in rats: biochemical, histological and behavioral studies. Eur J Pharmacol 2012; 695:40-47.

26. Martin FC, Thu Le A, Handforth A. Harmaline-induced tremor as a potential preclinical screening method for essential tremor medications. Mov Disord 2005; 20: 298-305

27. Sharma PL. Mechanism of antitremorine-activity of adrenergic beta-receptor antagonists in the rat. Q J Exp Physiol Cogn Med Sci 1970; 55:202-206.

28. Biary N, Arshaduddin M, Al Deeb S, Al Moutaery K, Tariq M. Effect of lidocaine on harmaline-induced tremors in the rat. Pharmacol Biochem Behav 2000; 65:117-121.

29. Louis ED, Zheng W, Jurewicz EC, Watner D, Chen J, Factor-Litvak P, et al. Elevation of blood beta-carboline alkaloids in essential tremor. Neurology 2002; 59:1940-1944.

30. Hosseinzadeh H, Sadeghi Shakib S, Khadem Sameni A, Taghiabadi E. Acute and subacute toxicity of safranal, a constituent of saffron, in mice and rats. Iran J Pharm Res 2013; 12:93-99.

31. Hosseinzadeh H, Talebzadeh F. Anticonvulsant evaluation of safranal and crocin from Crocus sativus in mice. Fitoterapia 2005; 76:722-724.

32. Hosseinzadeh H, Khosravan V. Anticonvulsant effects of aqueous and ethanolic extracts of Crocus sativus L. stigmas in mice. Arch Im Irn Med 2002; 5:44-47.

33. Miwa H. Rodent models of tremor. Cerebellum 2007; 6:66-72.

34. Grella B, Dukat M, Young R, Teitler M, Herrick-Davis K, Gauthier CB, et al. Investigation of hallucinogenic and related beta-carbolines. Drug Alcohol Depend 1998; 50:99-107.

35. Arshaduddin M, Al Kadasah S, Biary N, Al Deeb S, Al Moutaery K, Tariq M. Citalopram, a selective serotonin reuptake inhibitor augments harmaline-induced tremor in rats. Behav Brain Res 2004; 153:15-20.

36. Georgiadou G, Tarantilis P, Pitsikas N. Effects of the active constituents of Crocus sativus L., crocins, in an animal model of obsessive compulsive disorder. Neurosci Lett 2012; 528:27-30.

37. Graf M, Kantor S, Anheuer ZE, Modos EA, Bagdy G. m-CPP-induced self-grooming is mediated by 5-HT2C receptors. Behav Brain Res 2003; 142:175-179.

38. Du W, Aloyo VJ, Harvey JA. Harmaline competitively inhibits [3H]MK-801 binding to the NMDA receptor in rabbit brain. Brain Res 1997; 770:26-29.

39. Iseri PK, Karson A, Gullu KM, Akman O, Kokturk S, Yardymoglu M, et al. The effect of memantine in harmaline-induced tremor and neurodegeneration. Neuropharmacology 2011; 61:715-723.               

40. Lechtenberg M, Schepmann D, Niehues M, Hellenbrand N, Wunsch B, Hensel A. Quality and functionality of saffron: quality control, species assortment and affinity of extract and isolated saffron compounds to NMDA and sigma1 (sigma-1) receptors. Planta Med 2008; 74:764-772.

41. Nieber K, Berger F, Hensel A. Saffron extract and trans-crocetin inhibits excitotoxicity by inhibition of postsynaptically located glutamate receptors in rat brain neurons. 6th European Congress of Pharmacology (EPHAR); 2012; Germany.

42. Tamaddonfard E, Hamzeh-Gooshchi N. Effects of intraperitoneal and intracerebroventricular injection of crocin on acute corneal pain in rats. Phytother Res 2010; 24:1463-1467.

43. Mehri S, Abnous K, Mousavi SH, Shariaty VM, Hosseinzadeh H. Neuroprotective effect of crocin on acrylamide-induced cytotoxicity in PC12 cells. Cell Mol Neurobiol 2012; 32:227-235.

44. Lari P, Abnous K, Imenshahidi M, Rashedinia M, Razavi M, Hosseinzadeh H. Evaluation of diazinon-induced hepatotoxicity and protective effects of crocin. Toxicol Ind Health 2015; 31:367-376.

45. Shinozaki H, Hirate K, Ishida M. Further studies on quantification of drug-induced tremor in mice: effects of antitremorgenic agents on tremor frequency. Exp Neurol 1985; 88:303-315.

46. Paterson NE, Malekiani SA, Foreman MM, Olivier B, Hanania T. Pharmacological characterization of harmaline-induced tremor activity in mice. Eur J Pharmacol 2009; 616:73-80.

47. Hosseinzadeh H, Sadeghnia HR. Protective effect of safranal on pentylenetetrazol-induced seizures in the rat: involvement of GABAergic and opioids systems. Phytomedicine 2007; 14:256-262.

48. Sadeghnia HR, Cortez MA, Liu D, Hosseinzadeh H, Snead OC. Antiabsence effects of safranal in acute experimental seizure models: EEG and autoradiography. J Pharm Pharm Sci 2008; 11:1-14.

49. Alavizadeh H, Hosseinzadeh H. Bioactivity assessment and toxicity of crocin: A comprehensive review. Food Chem Toxicol 2013; 64:65-80.

50. Xi L, Qian Z, Du P, Fu J. Pharmacokinetic properties of crocin (crocetin digentiobiose ester) following oral administration in rats. Phytomedicine 2007; 14:633-636.

51. Linardaki ZI, Orkoula MG, Kokkosis AG, Lamari FN, Margarity M. Investigation of the neuroprotective action of saffron (Crocus sativus L.) in aluminum-exposed adult mice through behavioral and neurobiochemical assessment. Food Chem Toxicol 2013; 52:163-170.

52. Boskabady MH, Shafei MN, Shakiba A, Sefidi HS. Effect of aqueous-ethanol extract from Crocus sativus (saffron) on guinea-pig isolated heart. Phytother 2008; 22:330-334.