Improved spatial memory, neurobehavioral outcomes, and neuroprotective effect after progesterone administration in ovariectomized rats with traumatic brain injury: Role of RU486 progesterone receptor antagonist

Document Type : Original Article


1 Neuroscience Research and Physiology Research Centers, Kerman University of Medical Sciences, Kerman, Iran

2 Endocrinology and Metabolism Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran

3 Neuroscience Research Center, Kerman University of Medical Sciences, Kerman, Iran

4 Physiology Research Centers, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran

5 Department of Physiology, Bam University of Medical Sciences, Bam, Iran

6 Department of Psychology, Genetic Institute, Islamic Azad University- Zarand Branch, Kerman, Iran


Objective(s): The contribution of classic progesterone receptors (PR) in interceding the neuroprotective efficacy of progesterone (P4) on the prevention of brain edema and long-time behavioral disturbances was assessed in traumatic brain injury (TBI).
Materials and Methods: Female Wistar rats were ovariectomized and apportioned into 6 groups: sham, TBI, oil, P4, vehicle, and RU486. P4 or oil was injected following TBI. The antagonist of PR (RU486) or DMSO was administered before TBI. The brain edema and destruction of the blood-brain barrier (BBB) were determined. Intracranial pressure (ICP), cerebral perfusion pressure (CPP), and beam walk (BW) task were evaluated previously and at various times post-trauma. Long-time locomotor and cognitive consequences were measured one day before and on days 3, 7, 14, and 21 after the trauma.
Results: RU486 eliminated the inhibitory effects of P4 on brain edema and BBB leakage (p <0.05, p <0.001, respectively). RU486 inhibited the decremental effect of P4 on ICP as well as the increasing effect of P4 on CPP (p <0.001) after TBI. Also, RU486 inhibited the effect of P4 on the increase in traversal time and reduction in vestibulomotor score in the BW task (p <0.001). TBI induced motor, cognitive, and anxiety-like disorders, which lasted for 3 weeks after TBI; but, P4 prevented these cognitive and behavioral abnormalities (p <0.05), and RU486 opposed this P4 effect (p <0.001).
Conclusion: The classic progesterone receptors have neuroprotective effects and prevent long-time behavioral and memory deficiency after brain trauma.


