Metabolic Brain Disease

, Volume 33, Issue 5, pp 1743–1753 | Cite as

The effects of repetitive stress on tat protein-induced pro-inflammatory cytokine release and steroid receptor expression in the hippocampus of rats

  • Khayelihle B. MakhathiniEmail author
  • Oualid Abboussi
  • Musa V. Mabandla
  • William M. U. Daniels
Original Article


Human immunodeficiency virus type 1 (HIV-1) affects the central nervous system (CNS) that may lead to the development of HIV-associated neuropathologies. Tat protein is one of the viral proteins that have been linked to the neurotoxic effects of HIV. Since many individuals living with HIV often experience significant adverse circumstances, the present study investigated whether exposure to stressful conditions would exacerbate harmful effects of tat protein on brain function. Tat protein (10 μg/10 μl) was injected bilaterally into the dorsal hippocampus of the animal using stereotaxic techniques. The control group received an injection of saline (10 μl). Some control and tat protein-treated animals were subjected to restrain stress for 6 h per day for 28 days and compared to a non-stress group. All animals underwent two behavioural tests, the open field test (OFT) and the novel object recognition test (NORT) to assess their mood state and cognitive function respectively. The release of pro-inflammatory cytokines (TNF-α and IL-1β) and the expression of mineralocorticoid (MR) and glucocorticoid (GR) receptors were also measured to see whether the impact of the repetitive stress on Tat protein-induced behavioural effects was mediated by elements of the immune system and the HPA axis. Rats treated with tat protein showed the following behavioural changes when compared to control animals: there was a significant decrease in time spent in the center of the open field during the OFT, a significant reduction in time spent with the novel object during the NORT, but no change in locomotor activity. Real-time PCR data showed that the expression levels of GR and MR mRNA were significantly reduced, while Western blot analysis showed that the protein expression levels of TNF-α and IL-1β were significantly increased. The present findings indicated that injection of tat protein into the hippocampus of rats not subjected to stress may lead to anxiety-like behaviour and deficits in learning and memory. Tat-treated animals subjected to stress evoked only a modest effect on their behaviour and neurochemistry, while stress alone led to behavioural and neurochemical changes similar to tat protein.


HIV-1 HIV-associated neurocognitive disorder Novel object recognition test Glucocorticoid receptors Mineralocorticoid receptors TNF-α and IL-1β 



The authors wish to thank the National Research Foundation of South Africa (NRF grant number 83792) and the University of KwaZulu-Natal for financial support, as well as the staff of the Biomedical Resource Centre of the University of KwaZulu-Natal for technical assistance.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest.


  1. Agrawal L, Louboutin JP, Reyes BA, Van Bockstaele EJ, Strayer DS (2012) HIV-1 Tat neurotoxicity: a model of acute and chronic exposure, and neuroprotection by gene delivery of antioxidant enzymes. Neurobiol Dis 45:657–670CrossRefPubMedGoogle Scholar
