Anticonvulsant and ameliorative effects of pioglitazone on cognitive deficits, inflammation and apoptosis in the hippocampus of rat pups exposed to febrile seizure

Document Type: Original Article


1 Rayan Center for Neuroscience & Behavior, Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Iran

2 Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran


Objective(s): Pioglitazone (PGZ), a peroxisome proliferator-activated receptor gamma (PPAR-γ) agonist, has significant neuroprotective effects and has been reported to regulate inflammatory processes.
Materials and Methods: We evaluated the effects of PGZ on febrile seizure (FS) in rat pups. Three groups of male rat pups received intraperitoneal (IP) injections of PGZ (5, 10, and 20 mg/kg). Lipopolysaccharide (LPS) and kainic acid (KA) were injected to induce FS. The rat pups behaviors were recorded and analyzed. Seizure latency, duration, and severity were recorded to evaluate the effect of PGZ on FS. Novel object recognition task (NORT) was used to evaluate the effect of PGZ on cognitive deficits induced by FS. At the end of the experimental protocol, molecular and histological tests were done.
Results: PGZ significantly increased seizure latency and decreased seizure duration and median of seizure scores (P<0.05, P<0.01, and P<0.001) after induction of FS. Rat pups exposed to FS had memory deficits both in short-term and long-term memories in the NORT that were reversed by PGZ-treatment (P<0.01 and P<0.001). PGZ significantly reduced interleukin-1β, tumor necrosis factor-α, and inducible nitric oxide synthase concentration in the hippocampus (P<0.05 and P<0.01). In addition, PGZ decreased the number of degenerating and TUNEL positive neurons in CA1, CA3, and DG subfields of the hippocampus (P< 0.05, P<0.01 and P<0.001).
Conclusion: The present results indicated that PGZ had anticonvulsant, anti-inflammatory, and anti-apoptotic effects with ameliorative effects on cognitive deficits induced by FS in rat pups.


Main Subjects

1. van Gassen KL, Hessel EV, Ramakers GM, Notenboom RG, Wolterink-Donselaar IG, Brakkee JH, et al. Characterization of febrile seizures and febrile seizure susceptibility in mouse inbred strains. Genes Brain Behav 2008; 7:578-586.
2. Feng B, Chen Z. Generation of febrile seizures and subsequent epileptogenesis. Neurosci Bull 2016; 32:481-492.
3. Eun BL, Abraham J, Mlsna L, Kim MJ, Koh S. Lipopolysaccharide potentiates hyperthermia-induced seizures. Brain Behav 2015; 5:e00348.
4. Dey A, Kang X, Qiu J, Du Y, Jiang J. Anti-inflammatory small molecules to treat seizures and epilepsy: from bench to bedside. Trends Pharmacol Sci 2016; 37:463-484.
5. Yu N, Liu H, Di Q. Modulation of Immunity and the Inflammatory Response: A New Target for Treating Drug-resistant Epilepsy. Curr Neuropharmacol 2013; 11:114-127.
6. Choy M, Dube CM, Ehrengruber M, Baram TZ. Inflammatory processes, febrile seizures, and subsequent epileptogenesis. Epilepsy Curr 2014; 14:15-22.
7. Alheim K, Bartfai T. Interleukin‐1 System: Receptors, Ligands, and ICE in the Brain and Their Involvement in the Fever Response. Ann N Y Acad Sci 1998; 840:51-58.
8. Cartmell T, Luheshi GN, Rothwell NJ. Brain sites of action of endogenous interleukin-1 in the febrile response to localized inflammation in the rat. J Physiol 1999; 518 ( Pt 2):585-594.
9. Rao RS, Prakash A, Medhi B. Role of different cytokines and seizure susceptibility: a new dimension towards epilepsy research. Indian J Exp Biol 2009; 47:625-634.
10. Viviani B, Bartesaghi S, Gardoni F, Vezzani A, Behrens M, Bartfai T, et al. Interleukin-1β enhances NMDA receptor-mediated intracellular calcium increase through activation of the Src family of kinases. J Neurosci 2003; 23:8692-8700.
