• SAPTARSHI PANIGRAHI Department of Pharmacology, Key Laboratory of Neuropsychiatric Diseases, China Pharmaceutical University, Nanjing 210009, China.
  • SOMNATH SURAI Department of Pharmaceutics, Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, China.
  • HAO HONG Department of Pharmacology, Key Laboratory of Neuropsychiatric Diseases, China Pharmaceutical University, Nanjing 210009, China.


Objective: Treatment experiment was conducted to investigate the effectiveness and mechanism of the action of zileuton in corticosteroid-induced depressive mice model through neuroinflammation.

Methods: The mice were randomly separated into four groups: (Veh+Veh), (Corticosteroid+Veh), (Corticosteroid+ZIL50), and (Corticosteroid+ZIL100). Intraperitoneal injection of corticosterone (CORT) (20 mg/kg for 6 weeks) was used in the mice to induce depression and neuroinflammation diverse from the Veh+Veh group, which was injected only physiological saline. The drug-treated groups (Corticosteroid+ZIL50 and Corticosteroid+ZIL100) were orally administered with the mentioned doses of zileuton. After confirming the effectiveness of zileuton through the behavioral tests, the mechanism of the action of the drug was explored through a set of biochemical assays.

Results: Zileuton (50/100 mg/kg) administration improved the performance of the mice in the behavioral experiments (p<0.05 or 0.01). Immunohistochemistry detection of Iba1+ revealed over activation of microglial cells in the corticosteroid-treated mice which was suppressed by the zileuton (50 or 100 mg/kg [p<0.05 or 0.01]). Through Western blotting tests, it had been found that CORT (i.p.) administration led to the increment of the protein 5-Lipoxygenase in the mouse hippocampus associated with neuroinflammation, which was decreased significantly by zileuton (p<0.05 or 0.01). Level of tumor necrosis factor-alpha, interleukin-1 beta, nuclear factor kappa B p65 protein (for neuroinflammation), Bax, and cleaved caspase-3 and TUNEL assay increased, and Bcl-2 expression decreased in the CORT-induced depressive mice. These were significantly reversed by zileuton (50 or 100 mg/kg [p<0.05 or 0.01]).

Conclusion: It can be concluded that selective 5-lipoxygenase inhibitor zileuton can efficiently inhibit the depressive-like behavior/activity in CORT-induced depressive mouse model. Moreover, the underlying mechanism may be the inhibition of hippocampal neuroinflammation and apoptosis.