  2. Aleisa AM, Alzoubi KH, Gerges NZ, Alkadhi KA (2006) Chronic psychosocial stress-induced impairment of hippocampal LTP: Possible role of BDNF. Neurobiol Dis 22:453–462CrossRefPubMedGoogle Scholar
  3. Ali I, Hogberg J, Hsieh JH, Auerbach S, Korhonen A et al (2016) Gender differences in cancer susceptibility: role of oxidative stress. Carcinogenesis 37:985–992CrossRefPubMedGoogle Scholar
  4. Amin SN, El-Aidi AA, Ali MM, Attia YM, Rashed LA (2015) Modification of hippocampal markers of synaptic plasticity by memantine in animal models of acute and repeated restraint stress: implications for memory and behavior. Neuromol Med 17:121–136CrossRefGoogle Scholar
  5. Ashraf T, Jiang W, Hoque MT, Henderson J, Wu C, Bendayan R (2014) Role of anti-inflammatory compounds in human immunodeficiency virus-1 glycoprotein120-mediated brain inflammation. J Neuroinflammation 11:91CrossRefPubMedPubMedCentralGoogle Scholar
  6. Banerjee A, Zhang X, Manda KR, Banks WA, Ercal N (2010) HIV proteins (gp120 and Tat) and methamphetamine in oxidative stress-induced damage in the brain: potential role of the thiol antioxidant N-acetylcysteine amide. Free Radic Biol Med 48:1388–1398CrossRefPubMedPubMedCentralGoogle Scholar
  7. Bansal AK, Mactutus CF, Nath A, Maragos W, Hauser KF, Booze RM (2000) Neurotoxicity of HIV-1 proteins gp120 and Tat in the rat striatum. Brain Res 879:42–49CrossRefPubMedGoogle Scholar
  8. Brabers NACH, Nottet HSLM (2006) Role of the pro-inflammatory cytokines TNF-α and IL-1β in HIV-associated dementia. Eur J Clin Investig 36:447–458CrossRefGoogle Scholar
  9. Caudal D, Jay TM, Godsil BP (2014) Behavioral stress induces regionally-distinct shifts of brain mineralocorticoid and glucocorticoid receptor levels. Front Behav Neurosci 8:19CrossRefPubMedPubMedCentralGoogle Scholar
  10. Chao HM, McEwen BS (1994) Glucocorticoids and the expression of mRNAs for neurotrophins, their receptors and GAP-43 in the rat hippocampus. Brain Res Mol Brain Res 26:271–276CrossRefPubMedGoogle Scholar
  11. Cheng B, Mattson MP (1991) NGF and bFGF protect rat hippocampal and human cortical neurons against hypoglycemic damage by stabilizing calcium homeostasis. Neuron 7:1031–1041CrossRefPubMedGoogle Scholar
  12. Cheng B, Christakos S, Mattson MP (1994) Tumor necrosis factors protect neurons against metabolic-excitotoxic insults and promote maintenance of calcium homeostasis. Neuron 12:139–153CrossRefPubMedGoogle Scholar
  13. Chittiprol S, Shetty KT, Kumar AM, Bhimasenarao RS, Satishchandra P et al (2007) HPA axis activity and neuropathogenesis in HIV-1 clade C infection. Front Biosci 12:1271–1277CrossRefPubMedGoogle Scholar
  14. Cole SW (2008) Psychosocial influences on HIV-1 disease progression: neural, endocrine, and virologic mechanisms. Psychosom Med 70:562–568CrossRefPubMedGoogle Scholar
  15. Daniels WM, Jaffer A, Engelbrecht AH, Russell VA, Taljaard JJ (1990) The effect of intrahippocampal injection of kainic acid on corticosterone release in rats. Neurochem Res 15:495–499CrossRefPubMedGoogle Scholar
  16. Daniels WM, Pietersen CY, Carstens ME, Stein DJ (2004a) Maternal separation in rats leads to anxiety-like behavior and a blunted ACTH response and altered neurotransmitter levels in response to a subsequent stressor. Metab Brain Dis 19:3–14CrossRefPubMedGoogle Scholar
  17. Daniels WM, Richter L, Stein DJ (2004b) The effects of repeated intra-amygdala CRF injections on rat behavior and HPA axis function after stress. Metab Brain Dis 19:15–23CrossRefPubMedGoogle Scholar
