Effects of troxerutin on anxiety- and depressive-like behaviors induced by chronic mild stress in adult male rats

Document Type : Original Article


1 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

2 Higher Education Institute of Rab-Rashid, Tabriz, Iran

3 Neurosciences Research Center (NSRC), Tabriz University of Medical Sciences, Tabriz, Iran


Objective(s):Chronic stress has been linked to the pathophysiology of mood disorders including anxiety and depression. In this study, we aimed to investigate the effect of troxerutin (TRX), as a flavonol, on stress-induced anxiety and depression.
Materials and Methods: 56 animals were randomly divided into seven groups (n=8 per group) as follows: control, saline, TRX 50, TRX 150, TRX 300, Diazepam, and Imipramine. Chronic mild stress (CMS) was induced by restraining animals in Plexiglas cylinders for 1 hr each day for 25 consecutive days. Different doses (50, 150, and 300 mg/kg, oral gavage) of troxerutin was gavaged for 14 consecutive days. At the end of treatments, anxiety- and depressive-like behaviors were tested using elevated plus-maze (EPM), open field test (OFT), and forced swimming test (FST).
Results: CMS significantly increased immobility (P<0.05) and decreased swimming (P<0.01) time in FST. However, different doses of troxerutin significantly decreased immobility (P<0.01) and increased swimming (P<0.001) time. CMS also significantly (P<0.01) decreased the percentage of open arm entrance (%OAE), whereas troxerutin significantly increased both %OAE and percentage of open arm time (%OAT) in the EPM. Moreover, CMS significantly decreased time spent in the center (P<0.001) and the number of center entrances (P<0.01) in the OFT. However, troxerutin significantly increased time spent in the center and number of the entrances crossing. Furthermore, CMS significantly increased serum cortisol levels and troxerutin decreased it.
Conclusion: Troxerutin demonstrated anxiolytic- and antidepressant-like activities in rodents, which supports the use of herbal medicine in the mood disorders.


