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An anti-inflammatory role for C/EBPδ in human brain pericytes.

Rustenhoven J, Scotter EL, Jansson D, Kho DT, Oldfield RL, Bergin PS, Mee EW, Faull RL, Curtis MA, Graham SE, Park TI, Dragunow M - Sci Rep (2015)

Bottom Line: Our results reveal that C/EBPδ is induced in a concentration- and time-dependent fashion in human brain pericytes by interleukin-1β (IL-1β).C/EBPδ knockdown enhanced IL-1β-induced production of intracellular adhesion molecule-1 (ICAM-1), interleukin-8, monocyte chemoattractant protein-1 (MCP-1) and IL-1β, whilst attenuating cyclooxygenase-2 and superoxide dismutase-2 gene expression.Altered ICAM-1 and MCP-1 protein expression were confirmed by cytometric bead array and immunocytochemistry.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Pharmacology and Clinical Pharmacology [2] Centre for Brain Research.

ABSTRACT
Neuroinflammation contributes to the pathogenesis of several neurological disorders and pericytes are implicated in brain inflammatory processes. Cellular inflammatory responses are orchestrated by transcription factors but information on transcriptional control in pericytes is lacking. Because the transcription factor CCAAT/enhancer binding protein delta (C/EBPδ) is induced in a number of inflammatory brain disorders, we sought to investigate its role in regulating pericyte immune responses. Our results reveal that C/EBPδ is induced in a concentration- and time-dependent fashion in human brain pericytes by interleukin-1β (IL-1β). To investigate the function of the induced C/EBPδ in pericytes we used siRNA to knockdown IL-1β-induced C/EBPδ expression. C/EBPδ knockdown enhanced IL-1β-induced production of intracellular adhesion molecule-1 (ICAM-1), interleukin-8, monocyte chemoattractant protein-1 (MCP-1) and IL-1β, whilst attenuating cyclooxygenase-2 and superoxide dismutase-2 gene expression. Altered ICAM-1 and MCP-1 protein expression were confirmed by cytometric bead array and immunocytochemistry. Our results show that knock-down of C/EBPδ expression in pericytes following immune stimulation increased chemokine and adhesion molecule expression, thus modifying the human brain pericyte inflammatory response. The induction of C/EBPδ following immune stimulation may act to limit infiltration of peripheral immune cells, thereby preventing further inflammatory responses in the brain.

No MeSH data available.


Related in: MedlinePlus

C/EBPδ is induced in human brain pericytes by IL-1β/IFNγ.Human brain pericytes were treated with vehicle or 10 ng/mL IL-1β + IFNγ for 24 hours and RNA was extracted. Expression of C/EBPα, C/EBPβ and C/EBPδ was determined by microarray10 (a) and qRT-PCR (b). Data is displayed as average fold change of five independent cases (a) or mean ± SEM of a separate three independent cases (b). *** = p < 0.001.
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f2: C/EBPδ is induced in human brain pericytes by IL-1β/IFNγ.Human brain pericytes were treated with vehicle or 10 ng/mL IL-1β + IFNγ for 24 hours and RNA was extracted. Expression of C/EBPα, C/EBPβ and C/EBPδ was determined by microarray10 (a) and qRT-PCR (b). Data is displayed as average fold change of five independent cases (a) or mean ± SEM of a separate three independent cases (b). *** = p < 0.001.

Mentions: Microarray analyis of human brain pericytes treated with IL-1β/IFNγ has previously revealed an induction of numerous inflammatory genes10. Due to the ability of the C/EBP family of transcription factors to modify cellular inflammatory responses, we chose to investigate the induction of three members C/EBPα, C/EBPβ and C/EBPδ in this dataset. By microarray analysis, C/EBPδ was found to be significantly increased by an IL-1β/IFNγ treatment (p < 0.001) whilst C/EBPα (p > 0.05) and C/EBPβ (p < 0.05) were not affected (Fig. 2a). Real time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) was performed on independent RNA samples, which confirmed the induction of C/EBPδ (5.33 ± 0.69 fold; p < 0.001), whereas no significant change in C/EBPα (0.74 ± 0.18 fold; p > 0.05) or C/EBPβ (1.18 ± 0.15 fold; p > 0.05) was observed (Fig. 2b), consistent with the microarray data.