  18. de Rezende MG, Garcia-Leal C, de Figueiredo FP, Cavalli RC, Spanghero MS et al (2016) Altered functioning of the HPA axis in depressed postpartum women. J Affect Disord 193:249–256CrossRefPubMedGoogle Scholar
  19. Dean RL 3rd, Scozzafava J, Goas JA, Regan B, Beer B, Bartus RT (1981) Age-related differences in behavior across the life span of the C57BL/6J mouse. Exp Aging Res 7:427–451CrossRefPubMedGoogle Scholar
  20. Ennaceur A (2010) One-trial object recognition in rats and mice: methodological and theoretical issues. Behav Brain Res 215:244–254CrossRefPubMedGoogle Scholar
  21. Faure A, Naldi A, Chaouiya C, Thieffry D (2006) Dynamical analysis of a generic Boolean model for the control of the mammalian cell cycle. BMC (Oxford, England) 22:e124–e131CrossRefGoogle Scholar
  22. Fumaz CR, Gonzalez-Garcia M, Borras X, Muñoz-Moreno JA, Perez-Alvarez N et al (2012) Psychological stress is associated with high levels of IL-6 in HIV-1 infected individuals on effective combined antiretroviral treatment. Brain Behav Immun 26:568–572CrossRefPubMedGoogle Scholar
  23. Gądek-Michalska A, Bugajski J (2010) Interleukin-1 (IL-1) in stress-induced activation of limbic-hypothalamic-pituitary adrenal axis. Pharmacol Rep 62(6):969–982CrossRefPubMedGoogle Scholar
  24. Grant I (2008) Neurocognitive disturbances in HIV. Int Rev Psychiatry (Abingdon, England) 20:33–47CrossRefGoogle Scholar
  25. Guedia J, Brun P, Bhave S, Fitting S, Kang M et al (2016) HIV-1 Tat exacerbates lipopolysaccharide-induced cytokine release via TLR4 signaling in the enteric nervous system. Sci Rep 6:31203CrossRefPubMedPubMedCentralGoogle Scholar
  26. Harricharan R, Thaver V, Russell VA, Daniels WM (2015) Tat-induced histopathological alterations mediate hippocampus-associated behavioural impairments in rats. Behav Brain Funct: BBF 11:3CrossRefPubMedGoogle Scholar
  27. Heaton RK, Clifford DB, Franklin DR, Woods SP, Ake C et al (2010) HIV-associated neurocognitive disorders persist in the era of potent antiretroviral therapy: CHARTER Study. Neurology 75:2087–2096CrossRefPubMedPubMedCentralGoogle Scholar
  28. Howland LC, Gortmaker SL, Mofenson LM, Spino C, Gardner JD et al (2000) Effects of negative life events on immune suppression in children and youth infected with human immunodeficiency virus type 1. Pediatrics 106:540–546CrossRefPubMedGoogle Scholar
  29. Jurgens HA, Johnson RW (2012) Environmental enrichment attenuates hippocampal neuroinflammation and improves cognitive function during influenza infection. Brain Behav Immun 26:1006–1016CrossRefPubMedPubMedCentralGoogle Scholar
  30. Kasahara E, Inoue M (2015) Cross-talk between HPA-axis-increased glucocorticoids and mitochondrial stress determines immune responses and clinical manifestations of patients with sepsis. Redox Report: Free Radic Res 20:1–10CrossRefGoogle Scholar
  31. Kilkenny C, Browne W, Cuthill IC, Emerson M, Altman DG (2010) Animal research: Reporting in vivo experiments: The ARRIVE guidelines. Br J Pharmacol 160:1577–1579CrossRefPubMedPubMedCentralGoogle Scholar
  32. Kirby ED, Muroy SE, Sun WG, Covarrubias D, Leong MJ et al (2013) Acute stress enhances adult rat hippocampal neurogenesis and activation of newborn neurons via secreted astrocytic FGF2. eLife 2:e00362CrossRefPubMedPubMedCentralGoogle Scholar
