Involvement of brain-derived neurotrophic factor (BDNF) on malathion induced depressive-like behavior in subacute exposure and protective effects of crocin

Document Type : Original Article


1 School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

2 Pharmaceutical Research Center, Department of Pharmacodynamy and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

3 Pharmaceutical Research Center, Department of Medicinal Chemistry, Mashhad University of Medical Sciences, Mashhad, Iran

4 Targeted Drug Delivery Research Center, Department of Pharmacodynamy and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran


Objective(s): In this study the effect of crocin, a carotenoid isolated from saffron, on malathion (an organophosphate insecticide) induced depressive- like behavior in subacute exposure was investigated. Moreover the molecular mechanism of malathion induced depressive- like behavior and its decreasing effect on the level of brain derived neurotrophic factor (BDNF) in rat hippocampus and cerebral cortex were evaluated.
Materials and Methods: Male Wistar rats were exposed to malathion (50 mg/kg/day, IP) alone or in combination with crocin (10, 20 and 40 mg/kg/day, IP), imipramine (20 mg/kg/day, IP) and vitamin E (200 mg/kg, three times a week, IP) respectively for 14 days. The forced swimming test (FST) was performed on days 1st, 7th and 14st. The level of malondealdehyde (MDA) and reduced glutathione (GSH) were measured in cerebral cortex and hippocampus of rats. The protein level of BDNF was evaluated using Western blot analysis.
Results: Malathion (50 mg/kg, IP) increased immobility time in the FST, without affecting total locomotor activity in open-field test. Malathion increased the malondealdehyde (MDA) and decreased the glutathione (GSH), whereas these effects were reversed by crocin and vitamin E. Malathion decreased plasma acetylcholinesterase  activity,  however  this effect was not reversed by crocin or vitamin E. Malathion reduced the protein level of BDNF in rat hippocampus. Imipramine and crocin  prevented the decreasing effect of malathion on BDNF.
Conclusion: These results showed that crocin attenuates some neurochemical and behavioral effects induced by malathion. This neuroprotective effect of crocin may be in part due to its effect on BDNF.