1. O’Connor WT, Smyth A, Gilchrist MD. Animal models of traumatic brain injury: A critical evaluation. Pharmacol Ther 2011;130:106–113.
2. Djebaili M, Hoffman SW, Stein DG. Allopregnanolone and progesterone decrease cell death and cognitive deficits after a contusion of the rat pre-frontal cortex. Neuroscience 2004;123:349–359.
3. Vakil E. The effect of moderate to severe traumatic brain injury (tbi) on different aspects of memory: A selective review. J Clin Exp Neuropsychol  2005;27:977–1021.
4. Albensi BC, Janigro D. Traumatic brain injury and its effects on synaptic plasticity. Brain Inj 2003;17:653–663.
5. Ariza M, Serra-Grabulosa JM, Junqué C, Ramírez B, Mataró M, Poca A, et al. Hippocampal head atrophy after traumatic brain injury. Neuropsychologia 2006;44:1956–1961.
6. Hicks RR, Smith DH, Lowenstein DH, Marier Saint Mcintosh TK. Mild experimental brain injury in the rat induces cognitive deficits associated with regional neuronal loss in the hippocampus. J Neurotrauma 1993;10:405–414.
7. Baykara B, Aksu I, Buyuk E, Kiray M, Sisman AR, Baykara B, et al. Progesterone treatment decreases traumatic brain injury induced anxiety and is correlated with increased serum IGF-1 levels; Prefrontal cortex, amygdala, hippocampus neuron density; And reduced serum corticosterone levels in immature rats. Biotech Histochem 2013;88:250–257.
8. Webster KM, Wright DK, Sun M, Semple BD, Ozturk E, Stein DG, et al. Progesterone treatment reduces neuroinflammation, oxidative stress and brain damage and improves long-term outcomes in a rat model of repeated mild traumatic brain injury. J Neuroinflammation 2015;12:238-250.
9. Grossman KJ, Goss CW, Stein DG. Effects of progesterone on the inflammatory response to brain injury in the rat. Brain Res 2004;1008:29–39.
10. Khaksari M, Soltani Z, Shahrokhi N, Moshtaghi G, Asadikaram G. The role of estrogen and progesterone, administered alone and in combination, in modulating cytokine concentration following traumatic brain injury. Can J Physiol Pharmacol 2011;89:31–40.
11. Sarkaki AR, Khaksari Haddad M, Soltani Z, Shahrokhi N, Mahmoodi M. Time- and dose-dependent neuroprotective effects of sex steroid hormones on inflammatory cytokines after a traumatic brain injury. J Neurotrauma 2011;30:47–54.
12. Liu R, Wen Y, Perez E, Wang X, Day AL, Simpkins JW, et al. 17 beta-estradiol attenuates blood-brain barrier disruption induced by cerebral ischemia-reperfusion injury in female rats. Brain Res 2005;1060:55–61.
13. Soltani Z, Khaksari M, Shahrokhi N, Mohammadi G, Mofid B, Vaziri A, et al. Effect of estrogen and/or progesterone administration on traumatic brain injury-caused brain edema: the changes of aquaporin-4 and interleukin-6. J Physiol Biochem 2016;72:33–44.
14. Chang C-M, Su Y-F, Chang C-Z, Chung C-L, Tsai Y-J, Loh J-K, et al. Progesterone attenuates experimental subarachnoid hemorrhage-induced vasospasm by upregulation of endothelial nitric oxide synthase via akt signaling pathway. Biomed Res 2014;2014:1–6.
15. Roof RL, Hoffman SW, Stein DG. Progesterone protects against lipid peroxidation following traumatic brain injury in rats. Mol Chem Neuropathol 1997;31:1–11.
16. Si D, Li J, Liu J, Wang X, Wei Z, Tian Q, et al. Progesterone protects blood ‑ brain barrier function and improves neurological outcome following traumatic brain injury in rats. Exp Ther 2014;8:1010–1014.
17. Roof RL, Duvdevani R, Braswell L, Stein DG. Progesterone facilitates cognitive recovery and reduces secondary neuronal loss caused by cortical contusion injury in male rats. Exp Neurol 1994;129:64–69.
18. Petersen SL, Intlekofer KA, Moura-Conlon PJ, Brewer DN, del Pino Sans J, Lopez JA. Nonclassical progesterone signalling molecules in the nervous system. J Neuroendocrinol 2013;25:991–1001.
19. Bali N, Arimoto JM, Iwata N, Lin SW, Zhao L, Brinton RD, et al. Differential responses of progesterone receptor membrane component-1 (pgrmc1) and the classical progesterone receptor (pgr) to 17β-estradiol and progesterone in hippocampal subregions that support synaptic remodeling and neurogenesis. Endocrinology  2012;153:759–769.
20. Zhu X, Fréchou M, Schumacher M, Guennoun R. Cerebroprotection by progesterone following ischemic stroke: Multiple effects and role of the neural progesterone receptors. J Steroid Biochem Mol Biol  2019;185:90–102.