Keywords: Depression, Zileuton, 5-Lipoxygenase, Neuroinflammation, Apoptosis


1. Duman RS. Pathophysiology of depression and innovative treatments: Remodeling glutamatergic synaptic connections. Dialogues Clin Neurosci 2014;16:11-27.
2. Steiner MA, Marsicano G, Nestler EJ, Holsboer F, Lutz B, Wotjak CT. Antidepressant-like behavioral effects of impaired cannabinoid receptor type 1 signaling coincide with exaggerated corticosterone secretion in mice. Psychoneuroendocrinology 2008;33:54-67.
3. Brown ES, Varghese FP, McEwen BS. Association of depression with medical illness: Does cortisol play a role? Biol Psychiatry 2004;55:1-9.
4. Wulsin AC, Herman JP, Solomon MB. Mifepristone decreases depression-like behavior and modulates neuroendocrine and central hypothalamic-pituitary-adrenocortical axis responsiveness to stress. Psychoneuroendocrinology 2010;35:1100-12.
5. Sonino N, Fallo F, Fava GA. Psychosomatic aspects of Cushing’s syndrome. Rev Endocr Metab Disord 2010;11:95-104.
6. Huang Z, Zhong XM, Li ZY, Feng CR, Pan AJ, Mao QQ. Curcumin reverses corticosterone-induced depressive-like behavior and decrease in brain BDNF levels in rats. Neurosci Lett 2011;493:145-8.
7. Lee B, Shim I, Lee HJ, Yang Y, Hahm DH. Effects of acupuncture on chronic corticosterone-induced depression-like behavior and expression of neuropeptide Y in the rats. Neurosci Lett 2009;453:151-6.
8. Lan R, Xiang J, Zhang Y, Wang GH, Bao J, Li WW, et al. PI3K/Akt Pathway Contributes to Neurovascular Unit Protection of Xiao-Xu-Ming Decoction against Focal Cerebral Ischemia and Reperfusion Injury in Rats. Evid Based Complement Alternat Med 2013;2013:459467.
9. Dantzer R, O’Connor JC, Freund GG, Johnson RW, Kelley KW. From inflammation to sickness and depression: When the immune system subjugates the brain. Nat Rev Neurosci 2008;9:46-56.
10. Raison CL, Capuron L, Miller AH. Cytokines sing the blues: Inflammation and the pathogenesis of depression. Trends Immunol 2006;27:24-31.
11. Liu Y, Ho RC, Mak A. Interleukin (IL)-6, tumour necrosis factor alpha (TNF-?) and soluble interleukin-2 receptors (sIL-2R) are elevated in patients with major depressive disorder: A meta-analysis and meta-regression. J Affect Disord 2012;139:230-9.
12. Rådmark O, Werz O, Steinhilber D, Samuelsson B. 5-Lipoxygenase: Regulation of expression and enzyme activity. Trends Biochem Sci 2007;32:332-41
13. Nakagawa Y, Chiba K. Role of microglial m1/m2 polarization in relapse and remission of psychiatric disorders and diseases. Pharmaceuticals (Basel) 2014;7:1028-48.
14. Giannopoulos PF, Chu J, Joshi YB, Sperow M, Li JG, Kirby LG, et al. Gene knockout of 5-lipoxygenase rescues synaptic dysfunction and improves memory in the triple-transgenic model of Alzheimer’s disease. Mol Psychiatry 2014;19:511-8.
15. Joshi YB, Praticò D. Knockout of 5-lipoxygenase results in age-dependent anxiety-like behavior in female mice. PLoS One 2011;6:e29448.
16. Shi SS, Yang WZ, Tu XK, Wang CH, Chen CM, Chen Y. 5-Lipoxygenase inhibitor zileuton inhibits neuronal apoptosis following focal cerebral ischemia. Inflammation 2013;36:1209-17.
17. Gupta D, Radhakrishnan M, Kurhe Y. Effect of a novel 5-HT3 receptor antagonist 4i, in corticosterone-induced depression-like behavior and oxidative stress in mice. Steroids 2015;96:95-102.
18. Porsolt RD, Martin P, Lenègre A, Fromage S, Drieu K. Effects of an extract of Ginkgo Biloba (EGB 761) on “learned helplessness” and other models of stress in rodents. Pharmacol Biochem Behav 1990;36:963-71.
19. Cryan JF, Mombereau C, Vassout A. The tail suspension test as a model for assessing antidepressant activity: Review of pharmacological and genetic studies in mice. Neurosci Biobehav Rev 2005;29:571-625.
20. Baitharu I, Jain V, Deep SN, Kumar G. Exposure to hypobaric hypoxia and reoxygenation induces transient anxiety-like behavior in rat. J Behav Brain Sci 2013;3:519-602.
21. Santarelli L, Saxe M, Gross C, Surget A, Battaglia F, Dulawa S, et al. Requirement of hippocampal neurogenesis for the behavioral effects of antidepressants. Science 2003;301:805-9.
22. Graeber MB, Streit WJ. Microglia: Biology and pathology. Acta Neuropathol 2010;119:89-105.
23. Kazmi SM, Plante RK, Visconti V, Taylor GR, Zhou L, Lau CY. Suppression of NF kappa B activation and NF kappa B-dependent gene expression by tepoxalin, a dual inhibitor of cyclooxygenase and 5-lipoxygenase. J Cell Biochem 1995;57:299-310.
24. Tanuma N, Sakuma H, Sasaki A, Matsumoto Y. Chemokine expression by astrocytes plays a role in microglia/macrophage activation and subsequent neurodegeneration in secondary progressive multiple sclerosis. Acta Neuropathol 2006;112:195-204.
