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Activation of the inflammatory transcription factor nuclear factor interleukin-6 during inflammatory and psychological stress in the brain.

Fuchs F, Damm J, Gerstberger R, Roth J, Rummel C - J Neuroinflammation (2013)

Bottom Line: In vitro NF-IL6-activation was associated with induction and secretion of TNFα in folliculostellate cells, which could be antagonized by the JAK-STAT-inhibitor AG490.Both stressors were associated with time-dependent activation of NF-IL6 in different cell types of the brain and the pituitary.Moreover, while NF-IL6-IR was partially linked to STAT3 and NFκB activation, TNFα production, and ACTH-IR after LPS stimulation; this was not the case after exposure to a novel environment, suggesting distinct underlying signaling pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Veterinary-Physiology and -Biochemistry, Justus-Liebig-University Giessen, Frankfurter Strasse 100, Giessen D-35392, Germany. Christoph.D.Rummel@vetmed.uni-giessen.de.

ABSTRACT

Background: The transcription factor nuclear factor interleukin 6 (NF-IL6) is known to be activated by various inflammatory stimuli in the brain. Interestingly, we recently detected NF-IL6-activation within the hypothalamus-pituitary-adrenal (HPA)-axis of rats after systemic lipopolysaccharide (LPS)-injection. Thus, the aim of the present study was to investigate whether NF-IL6 is activated during either, inflammatory, or psychological stress in the rat brain.

Methods: Rats were challenged with either the inflammatory stimulus LPS (100 μg/kg, i.p.) or exposed to a novel environment. Core body temperature (Tb) and motor activity were monitored using telemetry, animals were killed at different time points, brains and blood removed, and primary cell cultures of the anterior pituitary lobe (AL) were investigated. Analyses were performed using immunohistochemistry, RT-PCR, and cytokine-specific bioassays.

Results: Stress stimulation by a novel environment increased NF-IL6-immunoreactivity (IR) in the pituitary's perivascular macrophages and hypothalamic paraventricular cells and a rise in Tb lasting approximately 2 h. LPS stimulation lead to NF-IL6-IR in several additional cell types including ACTH-IR-positive corticotrope cells in vivo and in vitro. Two other proinflammatory transcription factors, namely signal transducer and activator of transcription (STAT)3 and NFκB, were significantly activated and partially colocalized with NF-IL6-IR in cells of the AL only after LPS-stimulation, but not following psychological stress. In vitro NF-IL6-activation was associated with induction and secretion of TNFα in folliculostellate cells, which could be antagonized by the JAK-STAT-inhibitor AG490.

Conclusions: We revealed, for the first time, that NF-IL6 activation occurs not only during inflammatory LPS stimulation, but also during psychological stress, that is, a novel environment. Both stressors were associated with time-dependent activation of NF-IL6 in different cell types of the brain and the pituitary. Moreover, while NF-IL6-IR was partially linked to STAT3 and NFκB activation, TNFα production, and ACTH-IR after LPS stimulation; this was not the case after exposure to a novel environment, suggesting distinct underlying signaling pathways. Overall, NF-IL6 can be used as a broad activation marker in the brain and might be of interest for therapeutic approaches not only during inflammatory but also psychological stress.

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Tb, motor activity, and plasma IL-6 levels during novel environment stress or control situation in rats. (A) Novel environment stress (stress) induced a rise in body temperature (Tb). (B) This response was preceded and accompanied by increased motor activity (averaged cumulative activity over 15 min for each) but (C) no significant rise in plasma interleukin 6 (IL-6)-levels, when compared over time to unstressed control animals. Dashed black arrows indicate changes in the number of animals; this number of animals reduces with time because of animal groups being perfused at the indicated time points. n = 3 (30, 90, and 120 min) or 6 (60 and 240 min) for each group of perfusion (stress; control only for 60 and 240 min). For IL-6 n = 3 or 6 (60 and 240 min) samples were analyzed *P < 0.05.
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Figure 1: Tb, motor activity, and plasma IL-6 levels during novel environment stress or control situation in rats. (A) Novel environment stress (stress) induced a rise in body temperature (Tb). (B) This response was preceded and accompanied by increased motor activity (averaged cumulative activity over 15 min for each) but (C) no significant rise in plasma interleukin 6 (IL-6)-levels, when compared over time to unstressed control animals. Dashed black arrows indicate changes in the number of animals; this number of animals reduces with time because of animal groups being perfused at the indicated time points. n = 3 (30, 90, and 120 min) or 6 (60 and 240 min) for each group of perfusion (stress; control only for 60 and 240 min). For IL-6 n = 3 or 6 (60 and 240 min) samples were analyzed *P < 0.05.

Mentions: Psychological stress induced an increase in Tb from 30 to 120 min (Figure 1A, P < 0.05; mean increase 1.03 ± 0.19 C; stress vs. control) and an increase in locomotor activity from 30 to 90 min (Figure 1B, P < 0.05), as previously reported [32]. Plasma IL-6 levels did not increase over time in the stressed animals (Figure 1C, one way ANOVA).