11. Patel DC, Wallis G, Dahle EJ, McElroy PB, Thomson KE, Tesi RJ, et al. Hippocampal TNFalpha signaling contributes to seizure generation in an infection-induced mouse model of limbic epilepsy. eNeuro 2017; 4.
12. Mollace V, Muscoli C, Masini E, Cuzzocrea S, Salvemini D. Modulation of prostaglandin biosynthesis by nitric oxide and nitric oxide donors. Pharmacol Rev 2005; 57:217-252.
13. Brown GC, Bal-Price A. Inflammatory neurodegeneration mediated by nitric oxide, glutamate, and mitochondria. Mol Neurobiol 2003; 27:325-355.
14. Lind M, Hayes A, Caprnda M, Petrovic D, Rodrigo L, Kruzliak P, et al. Inducible nitric oxide synthase: Good or bad? Biomed Pharmacother 2017; 93:370-375.
15. Chang YC, Kuo YM, Huang AM, Huang CC. Repetitive febrile seizures in rat pups cause long-lasting deficits in synaptic plasticity and NR2A tyrosine phosphorylation. Neurobiol Dis 2005; 18:466-475.
16. Heida JG, Boisse L, Pittman QJ. Lipopolysaccharide-induced febrile convulsions in the rat: short-term sequelae. Epilepsia 2004; 45:1317-1329.
17. Heida JG, Teskey GC, Pittman QJ. Febrile Convulsions Induced by the Combination of Lipopolysaccharide and Low‐dose Kainic Acid Enhance Seizure Susceptibility, Not Epileptogenesis, in Rats. Epilepsia 2005; 46:1898-1905.
18. Eriksson C, Tehranian R, Iverfeldt K, Winblad B, Schultzberg M. Increased expression of mRNA encoding interleukin-1beta and caspase-1, and the secreted isoform of interleukin-1 receptor antagonist in the rat brain following systemic kainic acid administration. J Neurosci Res 2000; 60:266-279.
19. Rosen ED, Spiegelman BM. PPARγ: a nuclear regulator of metabolism, differentiation, and cell growth. J Biol Chem 2001; 276:37731-37734.
20. Cariou B, Charbonnel B, Staels B. Thiazolidinediones and PPARγ agonists: time for a reassessment. Trends Endocrinol Metab 2012; 23:205-215.
21. Xing B, Xin T, Hunter RL, Bing G. Pioglitazone inhibition of lipopolysaccharide-induced nitric oxide synthase is associated with altered activity of p38 MAP kinase and PI3K/Akt. J Neuroinflammation 2008; 5:4.
22. Shafaroodi H, Hassanipour M, Mousavi Z, Rahimi N, Dehpour AR. The Effects of Sub-Chronic Treatment with Pioglitazone on the Septic Mice Mortality in the Model of Cecal Ligation and Puncture: Involvement of Nitric Oxide Pathway. Acta Med Iran 2015; 53:608-616.
23. Shafaroodi H, Moezi L, Ghorbani H, Zaeri M, Hassanpour S, Hassanipour M, et al. Sub-chronic treatment with pioglitazone exerts anti-convulsant effects in pentylenetetrazole-induced seizures of mice: The role of nitric oxide. Brain Res Bull 2012; 87:544-550.
24. Abdallah DM. Anticonvulsant potential of the peroxisome proliferator-activated receptor gamma agonist pioglitazone in pentylenetetrazole-induced acute seizures and kindling in mice. Brain Res 2010; 1351:246-253.
25. Heida JG, Pittman QJ. Causal links between brain cytokines and experimental febrile convulsions in the rat. Epilepsia 2005; 46:1906-1913.
26. Drexel M, Preidt AP, Sperk G. Sequel of spontaneous seizures after kainic acid-induced status epilepticus and associated neuropathological changes in the subiculum and entorhinal cortex. Neuropharmacology 2012; 63:806-817.