  33. Kruman II, Nath A, Mattson MP (1998) HIV-1 protein Tat induces apoptosis of hippocampal neurons by a mechanism involving caspase activation, calcium overload, and oxidative stress. Exp Neurol 154:276–288CrossRefPubMedGoogle Scholar
  34. Kumar M, Kumar AM, Waldrop D, Antoni MH, Eisdorfer C (2003) HIV-1 infection and its impact on the HPA axis, cytokines, and cognition. Stress (Amsterdam, Netherlands) 6:167–172CrossRefGoogle Scholar
  35. Lawson MA, Kelley KW, Dantzer R (2011) Intracerebroventricular administration of HIV-1 Tat induces brain cytokine and indoleamine 2,3-dioxygenase expression: a possible mechanism for AIDS comorbid depression. Brain Behav Immun 25:1569–1575CrossRefPubMedPubMedCentralGoogle Scholar
  36. Leserman J (2003) HIV disease progression: depression, stress, and possible mechanisms. Biol Psychiatry 54:295–306CrossRefPubMedGoogle Scholar
  37. Leserman J (2008) Role of depression, stress, and trauma in HIV disease progression. Psychosom Med 70:539–545CrossRefPubMedGoogle Scholar
  38. Lewis CF (2005) Post-traumatic stress disorder in HIV-positive incarcerated women. J Am Acad Psychiatry Law 33:455–464PubMedGoogle Scholar
  39. Lindberg C, Selenica ML, Westlind-Danielsson A, Schultzberg M (2005) Beta-amyloid protein structure determines the nature of cytokine release from rat microglia. J Mol Neurosci: MN 27:1–12CrossRefPubMedGoogle Scholar
  40. Lupien SJ, McEwen BS, Gunnar MR, Heim C (2009) Effects of stress throughout the lifespan on the brain, behaviour and cognition. Nat Rev Neurosci 10:434CrossRefPubMedGoogle Scholar
  41. Makhathini KB, Abboussi O, Stein DJ, Mabandla MV, Daniels WMU (2017) Repetitive stress leads to impaired cognitive function that is associated with DNA hypomethylation, reduced BDNF and a dysregulated HPA axis. Int J Dev Neurosci 60:63–69CrossRefPubMedGoogle Scholar
  42. McEwen BS (2007) Physiology and neurobiology of stress and adaptation: central role of the brain. Physiol Rev 87:873–904CrossRefPubMedGoogle Scholar
  43. Membreno L, Irony I, Dere W, Klein R, Biglieri EG, Cobb E (1987) Adrenocortical function in acquired immunodeficiency syndrome. J Clin Endocrinol Metab 65:482–487CrossRefPubMedGoogle Scholar
  44. Nicoli F, Finessi V, Sicurella M, Rizzotto L, Gallerani E et al (2013) The HIV-1 Tat protein induces the activation of CD8+ T cells and affects in vivo the magnitude and kinetics of antiviral responses. PLoS One 8:e77746CrossRefPubMedPubMedCentralGoogle Scholar
  45. Ogłodek E, Szota A, Just M, Moś D, Araszkiewicz A (2014) The role of the neuroendocrine and immune systems in the pathogenesis of depression. Pharmacol Rep 66(5):776–781CrossRefPubMedGoogle Scholar
  46. Paul D, Madan V, Bartenschlager R (2014) Hepatitis C virus RNA replication and assembly: living on the fat of the land. Cell Host Microbe 16:569–579CrossRefPubMedGoogle Scholar
  47. Paxinos G, Watson C, Pennisi M, Topple A (1985) Bregma, lambda and the interaural midpoint in stereotaxic surgery with rats of different sex, strain and weight. J Neurosci Methods 13(2):139–143CrossRefPubMedGoogle Scholar
  48. Planes R, Ben Haij N, Leghmari K, Serrero M, BenMohamed L, Bahraoui E (2016) HIV-1 Tat Protein Activates both the MyD88 and TRIF Pathways To Induce Tumor Necrosis Factor Alpha and Interleukin-10 in Human Monocytes. J Virol 90:5886–5898CrossRefPubMedPubMedCentralGoogle Scholar