Activation of the inflammatory transcription factor nuclear factor interleukin-6 during inflammatory and psychological stress in the brain.

Fuchs F, Damm J, Gerstberger R, Roth J, Rummel C - J Neuroinflammation (2013)

Tb, motor activity, and plasma IL-6 levels during novel environment stress or control situation in rats. (A) Novel environment stress (stress) induced a rise in body temperature (Tb). (B) This response was preceded and accompanied by increased motor activity (averaged cumulative activity over 15 min for each) but (C) no significant rise in plasma interleukin 6 (IL-6)-levels, when compared over time to unstressed control animals. Dashed black arrows indicate changes in the number of animals; this number of animals reduces with time because of animal groups being perfused at the indicated time points. n = 3 (30, 90, and 120 min) or 6 (60 and 240 min) for each group of perfusion (stress; control only for 60 and 240 min). For IL-6 n = 3 or 6 (60 and 240 min) samples were analyzed *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4222273&req=5

Figure 1: Tb, motor activity, and plasma IL-6 levels during novel environment stress or control situation in rats. (A) Novel environment stress (stress) induced a rise in body temperature (Tb). (B) This response was preceded and accompanied by increased motor activity (averaged cumulative activity over 15 min for each) but (C) no significant rise in plasma interleukin 6 (IL-6)-levels, when compared over time to unstressed control animals. Dashed black arrows indicate changes in the number of animals; this number of animals reduces with time because of animal groups being perfused at the indicated time points. n = 3 (30, 90, and 120 min) or 6 (60 and 240 min) for each group of perfusion (stress; control only for 60 and 240 min). For IL-6 n = 3 or 6 (60 and 240 min) samples were analyzed *P < 0.05.
Mentions: Psychological stress induced an increase in Tb from 30 to 120 min (Figure 1A, P < 0.05; mean increase 1.03 ± 0.19 C; stress vs. control) and an increase in locomotor activity from 30 to 90 min (Figure 1B, P < 0.05), as previously reported [32]. Plasma IL-6 levels did not increase over time in the stressed animals (Figure 1C, one way ANOVA).

Bottom Line: In vitro NF-IL6-activation was associated with induction and secretion of TNFα in folliculostellate cells, which could be antagonized by the JAK-STAT-inhibitor AG490.Both stressors were associated with time-dependent activation of NF-IL6 in different cell types of the brain and the pituitary.Moreover, while NF-IL6-IR was partially linked to STAT3 and NFκB activation, TNFα production, and ACTH-IR after LPS stimulation; this was not the case after exposure to a novel environment, suggesting distinct underlying signaling pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Veterinary-Physiology and -Biochemistry, Justus-Liebig-University Giessen, Frankfurter Strasse 100, Giessen D-35392, Germany. Christoph.D.Rummel@vetmed.uni-giessen.de.

ABSTRACT

Background: The transcription factor nuclear factor interleukin 6 (NF-IL6) is known to be activated by various inflammatory stimuli in the brain. Interestingly, we recently detected NF-IL6-activation within the hypothalamus-pituitary-adrenal (HPA)-axis of rats after systemic lipopolysaccharide (LPS)-injection. Thus, the aim of the present study was to investigate whether NF-IL6 is activated during either, inflammatory, or psychological stress in the rat brain.

Methods: Rats were challenged with either the inflammatory stimulus LPS (100 μg/kg, i.p.) or exposed to a novel environment. Core body temperature (Tb) and motor activity were monitored using telemetry, animals were killed at different time points, brains and blood removed, and primary cell cultures of the anterior pituitary lobe (AL) were investigated. Analyses were performed using immunohistochemistry, RT-PCR, and cytokine-specific bioassays.

Results: Stress stimulation by a novel environment increased NF-IL6-immunoreactivity (IR) in the pituitary's perivascular macrophages and hypothalamic paraventricular cells and a rise in Tb lasting approximately 2 h. LPS stimulation lead to NF-IL6-IR in several additional cell types including ACTH-IR-positive corticotrope cells in vivo and in vitro. Two other proinflammatory transcription factors, namely signal transducer and activator of transcription (STAT)3 and NFκB, were significantly activated and partially colocalized with NF-IL6-IR in cells of the AL only after LPS-stimulation, but not following psychological stress. In vitro NF-IL6-activation was associated with induction and secretion of TNFα in folliculostellate cells, which could be antagonized by the JAK-STAT-inhibitor AG490.

Conclusions: We revealed, for the first time, that NF-IL6 activation occurs not only during inflammatory LPS stimulation, but also during psychological stress, that is, a novel environment. Both stressors were associated with time-dependent activation of NF-IL6 in different cell types of the brain and the pituitary. Moreover, while NF-IL6-IR was partially linked to STAT3 and NFκB activation, TNFα production, and ACTH-IR after LPS stimulation; this was not the case after exposure to a novel environment, suggesting distinct underlying signaling pathways. Overall, NF-IL6 can be used as a broad activation marker in the brain and might be of interest for therapeutic approaches not only during inflammatory but also psychological stress.

Show MeSH
Related in: MedlinePlus