27. Rezvani ME, Roohbakhsh A, Mosaddegh MH, Esmailidehaj M, Khaloobagheri F, Esmaeili H. Anticonvulsant and depressant effects of aqueous extracts of Carum copticum seeds in male rats. Epilepsy Behav 2011; 22:220-225.
28. Zhou J, Wang F, Zhang J, Gao H, Yang Y, Fu R. Repeated febrile convulsions impair hippocampal neurons and cause synaptic damage in immature rats: neuroprotective effect of fructose-1,6-diphosphate. Neural Regen Res 2014; 9:937-942.
29. Ghadimkhani M, Saboory E, Roshan-Milani S, Mohammdi S, Rasmi Y. Effect of magnesium sulfate on hyperthermia and pentylen-tetrazol-induced seizure in developing rats. Iran J Basic Med Sci 2016; 19:608-614.
30. Reger ML, Hovda DA, Giza CC. Ontogeny of rat recognition memory measured by the novel object recognition task. Dev Psychobiol 2009; 51:672-678.
31. Martins de Lima MN, Presti-Torres J, Dornelles A, Bromberg E, Schroder N. Differential effects of low and high doses of topiramate on consolidation and retrieval of novel object recognition memory in rats. Epilepsy Behav 2007; 10:32-37.
32. de Lima MN, Laranja DC, Caldana F, Bromberg E, Roesler R, Schroder N. Reversal of age-related deficits in object recognition memory in rats with l-deprenyl. Exp Gerontol 2005; 40:506-511.
33. Schroder N, O’Dell SJ, Marshall JF. Neurotoxic methamphetamine regimen severely impairs recognition memory in rats. Synapse 2003; 49:89-96.
34. Kolosowska K, Maciejak P, Szyndler J, Turzynska D, Sobolewska A, Plaznik A. The role of interleukin-1beta in the pentylenetetrazole-induced kindling of seizures, in the rat hippocampus. Eur J Pharmacol 2014; 731:31-37.
35. Hulse RE, Kunkler PE, Fedynyshyn JP, Kraig RP. Optimization of multiplexed bead-based cytokine immunoassays for rat serum and brain tissue. J Neurosci Methods 2004; 136:87-98.
36. Zhang X, Yan F, Feng J, Qian H, Cheng Z, Yang Q, et al. Dexmedetomidine inhibits inflammatory reaction in the hippocampus of septic rats by suppressing NF-kappaB pathway. PLoS One 2018; 13:e0196897.
37. Rossi O, Maggiore L, Necchi F, Koeberling O, MacLennan CA, Saul A, et al. Comparison of colorimetric assays with quantitative amino acid analysis for protein quantification of Generalized Modules for Membrane Antigens (GMMA). Mol Biotechnol 2015; 57:84-93.
38. Bagheri-Abassi F, Alavi H, Mohammadipour A, Motejaded F, Ebrahimzadeh-Bideskan A. The effect of silver nanoparticles on apoptosis and dark neuron production in rat hippocampus. Iran J Basic Med Sci 2015; 18:644-648.
39. Khazipov R, Zaynutdinova D, Ogievetsky E, Valeeva G, Mitrukhina O, Manent JB, et al. Atlas of the postnatal rat brain in stereotaxic coordinates. Front Neuroanat 2015; 9:161.
40. Pourzaki M, Homayoun M, Sadeghi S, Seghatoleslam M, Hosseini M, Ebrahimzadeh Bideskan A. Preventive effect of Coriandrum sativum on neuronal damages in pentylentetrazole-induced seizure in rats. Avicenna J Phytomed 2017; 7:116-128.
41. Zhao Y, Han Y, Bu DF, Zhang J, Li QR, Jin HF, et al. Reduced AKT phosphorylation contributes to endoplasmic reticulum stress-mediated hippocampal neuronal apoptosis in rat recurrent febrile seizure. Life Sci 2016; 153:153-162.
42. Haghir H, Hami J, Lotfi N, Peyvandi M, Ghasemi S, Hosseini M. Expression of apoptosis-regulatory genes in the hippocampus of rat neonates born to mothers with diabetes. Metab Brain Dis 2017; 32:617-628.
