• 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: The experiment aimed to find out the effectiveness of Zileuton, a 5-LOX inhibitor on depressive behavior and neuroinflammation in vivo.

Method: Male ICR mice (25-30g) randomly distributed Veh+Veh, CRS+Vehicle, CRS+ZIL50, and CRS+ZIL100. Zileuton was orally given in the treatment groups for 21 days after 3 weeks of stress induce CRS model. Starting from the day 1, in CRS model, mice were immobilized 8 hr/day for consecutive 21 days to induce stress. After completing the drug administration, subjected the mice for behavioral tests, and then performed histopathological & Western Blotting.

Result: Stress induces CRS model guide to the significant depressive-like behavior of the mice in behavioral tests which was united by adverse changes at the cellular/molecular level responsible for regulation of inflammatory and apoptotic processes. CRS triggered Microglial over activation in the DG of the hippocampus, which was successfully inhibited by Zileuton post-treatment at the dose of 100mg/kg than 50mg/kg. Level of TNF-α, IL- 1β, nuclear NF-κB p65, Bax, and cleaved Caspase-3 was high and Bcl-2 expression was low in the stress induce CRS -treated mice which were found to be opposite in the Zileuton (100mg/kg). However, the dose of 50mg/kg less to mimic the effects as exhibited more by the 100mg/kg dose of Zileuton.

Conclusion: It can be concluded that selective 5-lipoxygenase inhibitor Zileuton can efficiently inhibit the depressive-like behavior/activity in CRS-induced depressive mouse model. The study is the first to show the role of 5-lipoxygenase enzyme in and Chronic Restraint Stress (CRS)-induced mice models of stress, anxiety or depression.

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


1. Harvey BH, Brand L, Jeeva Z, Stein DJ. Cortical/hippocampal monoamines, HPA-axis changes and aversive behavior following stress and restress in an animal model of post-traumatic stress disorder. Physiol Behav 2006;87:881-90.
2. Zhang Y, Liu W, Zhou Y, Ma C, Li S, Cong B. Endoplasmic reticulum stress is involved in restraint stress-induced hippocampal apoptosis and cognitive impairments in rats. Physiol Behav 2014;131:41-8.
3. Ferraz AC, Delattre AM, Almendra RG, Sonagli M, Borges C, Araujo P, et al. Chronic ?-3 fatty acids supplementation promotes beneficial effects on anxiety, cognitive and depressive-like behaviors in rats subjected to a restraint stress protocol. Behav Brain Res 2011;219:116-22.
4. Kim KS, Han PL. Optimization of chronic stress paradigms using anxiety-and depression-like behavioral parameters. J Neurosci Res 2006;83:497-507.
5. Chaudhari N, Talwar P, Parimisetty A, Lefebvre d’Hellencourt C, Ravanan P. A molecular web: Endoplasmic reticulum stress, inflammation, and oxidative stress. Front Cell Neurosci 2014;8:213.
6. 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.
7. 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.
8. Rådmark O, Werz O, Steinhilber D, Samuelsson B. 5-Lipoxygenase: Regulation of expression and enzyme activity. Trends Biochem Sci 2007;32:332-41.
9. Nakagawa Y, Chiba K. Role of microglial M1/M2 polarization in relapse and remission of psychiatric disorders and diseases. MDPI Pharm 2014;7:1028-48.
10. 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.
11. Joshi YB, Praticò D. Knockout of 5-lipoxygenase results in age-dependent anxiety-like behavior in female mice. PLoS One 2011;6:e29448.
12. Chiba S, Numakawa T, Ninomiya M, Richards MC, Wakabayashi C, Kunugi H. Chronic restraint stress causes anxiety- and depression-like behaviors, downregulates glucocorticoid receptor expression, and attenuates glutamate release induced by brain-derived neurotrophic factor in the prefrontal cortex. Prog Neuropsychopharmacol Biol Psychiatry 2012;39:112-9.
13. 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.
14. 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.
15. Baitharu I, Jain V, Deep SN, Kumar G, Ilavazhagan G. Exposure to hypobaric hypoxia and reoxygenation induces transient anxiety-like behavior in rat. J Behav Brain Sci 2013;3:519-602.
16. 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.
17. Graeber MB, Streit WJ. Microglia: Biology and pathology. Acta Neuropathol 2010;119:89-105.
18. 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.
19. 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.
20. 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.
21. 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.
22. 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.
23. 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.
24. 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.
25. 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.
26. 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.
27. Sethi G, Sung B, Aggarwal BB. Nuclear factor-kappaB activation: From bench to bedside. Exp Biol Med (Maywood) 2008;233:21-31.
28. 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.
29. 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.
30. 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.
31. 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.
32. 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.
33. 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.
135 Views | 40 Downloads
How to Cite
PANIGRAHI, S., S. SURAI, and H. HONG. “5-LIPOXYGENASE INHIBITOR (ZILEUTON) PRODUCES ANTI-DEPRESSION EFFECT IN STRESS INDUCE CRS MICE MODEL”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 13, no. 2, Dec. 2019, pp. 154-62, doi:10.22159/ajpcr.2020.v13i2.36501.
Original Article(s)