An anti-inflammatory role for C/EBPδ in human brain pericytes.

Rustenhoven J, Scotter EL, Jansson D, Kho DT, Oldfield RL, Bergin PS, Mee EW, Faull RL, Curtis MA, Graham SE, Park TI, Dragunow M - Sci Rep (2015)

C/EBPδ is induced in human brain pericytes by IL-1β/IFNγ.Human brain pericytes were treated with vehicle or 10 ng/mL IL-1β + IFNγ for 24 hours and RNA was extracted. Expression of C/EBPα, C/EBPβ and C/EBPδ was determined by microarray10 (a) and qRT-PCR (b). Data is displayed as average fold change of five independent cases (a) or mean ± SEM of a separate three independent cases (b). *** = p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4499812&req=5

f2: C/EBPδ is induced in human brain pericytes by IL-1β/IFNγ.Human brain pericytes were treated with vehicle or 10 ng/mL IL-1β + IFNγ for 24 hours and RNA was extracted. Expression of C/EBPα, C/EBPβ and C/EBPδ was determined by microarray10 (a) and qRT-PCR (b). Data is displayed as average fold change of five independent cases (a) or mean ± SEM of a separate three independent cases (b). *** = p < 0.001.
Mentions: Microarray analyis of human brain pericytes treated with IL-1β/IFNγ has previously revealed an induction of numerous inflammatory genes10. Due to the ability of the C/EBP family of transcription factors to modify cellular inflammatory responses, we chose to investigate the induction of three members C/EBPα, C/EBPβ and C/EBPδ in this dataset. By microarray analysis, C/EBPδ was found to be significantly increased by an IL-1β/IFNγ treatment (p < 0.001) whilst C/EBPα (p > 0.05) and C/EBPβ (p < 0.05) were not affected (Fig. 2a). Real time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) was performed on independent RNA samples, which confirmed the induction of C/EBPδ (5.33 ± 0.69 fold; p < 0.001), whereas no significant change in C/EBPα (0.74 ± 0.18 fold; p > 0.05) or C/EBPβ (1.18 ± 0.15 fold; p > 0.05) was observed (Fig. 2b), consistent with the microarray data.

Bottom Line: Our results reveal that C/EBPδ is induced in a concentration- and time-dependent fashion in human brain pericytes by interleukin-1β (IL-1β).C/EBPδ knockdown enhanced IL-1β-induced production of intracellular adhesion molecule-1 (ICAM-1), interleukin-8, monocyte chemoattractant protein-1 (MCP-1) and IL-1β, whilst attenuating cyclooxygenase-2 and superoxide dismutase-2 gene expression.Altered ICAM-1 and MCP-1 protein expression were confirmed by cytometric bead array and immunocytochemistry.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Pharmacology and Clinical Pharmacology [2] Centre for Brain Research.

ABSTRACT
Neuroinflammation contributes to the pathogenesis of several neurological disorders and pericytes are implicated in brain inflammatory processes. Cellular inflammatory responses are orchestrated by transcription factors but information on transcriptional control in pericytes is lacking. Because the transcription factor CCAAT/enhancer binding protein delta (C/EBPδ) is induced in a number of inflammatory brain disorders, we sought to investigate its role in regulating pericyte immune responses. Our results reveal that C/EBPδ is induced in a concentration- and time-dependent fashion in human brain pericytes by interleukin-1β (IL-1β). To investigate the function of the induced C/EBPδ in pericytes we used siRNA to knockdown IL-1β-induced C/EBPδ expression. C/EBPδ knockdown enhanced IL-1β-induced production of intracellular adhesion molecule-1 (ICAM-1), interleukin-8, monocyte chemoattractant protein-1 (MCP-1) and IL-1β, whilst attenuating cyclooxygenase-2 and superoxide dismutase-2 gene expression. Altered ICAM-1 and MCP-1 protein expression were confirmed by cytometric bead array and immunocytochemistry. Our results show that knock-down of C/EBPδ expression in pericytes following immune stimulation increased chemokine and adhesion molecule expression, thus modifying the human brain pericyte inflammatory response. The induction of C/EBPδ following immune stimulation may act to limit infiltration of peripheral immune cells, thereby preventing further inflammatory responses in the brain.

No MeSH data available.


Related in: MedlinePlus