  49. Puccini JM, Marker DF, Fitzgerald T, Barbieri J, Kim CS et al (2015) Leucine-rich repeat kinase 2 modulates neuroinflammation and neurotoxicity in models of human immunodeficiency virus 1-associated neurocognitive disorders. J Neurosci 35:5271–5283CrossRefPubMedPubMedCentralGoogle Scholar
  50. Ramautar A, Mabandla M, Blackburn J, Daniels WMU (2012) Inhibition of HIV-1 tat-induced transactivation and apoptosis by the divalent metal chelators, fusaric acid and picolinic acid—Implications for HIV-1 dementia. Neurosci Res 74:59–63CrossRefPubMedGoogle Scholar
  51. Roth KA, Katz RJ (1979) Stress, behavioral arousal, and open field activity--a reexamination of emotionality in the rat. Neurosci Biobehav Rev 3:247–263CrossRefPubMedGoogle Scholar
  52. Royal W, Cherner M, Carr J, Habib AG, Akomolafe A et al (2012) Clinical Features and Virological Correlates of Neurocognitive Impairment among HIV-Infected Individuals in Nigeria. J Neurovirol 18:191–199CrossRefPubMedPubMedCentralGoogle Scholar
  53. Rubinstein PG, Aboulafia DM, Zloza A (2014) Malignancies in HIV/AIDS: From Epidemiology to Therapeutic Challenges. AIDS (London, England) 28:453–465CrossRefGoogle Scholar
  54. Saadat M, Behboodi ZM, Saadat E (2015) Comparison of depression, anxiety, stress, and related factors among women and men with human immunodeficiency virus infection. J Hum Reprod Sci 8:48–51CrossRefPubMedPubMedCentralGoogle Scholar
  55. Shih R-H, Wang C-Y, Yang C-M (2015) NF-kappaB signaling pathways in neurological inflammation: a mini review. Front Mol Neurosci 8:77CrossRefPubMedPubMedCentralGoogle Scholar
  56. Singh D, Joska JA, Goodkin K, Lopez E, Myer L et al (2008) Normative scores for a brief neuropsychological battery for the detection of HIV-associated neurocognitive disorder (HAND) among South Africans. BMC Res Notes 3:28CrossRefGoogle Scholar
  57. Sterley TL, Howells FM, Russell VA (2016) Genetically determined differences in noradrenergic function: The spontaneously hypertensive rat model. Brain Res 1641:291–305CrossRefPubMedGoogle Scholar
  58. Uys JD, Marais L, Faure J, Prevoo D, Swart P et al (2006) Developmental trauma is associated with behavioral hyperarousal, altered HPA axis activity, and decreased hippocampal neurotrophin expression in the adult rat. Ann N Y Acad Sci 1071:542–546CrossRefPubMedGoogle Scholar
  59. Valdez AN, Rubin LH, Neigh GN (2016) Untangling the Gordian knot of HIV, stress, and cognitive impairment. Neurobiology of Stress 10:1–11Google Scholar
  60. Viel J, McManus D, Smith SS, Brewer G (2001) Age-and concentration-dependent neuroprotection and toxicity by TNF in cortical neurons from β-amyloid. J Neurosci Res 64:454–465CrossRefPubMedGoogle Scholar
  61. Wang P, Barks JD, Silverstein FS (1999) Tat, a human immunodeficiency virus-1-derived protein, augments excitotoxic hippocampal injury in neonatal rats. Neuroscience 88:585–597CrossRefPubMedGoogle Scholar
  62. Wrona D, Listowska M, Kubera M, Glac W, Grembecka B et al (2014) Effects of chronic desipramine pretreatment on open field-induced suppression of blood natural killer cell activity and cytokine response depend on the rat's behavioral characteristics. J Neuroimmunol 268:13–24CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Human Physiology, College of Health SciencesUniversity of KwaZulu-NatalDurbanSouth Africa
  2. 2.School of PhyisiologyUniversity of the WitwatersrandJohannesburgSouth Africa

Personalised recommendations