Main Subjects

1. Chrousos GP. Stress and disorders of the stress system. Nat Rev Endocrinol 2009; 5:374-381.
2. Steel Z, Marnane C, Iranpour C, Chey T, Jackson JW, Patel V, et al. The global prevalence of common mental disorders: a systematic review and meta-analysis 1980 – 2013. Int J Epidemiol 2014; 43:476-493.
3. Bechtold AG, Patel G, Hochhaus G, Scheuer DA. Chronic blockade of hindbrain glucocorticoid receptors reduces blood pressure responses to novel stress and attenuates adaptation to repeated stress. Am J Physiol Regul Integr Comp 2009; 296:1445-1454.
4.  Xie X, Chi X, Zhou W, Ma Y, Zhang Y. Progress in research of animal stress models. Chinese Journal of New Drugs 2008; 17:1375-1380.
5. Varlinskaya EI, Doremus-Fitzwater TL, Spear LP. Repeated restraint stress alters sensitivity to the social consequences of ethanol in adolescent and adult rats. Pharmacol Biochem Behav 2010; 96:228-235.
6. Sevgi S, Ozek M, Eroglu L. L-NAME prevents anxiety-like and depression-like behavior in rats exposed to restraint stress. Methods Find Exp Clin Pharmacol 2006; 28:95-100.
7. Chotiwat C, Harris RB. Increased anxiety-like behavior during the post-stress period in mice exposed to repeated restraint stress. Horm Behav 2006; 50:489-495.
8. Herman JP, McKlveen JM, Ghosal S, Kopp B, Wulsin A, Makinson R, et al. Regulation of the hypothalamic-pituitary-adrenocortical stress response. Compr Physiol 2016; 6:603-621.
9. Stetler C, Miller GE. Depression and hypothalamic-pituitary-adrenal activation: a quantitative summary of four decades of research. Psychosom Med 2011; 73:114-126.
10. Bauer ME, Perks P, Lightman SL, Shanks N. Restraint stress is associated with changes in glucocorticoid immunoregulation. Physiol Behav 2001; 73:525-532.
11. Ottenweller JE, Servatius RJ, Tapp WN, Drastal SD, Bergen MT, Natelson BH. A chronic stress state in rats: effects of repeated stress on basal corticosterone and behavior. Physiol Behav 1992; 51:689-698.
12. Heidarzadeh F, Badalzadeh R, Hatami H. The effect of troxerutin on lipid peroxidation and tissue injury induced by myocardial ischemia reperfusion injury in diabetic rat. Razi J Med Sci 2014; 21:37-45.
13. Riccioni C, Sarcinella R, Izzo A, Palermo G, Liguori M. Effectiveness of Troxerutin in association with Pycnogenol in the pharmacological treatment of venous insufficiency. Minerva cardioangiologica 2004; 52:43-48.
14. Fan SH, Zhang ZF, Zheng YL, Lu J, Wu DM, Shan Q, et al. Troxerutin protects the mouse kidney from d-galactose-caused injury through anti-inflammation and anti-oxidation. Int Immunopharmacol 2009; 9:91-96.
15. Yang X, Wang F, Hu S. The electrochemical oxidation of troxerutin and its sensitive determination in pharmaceutical dosage forms at PVP modified carbon paste electrode. Colloids and Surfaces B: Biointerfaces 2006; 52:8-13.
16. Maurya DK, Balakrishnan S, Salvi VP, Nair CKK. Protection of cellular DNA from γ-radiation-induced damages and enhancement in DNA repair by troxerutin. Mol Cell Biochem 2005; 280:57-68.
17. Farajdokht F, Amani M, Bavil FM, Alihemmati A, Mohaddes G, Babri S. Troxerutin protects hippocampal neurons against amyloid beta-induced oxidative stress and apoptosis. EXCLI J 2017; 16:1081-1089.
18. Lu J, Wu DM, Zheng YL, Hu B, Cheng W, Zhang ZF, et al. Troxerutin counteracts domoic acid–induced memory deficits in mice by inhibiting CCAAT/enhancer binding protein β–mediated inflammatory response and oxidative stress. J Immunol 2013; 190:3466-3479.
19. Zhang ZF, Fan SH, Zheng YL, Lu J, Wu DM, Shan Q, et al. Troxerutin protects the mouse liver against oxidative stress-mediated injury induced by D-galactose. J Agric Food Chem 2009; 57:7731-7736.
20. Lu J, Wu DM, Zheng ZH, Zheng YL, Hu B, Zhang ZF. Troxerutin protects against high cholesterol-induced cognitive deficits in mice. Brain 2011; 134:783-797.
21. Lu J, Wu DM, Hu B, Cheng W, Zheng YL, Zhang ZF, et al. Chronic administration of troxerutin protects mouse brain against D-galactose-induced impairment of cholinergic system. Neurobiol Learn Mem 2010; 93:157-164.
22. Zhang LM, Yao JZ, Li Y, Li K, Chen HX, Zhang YZ, et al. Anxiolytic effects of flavonoids in animal models of posttraumatic stress disorder. Evid Based Complement Alternat Med 2012; 2012:623753.
23. An L, Zhang YZ, Yu NJ, Liu XM, Zhao N, Yuan L, et al. Role for serotonin in the antidepressant-like effect of a flavonoid extract of Xiaobuxin-Tang. Pharmacol Biochem Behav 2008; 89:572-580.
24. An L, Zhang YZ, Yu NJ, Liu XM, Zhao N, Yuan L, et al. The total flavonoids extracted from Xiaobuxin-Tang up-regulate the decreased hippocampal neurogenesis and neurotrophic molecules expression in chronically stressed rats. Prog Neuropsychopharmacol Biol Psychiatry 2008; 32:1484-1490.
25. An L, Zhang YZ, Liu XM, Yu NJ, Chen HX, Zhao N, et al. Total flavonoids extracted from xiaobuxin-tang on the hyperactivity of hypothalamic-pituitary-adrenal axis in chronically stressed rats. J Evid Based Complementary Altern Med 2011; 2011:367619.
26. Hendriksen H, Prins J, Olivier B, Oosting RS. Environmental enrichment induces behavioral recovery and enhanced hippocampal cell proliferation in an antidepressant-resistant animal model for PTSD. PLoS One 2010; 5:e11943.
27. Farajdokht F, Soleimani M, Mehrpouya S, Barati M, Nahavandi A. The role of hepcidin in chronic mild stress-induced depression. Neurosci Lett 2015; 588:120-124.