43. Homayoun M, Seghatoleslam M, Pourzaki M, Shafieian R, Hosseini M, Ebrahimzadeh Bideskan A. Anticonvulsant and neuroprotective effects of Rosa damascena hydro-alcoholic extract on rat hippocampus. Avicenna J Phytomed 2015; 5:260-270.
44. Wang CP, Shi YW, Tang M, Zhang XC, Gu Y, Liang XM, et al. Isoquercetin Ameliorates Cerebral Impairment in Focal Ischemia Through Anti-Oxidative, Anti-Inflammatory, and Anti-Apoptotic Effects in Primary Culture of Rat Hippocampal Neurons and Hippocampal CA1 Region of Rats. Mol Neurobiol 2017; 54:2126-2142.
45. Rajabzadeh AA, Bideskan AR, Haghir H, Fazel AR. Morphometrical study of polysialylated neural cell adhesion molecule positive cells in rat pups hippocampus following induction of seizure during pregnancy. Iran Biomed J 2011; 15:157-163.
46. Mohammadipour A, Fazel A, Haghir H, Motejaded F, Rafatpanah H, Zabihi H, et al. Maternal exposure to titanium dioxide nanoparticles during pregnancy; impaired memory and decreased hippocampal cell proliferation in rat offspring. Environ Toxicol Pharmacol 2014; 37:617-625.
47. Yagoubi N, Jomni Y, Sakly M. Hyperthermia-induced febrile seizures have moderate and transient effects on spatial learning in immature rats. Behav Neurol 2015; 2015:924303.
48. Weiss EF, Masur D, Shinnar S, Hesdorffer DC, Hinton VJ, Bonner M, et al. Cognitive functioning one month and one year following febrile status epilepticus. Epilepsy Behav 2016; 64:283-288.
49. Huang CC, Chang YC. The long-term effects of febrile seizures on the hippocampal neuronal plasticity - clinical and experimental evidence. Brain Dev 2009; 31:383-387.
50. Saeedi Borujeni MJ, Hami J, Haghir H, Rastin M, Sazegar G. Evaluation of Bax and Bcl-2 proteins expression in the rat hippocampus due to childhood febrile seizure. Iran J Child Neurol 2016; 10:53-60.
51. Reid AY, Riazi K, Campbell Teskey G, Pittman QJ. Increased excitability and molecular changes in adult rats after a febrile seizure. Epilepsia 2013; 54:e45-48.
52. Auvin S, Shin D, Mazarati A, Sankar R. Inflammation induced by LPS enhances epileptogenesis in immature rat and may be partially reversed by IL1RA. Epilepsia 2010; 51 Suppl 3:34-38.
53. Vezzani A, Aronica E, Mazarati A, Pittman QJ. Epilepsy and brain inflammation. Exp Neurol 2013; 244:11-21.
54. Kenney-Jung DL, Vezzani A, Kahoud RJ, LaFrance-Corey RG, Ho ML, Muskardin TW, et al. Febrile infection-related epilepsy syndrome treated with anakinra. Ann Neurol 2016; 80:939-945.
55. Adabi Mohazab R, Javadi-Paydar M, Delfan B, Dehpour AR. Possible involvement of PPAR-gamma receptor and nitric oxide pathway in the anticonvulsant effect of acute pioglitazone on pentylenetetrazole-induced seizures in mice. Epilepsy Res 2012; 101:28-35.
56. Okada K, Yamashita U, Tsuji S. Ameliorative effect of pioglitazone on seizure responses in genetically epilepsy-susceptible EL mice. Brain Res 2006; 1102:175-178.
57. Simeone TA, Matthews SA, Simeone KA. Synergistic protection against acute flurothyl-induced seizures by adjuvant treatment of the ketogenic diet with the type 2 diabetes drug pioglitazone. Epilepsia 2017; 58:1440-1450.