1. Delgado EHB, Streck EL, Quevedo JL, Dal-Pizzol F. Mitochondrial respiratory dysfunction and oxidative stress after chronic malathion exposure. Neurochem Res 2006; 31:1021-1025.
2. Salvi RM, Lara DR, Ghisolfi ES, Portela LV, Dias RD, Souza DO. Neuropsychiatric evaluation in subjects chronically exposed to organophosphate pesticides. Toxicol Sci  2003; 72:267-271.
3. Haenisch B, Bönisch H. Depression and antidepressants. Insights from knockout of dopamine, serotonin or noradrenaline re-uptake transporters. Pharmacol Ther 2011; 129:352-368.
4. Hisaoka K, Maeda N, Tsuchioka M, Takebayashi M. Antidepressants induce acute CREB phosphorylation and CRE-mediated gene expression in glial cells: a possible contribution to BDNF production. Brain Res 2008; 1196:53-58.
5. Aydemir O, Devecia A, Tanelib F. The effect of chronic antidepressant treatment on serum brain-derived neurotrophic factor levels in depressed patients: a preliminary study. Prog Neuropsychopharmacol Biol Psychiatry 2005; 29:261-265.
6. Soeda S, Ochiai T, Shimeno H, Saito H, Abe K, Tanaka H, Shoyama Y. Pharmacollogical activities of crocin in saffron. J Nat Med 2007; 61:102-111.
7. Fernández J. Anticancer properties of saffron, Crocus sativus Linn. Adv Phytomed  2006; 313-30.
8. Assimopoulou A, Sinakos Z, Papageorgiou V. Radical scavenging activity of Crocus sativus L. extract and its bioactive constituents. Phytother Res 2005; 19:997-1000.
9. 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 2010; 5:419-424.
10. Sheng L, Qian Z, Zheng S, Xi L. Mechanism of hypolipidemic effect of crocin in rats: crocin inhibits pancreatic lipase. Eur J Pharmacol 2006; 543:116-122.
11. Hosseinzadeh H, Younesi MH. Antinociceptive and anti-inflammatory effects of Crocus sativus L. stigma and petal extracts in mice. BMC Pharmacol  2002;  2:7.
12. Hosseinzadeh H, Khosravan V. Anticonvulsant effects of aqueous and ethanolic extracts of Crocus sativus L. stigmas in mice. Arch Iran Med 2002; 44-47.
13. Abe K, Saito H.  Effects of saffron extract and its constituent crocin on learning behaviour and long-term potentiation. Phytother Res 2000; 14:149-52.
14. 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.
15. Hariri A, Moallem S, Mahmoudi M, Memar B, Hosseinzadeh H. Sub-acute effects of diazinon on biochemical indices and specific biomarkers in rats: Protective effects of crocin and safranal. Food Chem Toxicol 2010; 48:2803-8.
16. Mehri S, Abnous K, Mousavi S, Motamed Shariaty V, Hosseinzadeh H. Neuroprotective effect of crocin on acrylamide-induced cytotoxicity in PC12 cells. Cell Mol Neurobiol 2012; 32:227-35.
17. Razavi B, Hosseinzadeh H, Movassaghi A, Imenshahidi M, Abnous K. Protective effect of crocin on diazinon induced cardiotoxicity in subcronic exposure. Chem Boil Inter 2013; 25:547-55.
18. Akhondzadeh S, Fallah-Pour  H, Afkham K, Jamshidi A, Khalighi-Cigaroudi F. Comparison of Crocus sativus L. and imipramine in the treatment of mild to moderate depression: A pilot double-blind randomized trial. BMC Complem Altern Med  2004; 4:12-17.
19. Vahdati Hassani F, Naseri V, Razavi BM, Mehri S, Abnous K, Hosseinzadeh H. Antidepressant effects of crocin and its effects on transcript and protein levels of CREB, BDNF, and VGF in rat hippocampus. Daru 2014; 8:22.
20. Sanchez-Mateo C, Bonkanka C, Prado  B, Rabanal R. Antidepressant activity of some Hypericum reflexum L. fil. extracts in the forced swimming test in mice. J Ethnopharmacol 2007; 112:115-121.
21. Hadizadeh F, Mohajeri S, Saifi M. Extraction and purification of crocin from saffron stigmas employing a simple and efficient crystallization method. Pak J Biol Sci 2010; 13:691-698.
22. Porsolt D, Le P, Jalfre M. Depression: a new animal model sensitive to antidepressant treatments. Nature 1977; 266:730.
23. Pardon M. Perez Diaz F, Joubert C, Cohen Salmon C. Age dependent effects of a chronic ultramild stress procedure on open field behavior in B6D2F1 female mice. Physiol Behavio 2000; 70:7-13.