25. Sivakumar V, Foulds WS, Luu CD, Ling EA, Kaur C. Retinal ganglion cell death is induced by microglia derived pro-inflammatory cytokines in the hypoxic neonatal retina. J Pathol 2011;224:245-60.
26. Frakes AE, Ferraiuolo L, Haidet-Phillips AM, Schmelzer L, Braun L, Miranda CJ, et al. Microglia induce motor neuron death via the classical NF-?B pathway in amyotrophic lateral sclerosis. Neuron 2014;81:1009-23.
27. Fan H, Zhang K, Shan L, Kuang F, Chen K, Zhu K, et al. Reactive astrocytes undergo M1 microglia/macrohpages-induced necroptosis in spinal cord injury. Mol Neurodegener 2016;11:14.
28. Webster CM, Hokari M, McManus A, Tang XN, Ma H, Kacimi R, et al. Microglial P2Y12 deficiency/inhibition protects against brain ischemia. PLoS One 2013;8:e70927.
29. Zhao CZ, Zhao B, Zhang XY, Huang XQ, Shi WZ, Liu HL, et al. Cysteinyl leukotriene receptor 2 is spatiotemporally involved in neuron injury, astrocytosis and microgliosis after focal cerebral ischemia in rats. Neuroscience 2011;189:1-1.
30. Pandey DK, Yadav SK, Mahesh R, Rajkumar R. Depression-like and anxiety-like behavioural aftermaths of impact accelerated traumatic brain injury in rats: A model of comorbid depression and anxiety. Behav Brain Res 2009;205:436-42.
31. Huang XJ, Zhang WP, Li CT, Shi WZ, Fang SH, Lu YB, et al. Activation of CysLT receptors induces astrocyte proliferation and death after oxygen-glucose deprivation. Glia 2008;56:27-37.
32. Lammers CH, Schweitzer P, Facchinetti P, Arrang JM, Madamba SG, Siggins GR, et al. Arachidonate 5-lipoxygenase and its activating protein: prominent hippocampal expression and role in somatostatin signaling. J Neurochem 1996;66:147-52.
33. Raison CL, Lowry CA, Rook GA. Inflammation, sanitation, and consternation: Loss of contact with coevolved, tolerogenic microorganisms and the pathophysiology and treatment of major depression. Arch Gen Psychiatry 2010;67:1211-24.
34. Ul Ain Q, Greig NH, Nawaz MS, Rashid S, Kamal MA. Exploring N(1)-p-fluorobenzyl-cymserine as an inhibitor of 5-lipoxygenase as a candidate for Type 2 diabetes and neurodegenerative disorder treatment. CNS Neurol Disord Drug Targets 2014;13:197-202.
35. Kawano T, Matsuse H, Kondo Y, Machida I, Saeki S, Tomari S, et al. Cysteinyl leukotrienes induce nuclear factor kappa b activation and RANTES production in a murine model of asthma. J Allergy Clin Immunol 2003;112:369-74.
36. Thompson C, Cloutier A, Bossé Y, Thivierge M, Gouill CL, Larivée P, et al. CysLT1 receptor engagement induces activator protein-1- and NF-kappaB-dependent IL-8 expression. Am J Respir Cell Mol Biol 2006;35:697-704.
37. Wang XY, Tang SS, Hu M, Long Y, Li YQ, Liao MX, et al. Leukotriene D4 induces amyloid-? generation via CysLT(1)R-mediated NF-?B pathways in primary neurons. Neurochem Int 2013;62:340-7.
38. Sethi G, Sung B, Aggarwal BB. Nuclear factor-kappaB activation: From bench to bedside. Exp Biol Med (Maywood) 2008;233:21-31.
39. Ye SM, Johnson RW. Regulation of interleukin-6 gene expression in brain of aged mice by nuclear factor kappaB. J Neuroimmunol 2001;117:87-96.
40. Stadelmann C, Deckwerth TL, Srinivasan A, Bancher C, Brück W, Jellinger K, et al. Activation of caspase-3 in single neurons and autophagic granules of granulovacuolar degeneration in Alzheimer’s disease. Evidence for apoptotic cell death. Am J Pathol 1999;155:1459-66.
41. Lawrence T, Gilroy DW, Colville-Nash PR, Willoughby DA. Possible new role for NF-kappaB in the resolution of inflammation. Nat Med 2001;7:1291-7.
42. Rohn TT, Vyas V, Hernandez-Estrada T, Nichol KE, Christie LA, Head E. Lack of pathology in a triple transgenic mouse model of Alzheimer’s disease after overexpression of the anti-apoptotic protein Bcl-2. J Neurosci 2008;28:3051-9.
43. Tusi SK, Ansari N, Amini M, Amirabad AD, Shafiee A, Khodagholi F. Attenuation of NF-kappaB and activation of Nrf2 signaling by 1,2,4-triazine derivatives, protects neuron-like PC12 cells against apoptosis. Apoptosis 2010;15:738-51.
44. Lepley RA, Fitzpatrick FA. 5-Lipoxygenase compartmentalization in granulocytic cells is modulated by an internal bipartite nuclear localizing sequence and nuclear factor kappa B complex formation. Arch Biochem Biophys 1998;356:71-6.
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How to Cite
PANIGRAHI, S., S. SURAI, and H. HONG. “EFFECTS AND UNDERLYING MECHANISM OF 5-LIPOXYGENASE INHIBITOR (ZILEUTON) ON MICE DEPRESSIVE-LIKE BEHAVIOR”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 13, no. 3, Feb. 2020, pp. 180-6, doi:10.22159/ajpcr.2020.v13i3.37106.
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