21. Ishihara Y, Fujitani N, Sakurai H, Takemoto T, Ikeda-Ishihara N, Mori-Yasumoto K, et al. Effects of sex steroid hormones and their metabolites on neuronal injury caused by oxygen-glucose deprivation/reoxygenation in organotypic hippocampal slice cultures. Steroids  2016;113:71–77.
22. Maghool F, siahposht khachki , Mohammad Khaksari A. Differences in brain edema and intracranial pressure following traumatic brain injury across the estrous cycle: Involvement of female sex steroid hormones. Brain Res 2013;1497:61–72.
23. Crandall C, Palla S, Reboussin B, Hu P, Barrett-Connor E, Reuben D, et al. Cross-sectional association between markers of inflammation and serum sex steroid levels in the postmenopausal estrogen/progestin interventions trial. J Women’s Heal [Internet]. 2006;15:14–23.
24. Sehajpal J, Kaur T, Bhatti R, Singh AP. Role of progesterone in melatonin-mediated protection against acute kidney injury. J Surg Res 2014;191:441–447.
25. Marmarou A, Abd-Elfattah Foda MA, Van den Brink W, Campbell J, Kita H, Demetriadou K. A new model of diffuse brain injury in rats. Part I: Pathophysiology and biomechanics. J Neurosurg 1994;80:291–300.
26. Naderi V, Khaksari M, Abbasi R, Maghool F. Estrogen provides neuroprotection against brain edema and blood brain barrier disruption through both estrogen receptors α and β following traumatic brain injury. Iran J Basic Med Sci 2015;18:138–144.
27. Khaksari M, Abbasloo E, Dehghan F, Soltani Z, Asadikaram G. The brain cytokine levels are modulated by estrogen following traumatic brain injury: Which estrogen receptor serves as modulator? Int Immunopharmacol 2015;28:279–287.
28. Khaksari M, Mahmmodi R, Shahrokhi N, Shabani M, Joukar S, Aqapour M. The effects of shilajit on brain edema, intracranial pressure and neurologic outcomes following the traumatic brain injury in rat. Iran J Basic Med Sci 2013;16:858–864.
29. Gilkes CE, Whitfield PC. Intracranial pressure and cerebral blood flow. A pathophysiological and clinical perspective. Surg 2009;27:139–144.
30. Soltani Z, Khaksari M, Jafari E, Iranpour M, Shahrokhi N. Is genistein neuroprotective in traumatic brain injury?. Physiology & Behavior 2015;152:26–31.
31. Vink R, O’Connor CA, Nimmo AJ, Heath DL. Magnesium attenuates persistent functional deficits following diffuse traumatic brain injury in rats. Neurosci Lett. 2003;336:41–44.
32. Bruschetta G, Impellizzeri D, Campolo M, Casili G, Di Paola R, Paterniti I, et al. FeTPPS reduces secondary damage and improves neurobehavioral functions after traumatic brain injury. Front Neurosci  2017;11:1-13.
33. Bashiri H, Rezayof A, Sahebgharani M, Tavangar SM, Zarrindast MR. Modulatory effects of the basolateral amygdala α2-adrenoceptors on nicotine-induced anxiogenic-like behaviours of rats in the elevated plus maze. Neuropharmacology 2016;105:478–86.
34. Amirkhosravi L, Maryam Raoof, Ramin Raoof, MA, Mahani SE, Ramazani M, Hamed Ebrahim nejad SA, Haghani J. Is inflammatory pulpal pain a risk factor for amnesia? Iran J Vet Sci Technol 2015;6:62–76.
35. Schwarzbach E, Bonislawski DP, Xiong G, Cohen AS. Mechanisms underlying the inability to induce area CA1 LTP in the mouse after traumatic brain injury. Hippocampus 2006;16:541–550.
36. Unterberg AW, Stover J, Kress B, Kiening KL. Edema and brain trauma. Neuroscience 2004;129:1019–1027.
37. Gabrielian L, Willshire LW, Helps SC, van den Heuvel C, Mathias J, Vink R. Intracranial pressure changes following traumatic brain injury in rats: Lack of significant change in the absence of mass lesions or hypoxia. J Neurotrauma  2011 ;28:2103–2111.
38. Bramlett HM, Dietrich WD. Neuropathological protection after traumatic brain injury in intact female rats versus males or ovariectomized females. J Neurotrauma 2001;18:891–900.
39. Shahrokhi N, Khaksari M, Soltani Z, Mahmoodi M, Nakhaee N. Effect of sex steroid hormones on brain edema, intracranial pressure, and neurologic outcomes after traumatic brain injury. Can J Physiol Pharmacol 2010;88:414–421.