58. Martinos MM, Yoong M, Patil S, Chin RF, Neville BG, Scott RC, et al. Recognition memory is impaired in children after prolonged febrile seizures. Brain 2012; 135:3153-3164.
59. Jiang LY, Tang SS, Wang XY, Liu LP, Long Y, Hu M, et al. PPARγ agonist pioglitazone reverses memory impairment and biochemical changes in a mouse model of type 2 diabetes mellitus. CNS Neurosci Ther 2012; 18:659-666.
60. Yin QQ, Pei JJ, Xu S, Luo DZ, Dong SQ, Sun MH, et al. Pioglitazone improves cognitive function via increasing insulin sensitivity and strengthening antioxidant defense system in fructose-drinking insulin resistance rats. PLoS One 2013; 8:e59313.
61. Dube CM, Brewster AL, Richichi C, Zha Q, Baram TZ. Fever, febrile seizures and epilepsy. Trends Neurosci 2007; 30:490-496.
62. Saghazadeh A, Gharedaghi M, Meysamie A, Bauer S, Rezaei N. Proinflammatory and anti-inflammatory cytokines in febrile seizures and epilepsy: systematic review and meta-analysis. Rev Neurosci 2014; 25:281-305.
63. Dinarello CA. Infection, fever, and exogenous and endogenous pyrogens: some concepts have changed. J Endotoxin Res 2004; 10:201-222.
64. Zou JN, Xiao J, Hu SS, Fu CS, Zhang XL, Zhang ZX, et al. Toll-like receptor 4 signaling pathway in the protective effect of pioglitazone on experimental immunoglobulin A nephropathy. Chin Med J (Engl) 2017; 130:906-913.
65. Xu J, Nie M, Li J, Xu Z, Zhang M, Yan Y, et al. Effect of pioglitazone on inflammation and calcification in atherosclerotic rabbits : An (18)F-FDG-PET/CT in vivo imaging study. Herz 2017.
66. Chedrawe MAJ, Holman SP, Lamport AC, Akay T, Robertson GS. Pioglitazone is superior to quetiapine, clozapine and tamoxifen at alleviating experimental autoimmune encephalomyelitis in mice. J Neuroimmunol 2018; 321:72-82.
67. Kim S, Dede AJ, Hopkins RO, Squire LR. Memory, scene construction, and the human hippocampus. Proc Natl Acad Sci U S A 2015; 112:4767-4772.
68. Lee CH, Yi MH, Chae DJ, Zhang E, Oh SH, Kim DW. Effect of pioglitazone on excitotoxic neuronal damage in the mouse hippocampus. Biomol Ther (Seoul) 2015; 23:261-267.
69. Sauerbeck A, Gao J, Readnower R, Liu M, Pauly JR, Bing G, et al. Pioglitazone attenuates mitochondrial dysfunction, cognitive impairment, cortical tissue loss, and inflammation following traumatic brain injury. Exp Neurol 2011; 227:128-135.
70. El-Sahar AE, Safar MM, Zaki HF, Attia AS, Ain-Shoka AA. Neuroprotective effects of pioglitazone against transient cerebral ischemic reperfusion injury in diabetic rats: Modulation of antioxidant, anti-inflammatory, and anti-apoptotic biomarkers. Pharmacol Rep 2015; 67:901-906.
71. Chuang YC, Lin TK, Huang HY, Chang WN, Liou CW, Chen SD, et al. Peroxisome proliferator-activated receptors gamma/mitochondrial uncoupling protein 2 signaling protects against seizure-induced neuronal cell death in the hippocampus following experimental status epilepticus. J Neuroinflammation 2012; 9:184.
72. Offringa M, Newton R, Cozijnsen MA, Nevitt SJ. Prophylactic drug management for febrile seizures in children. Cochrane Database Syst Rev 2017; 2:CD003031.
73. Leonard CE, Han X, Brensinger CM, Bilker WB, Cardillo S, Flory JH, et al. Comparative risk of serious hypoglycemia with oral antidiabetic monotherapy: A retrospective cohort study. Pharmacoepidemiol Drug Saf 2018; 27:9-18.