28. Farajdokht F, Babri S, Karimi P, Alipour MR, Bughchechi R, Mohaddes G. Chronic ghrelin treatment reduced photophobia and anxiety‐like behaviors in nitroglycerin‐induced migraine: role of pituitary adenylate cyclase‐activating polypeptide. Eur J Neurosci 2017; 45:763-772.
29. Shahsavary F, Abbasnejad M. Effect of co-administration of ascorbic acid and bromocriptine in nucleus accumbens shell on locomotor activity in male rats by Open Field test. Physiol Pharmacol 2013; 17:125-136.
30. Walf AA, Frye CA. The use of the elevated plus maze as an assay of anxiety-related behavior in rodents. Nat Protoc 2007; 2:322-328.
31. Sajdyk TJ, Fitz SD, Shekhar A. The role of neuropeptide Y in the amygdala on corticotropin-releasing factor receptor-mediated behavioral stress responses in the rat. Stress 2006; 9:21-28.
32. Waters P, McCormick CM. Caveats of chronic exogenous corticosterone treatments in adolescent rats and effects on anxiety-like and depressive behavior and hypothalamic-pituitary-adrenal (HPA) axis function. Biol Mood Anxiety Disord 2011; 1:4.
33. Zhu S, Shi R, Wang J, Wang JF, Li XM. Unpredictable chronic mild stress not chronic restraint stress induces depressive behaviours in mice. Neuroreport 2014; 25:1151-1155.
34. Slattery DA, Cryan JF. Using the rat forced swim test to assess antidepressant-like activity in rodents. Nat Protoc 2012; 7:1009-10014.
35. Cryan JF, Holmes A. The ascent of mouse: advances in modelling human depression and anxiety. Nat Rev Drug Discov 2005; 4:775-790.
36. Gregus A, Wintink AJ, Davis AC, Kalynchuk LE. Effect of repeated corticosterone injections and restraint stress on anxiety and depression-like behavior in male rats. Behav Brain Res 2005; 156:105-114.
37. Slavich GM, Irwin MR. From Stress to Inflammation and Major Depressive Disorder: A Social Signal Transduction Theory of Depression. Psychol Bull 2014; 140:774-815.
38. Köhler O, Benros ME, Nordentoft M, Farkouh ME, Iyengar RL, Mors O, et al. Effect of anti-inflammatory treatment on depression, depressive symptoms, and adverse effects: A systematic review and meta-analysis of randomized clinical trials. JAMA Psychiatry 2014; 71:1381-1391.
39. Panat NA, Maurya DK, Ghaskadbi SS, Sandur SK. Troxerutin, a plant flavonoid, protects cells against oxidative stress-induced cell death through radical scavenging mechanism. Food Chem 2016; 194:32-45.
40. Lee KS, Cha HJ, Lee GT, Lee KK, Hong JT, Ahn KJ, et al. Troxerutin induces protective effects against ultraviolet B radiation through the alteration of microRNA expression in human HaCaT keratinocyte cells. Int J Mol Med 2014; 33:934-942.
41. Babri S, Amani M, Mohaddes G, Alihemmati A, Ebrahimi H. Protective effects of troxerutin on β-amyloid (1-42)-induced impairments of spatial learning and memory in rats. Neurophysiol 2012: 44:387-393.
42. Babri S, Mohaddes G, Feizi I, Mohammadnia A, Niapour A, Alihemmati A, et al. Effect of troxerutin on synaptic plasticity of hippocampal dentate gyrus neurons in a β-amyloid model of Alzheimer,s disease: An electrophysiological study. Eur J Pharmacol 2014; 732:19-25.
43. Goshen I, Kreisel T, Licht T, Weidenfeld J, Ben-Hur T, Yirmiya R. Brain interleukin-1 mediates chronic stress-induced depression in mice via adrenocortical activation and hippocampal neurogenesis suppression. ‎Mol Psychiatry 2008; 13:717-728.
44. Dal-Zotto S, Martı́ O, Armario A. Influence of single or repeated experience of rats with forced swimming on behavioural and physiological responses to the stressor. Behav Brain Res 2000; 114:175-181.
45. Kusnecov AW, Rabin BS. Stressor-lnduced Alterations of Immune Function: Mechanisms and Issues. Int Arch Allergy Immunol 1994; 105:107-121.
46. Hargreaves KM. Neuroendocrine markers of stress. Anesth Prog 1990; 37:99–105.
47. Juster RP, McEwen BS, Lupien SJ. Allostatic load biomarkers of chronic stress and impact on health and cognition. Neurosci Biobehav Rev 2010; 35:2-16.
48. Bhatnagar S, Dallman M. Neuroanatomical basis for facilitation of hypothalamic-pituitary-adrenal responses to a novel stressor after chronic stress. Neuroscience 1998; 84:1025-1039.
49. Parker KJ, Schatzberg AF, Lyons DM. Neuroendocrine aspects of hypercortisolism in major depression. Horm Behav 2003; 43:60-66.
50. Burke HM, Davis MC, Otte C, Mohr DC. Depression and cortisol responses to psychological stress: a meta-analysis. Psychoneuroendocrinol 2005; 30:846-856.
51. Burke HM, Fernald LC, Gertler PJ, Adler NE. Depressive symptoms are associated with blunted cortisol stress responses in very low-income women. Psychosom Med 2005; 67:211-216.
52. Young EA, Altemus M, Lopez JF, Kocsis JH, Schatzberg AF, Zubieta JK. HPA axis activation in major depression and response to fluoxetine: a pilot study. Psychoneuroendocrinol 2004; 29:1198-1204.
53. Reul J, Stec I, Söder M, Holsboer F. Chronic treatment of rats with the antidepressant amitriptyline attenuates the activity of the hypothalamic-pituitary-adrenocortical system. Endocrinol 1993; 133:312-320.
54. Lee RS, Tamashiro KL, Yang X, Purcell RH, Harvey A, Willour VL, et al. Chronic corticosterone exposure increases expression and decreases deoxyribonucleic acid methylation of Fkbp5 in mice. Endocrinol 2010; 151:4332-4343.