40. Wang X, Zhang J, Yang Y, Dong W, Wang F, Wang L, et al. Progesterone attenuates cerebral edema in neonatal rats with hypoxic-ischemic brain damage by inhibiting the expression of matrix metalloproteinase-9 and aquaporin-4. Exp Ther Med 2013;6:263–267.
41. Feng Y, Bhatt AJ. Corticosteroid responses following hypoxic preconditioning provide neuroprotection against subsequent hypoxic-ischemic brain injury in the newborn rats. 2015;44:6–13.
42. Zhang Z, Yang R, Cai W, Bai Y, Sokabe M, Chen L. Treatment with progesterone after focal cerebral ischemia suppresses proliferation of progenitor cells but enhances survival of newborn neurons in adult male mice. Neuropharmacology 2010;58:930–939.
43. Benzel EC, Gelder FB. Correlation between sex hormone binding and peritumoral edema in intracranial meningiomas. Neurosurgery 1988;23:169–174.
44. Kawoos U, McCarron RM, Auker CR, Chavko M. Advances in intracranial pressure monitoring and its significance in managing traumatic brain injury. Int J Mol Sci 2015;16;28979–28997.
45. Marmarou A. The pathophysiology of brain edema and elevated intracranial pressure. Cleve Clin J Med  2004;71:S6-S8.
46. Marmarou A, Signoretti S, Fatouros PP, Portella G, Aygok GA, Bullock MR. Predominance of cellular edema in traumatic brain swelling in patients with severe head injuries. J Neurosurg  2008;104:720–730.
47. Neal CJ, Lee EY, Gyorgy A, Ecklund JM, Agoston DV, Ling GSF. Effect of penetrating brain injury on aquaporin-4 expression using a rat model. J Neurotrauma 2007;24:1609–1617.
48. Engelborghs K, Verlooy J, Van Reempts J, Van Deuren B, Van de Ven M, Borgers M. Temporal changes in intracranial pressure in a modified experimental model of closed head injury. J Neurosurg  2009;89:796–806.
49. Wise PM, Dubal DB, Wilson ME, Rau SW, Liu Y. Estrogens: Trophic and protective factors in the adult brain. Front Neuroendocrinol 2001;22:33-66.
50. Krause DN, Geary GG, McNeill AM, Ospina J, Duckles SP. Impact of hormones on the regulation of cerebral vascular tone. Int Congr Ser 2002;1235:395–399.
51. He J, Evans CO, Hoffman SW, Oyesiku NM, Stein DG. Progesterone and allopregnanolone reduce inflammatory cytokines after traumatic brain injury. Exp Neurol 2004;189:404–412.
52. Marik P, Chen K, Varon J, Fromm R, Sternbach GL. Management of increased intracranial pressure: A review for clinicians. J Emerg Med 1999;17:711–719.
53. Kawa L, Arborelius UP, Yoshitake T, Kehr J, Hökfelt T, Risling M, et al. Neurotransmitter systems in a mild blast traumatic brain injury model: catecholamines and serotonin. J Neurotrauma  2014;32:1190–1199.
54. Washington PM, Forcelli PA, Wilkins T, Zapple DN, Parsadanian M, Burns MP. The effect of injury severity on behavior: a phenotypic study of cognitive and emotional deficits after mild, moderate, and severe controlled cortical impact injury in mice. J Neurotrauma 2012;29:2283–2296.
55. Zohar O, Rubovitch V, Milman A, Schreiber S, Pick CG. Behavioral consequences of minimal traumatic brain injury in mice. Acta Neurobiol Exp 2011;71:36–45.
56. McCullers DL, Sullivan PG, Scheff SW, Herman JP. Traumatic brain injury regulates adrenocorticosteroid receptor mRNA levels in rat hippocampus. Brain Res 2002;947:41–49.
57. Woolf PD, Cox C, Kelly M, Nichols D, McDonald J V., Hamill RW. The adrenocortical response to brain injury: Correlation with the severity of neurologic dysfunction, effects of intoxication, and patient outcome. Alcohol Clin Exp Res 1990;14:917–921.
58. Walter B, Brust P, Füchtner F, Müller M, Hinz R, Kuwabara H, et al. Age-dependent effects of severe traumatic brain injury on cerebral dopaminergic activity in newborn and juvenile pigs. J Neurotrauma  2004;21:1076–1089.
59. Sugo N, Hurn PD, Morahan MB, Hattori K, Traystman RJ, Devries AC. Social stress exacerbates focal cerebral ischemia in mice. Stroke 2002;33:1660-1664.
60. Geddes RI, Peterson BL, Stein DG, Sayeed I. Progesterone treatment shows benefit in female rats in a pediatric model of controlled cortical impact injury. McCormick CM, editor. PLoS One 2016;11:e0146419.
61. Frye CA, Walf AA, Rhodes ME, Harney JP. Progesterone enhances motor, anxiolytic, analgesic, and antidepressive behavior of wild-type mice, but not those deficient in type 1 5α-reductase. Brain Res 2004;1004:116–124.