24. Fernandez J, Perez-Alvarez J, Fernandez-lopez J. Thiobarbituric acid test for monitoring lipid oxidation in meat. Food Chem Toxicol 1997; 99:345-53.
25. Moron M, Depierre J, Mannervik B. Levels of glutathione, glutathione reductase and glutathione S transferase activities in rat lung and liver. Biochim Biophys Acta 1979; 582:67-78.
26. Rezg R, Mornagui B, El-Fazaa S, Gharbi N. Biochemical evaluation of hepatic damage in subchronic exposure to malathion in rats: effect on superoxide dismutase and catalase activities using native PAGE. C R Biol 2008; 331:655-62.
27. Chen N, Luo D, Yao X, Yu C, Wang Y, Wang Q. Pesticides Induce Spatial Memory Deficits with Synaptic Impairments and an Imbalanced Tau Phosphorylation in Rats. J Alzheimers Dis 2012; 30:585–594.
28. Brocardo P, Assini F, Franco J, Pandolfo P, Muller M, Takahashi RN, et al. Zinc attenuates Malathion-Induced Depressant-like Behavior and Confers Neuroprotection in the Rat Brain. Toxicology 2007; 97:140-8.
29. Ramos ZR, Fortunato JJ, Agostinho FR, Martins MR, Correa M, Schetinger MR, et al. Influence of malathion on acetylcholinesterase activity in rats submitted to a forced swimming test. Neurotox Res  2006; 9:285-290.
30. Walsh RN, Cummins RA. The open-field test: A critical review. Psychol Bull 1976; 83: 482–504.
31. Assini FL, Zanette KD, Brocardo PS, Pandolfo P, Rodrigues ALS, Takahashi RN. Behavioral effects and ChE measures after acute and repeated administration of malathion. Environ Toxicol Pharmacol 2005; 20:443–449.
32. Alavizadeh S, Hosseinzadeh H.  Bioactivity assessment and toxicity of crocin: a Comprehensive Review. Food Chem Toxicol 2014; 64:65-80.
33. Hosseinzadeh H, Karimi G, Niapoor M. Anti-depressant effects of Crocus sativus stigma extracts and its constituents, crocin and safranal, in mice. J Med Plants 2004; 3:48-58.
34. Nakamura K, Tanaka Y. Antidepressant-like effects of aniracetam in aged ratsand its mode of action. Psychopharmacol J 2001; 158: 205-212.
35. Akhgari M, Abdollahi M, Kebryaeezadeh A, Hosseini R, Sazevari O. Biochemical evidence for free radical-induced lipid peroxidation as a mechanism for subchronic toxicity of malathion in blood and liver of rats. Hum Exp Toxicol 2003; 22:205-211.
36. Possamai FP, Fortunato JJ, Feier G, Agostinho FR, Quevedo J, Filho DW, et al.  Oxidative stress after acute and sub-chronic malathion intoxication in Wistar rats. Env Toxicol Pharmacol 2007; 23:198-204.
37. Fortunato J J, Feier G, Vitali A, Petronilho F, Dal-Pizzol F, Quevedo J. Malathion-induced Oxidative Stress in Rat Brain Regions. Neurochem Res 2006; 31:671-678.
38.  Ghazi-Khansari M, Baeeri M, Abdollahi M. Protection by pentoxifylline of malathion-induced toxic stress and mitochondrial damage in rat brain. Hum Exp Toxicol 2010; 29:851-864.
39.  Abdollahi M,  Mostafalou S,  Pournourmohammadi SH, Shadnia SH. Oxidative stress and cholinesterase inhibition in saliva and plasma of rats following subchronic exposure to malathion. Comp Biochem Physiol Part C: Toxicol Pharmacol 2004; 137:29–34.
40. Ogutcu A, Uzunhisarcikli M, Kalender S, Durak D, Bayrakdar F, Kalender Y. The effects of organophosphate insecticide diazinon on malondialdehyde levels and myocardial cells in rat heart tissue and protective role of vitamin E. Pest Biochemi Physiol 2006; 86:93-98.
41. John S, Kale M, Rathore N, Bhatnagar D. Protective effect of Vitamin E in dimethoate and malathion induced oxidative stress in rat erythrocytes. J Nutr Biochem  2001; 12:500-504.
42. Razavi BM, Hosseinzadeh H, Abnous K, Imenshahidi M. Protective effect of crocin on diazinon induced vascular toxicity in subchronic exposure in rat aorta ex-vivo. Drug Chem Toxicol Early Online. 2014; 1–6.
43. 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 [Epub ahead of print]. 2013.
44. Wang J, Dranovsky A, Hen R. The When and Where of BDNF and the Antidepressant Response. Biol Psychiatry 2008; 63:640-641.
45. Thakker-Varia S, Alder J. Neuropeptides in depression: role of VGF. Behav Brain Res 2009; 197: 262–278.