62. Guennoun R, Zhu X, Fréchou M, Gaignard P, Slama A, Liere P, et al. Steroids in stroke with special reference to progesterone. Cell Mol Neurobiol 2018;39:551-568.
63. An C, Jiang X, Pu H, Hong D, Zhang W, Hu X, et al. Severity-dependent long-term spatial learning-memory impairment in a mouse model of traumatic brain injury. Transl Stroke Res  2016;7:512–520.
64. PanelClaudiaEspinosa-García, AlejandraAguilar-Hernández, Miguel Cervantes G. Effects of progesterone on neurite growth inhibitors in the hippocampus following global cerebral ischemia. Brain Res 2014;1545:23–34.
65. Li XJ, He RF, Li S, Li XJ, Li DL. Effects of progesterone on learning and memory and P2X7 receptor expression in the hippocampus after global cerebral ischemia/ reperfusion injury in rats. Chinese J Appl Physiol  2012;28:472–475.
66. Espinosa-García C, Aguilar-Hernández A, Cervantes M, Moralí G. Effects of progesterone on neurite growth inhibitors in the hippocampus following global cerebral ischemia. Brain Res 2014;1545:23–34.
67. Newell AJ, Lalitsasivimol D, Willing J, Gonzales K, Waters EM, Milner TA, et al. Progesterone receptor expression in cajal-retzius cells of the developing rat dentate gyrus: Potential role in hippocampus-dependent memory. J Comp Neurol 2018;526:2285–2300.
68. Velosky AG, Tucker LB, Fu AH, Liu J, McCabe JT. Cognitive performance of male and female C57BL/6J mice after repetitive concussive brain injuries. Behav Brain Res 2017;324:115–24.
69. Marschner L, Schreurs A, Lechat B, Mogensen J, Roebroek A, Ahmed T, et al. Single mild traumatic brain injury results in transiently impaired spatial long-term memory and altered search strategies. Behav Brain Res 2019;365:222–230.
70. Barha CK, Ishrat T, Epp JR, Galea LAM, Stein DG. Progesterone treatment normalizes the levels of cell proliferation and cell death in the dentate gyrus of the hippocampus after traumatic brain injury. Exp Neurol  2011;231:72–81.
71. Li X, Zhang J, Zhu X, Hou R, Li X, Dong X, et al. Effects of progesterone on hippocampal ultrastructure and expression of inflammatory mediators in neonatal rats with hypoxic-ischemic brain injury. Exp Ther Med 2014;7:1311–1316.
72. Uysal N, Baykara B, Kiray M, Cetin F, Aksu I, Dayi A, et al. Combined treatment with progesterone and magnesium sulfate positively affects traumatic brain injury in immature rats. Turk Neurosurg 2013;23:129–137.
73. Jiang C, Zuo F, Wang Y, Lu H, Yang Q, Wang J. Progesterone changes VEGF and BDNF expression and promotes neurogenesis after ischemic stroke. Mol Neurobiol 2017;54:571–581.
74. Cutler SM, Cekic M, Miller DM, Wali B, VanLandingham JW, Stein DG. Progesterone improves acute recovery after traumatic brain injury in the aged rat. J Neurotrauma  2007;24:1475–1486.
75. Cai W, Zhu Y, Furuya K, Li Z, Sokabe M, Chen L. Two different molecular mechanisms underlying progesterone neuroprotection against ischemic brain damage. Neuropharmacology 2008;55:127–138.
76. Kadhim HJ, Duchateau J, Sébire G. Cytokines and brain injury: Invited review. J Intensive Care Med 2008;23:236–249.
77. Djebaili M, Guo Q, Pettus EH, Hoffman SW, Stein DG. The neurosteroids progesterone and allopregnanolone reduce cell death, gliosis, and functional deficits after traumatic brain injury in rats. J Neurotrauma 2005;22:106–118.
78. Zhu X, Fréchou M, Liere P, Zhang S, Pianos A, Fernandez N, et al. A role of endogenous progesterone in stroke cerebroprotection revealed by the neural-specific deletion of its intracellular receptors. J Neurosci 2017;37:10998–11020.
79. Lei B, Mace B, Dawson HN, Warner DS, Laskowitz DT, James ML. Anti-inflammatory effects of progesterone in lipopolysaccharide-stimulated BV-2 microglia. PLoS One 2014;9:3–9.
80. Jodhka PK, Kaur P, Underwood W, Lydon JP, Singh M. The differences in neuroprotective efficacy of progesterone and medroxyprogesterone acetate correlate with their effects on brain-derived neurotrophic factor expression. Endocrinology 2009;150:3162–3168.
81. Singh M, Su C, Ng S. Non-genomic mechanisms of progesterone action in the brain. Front Neurosci 2013;7:159-175.
82. Luoma JI, Kelley BG, Mermelstein PG. Progesterone inhibition of voltage-gated calcium channels is a potential neuroprotective mechanism against excitotoxicity. Steroids 2011;76:845–855.