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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 knockdown modifies IL-1β induced inflammatory gene expression.Human brain pericytes were transfected with 50 nM of control or C/EBPδ siRNA for 48 hours. Following transfection, cells were treated with vehicle or 10 ng/mL IL-1β for six hours and RNA was extracted. The effect of C/EBPδ knockdown on IL-1β (a), ICAM-1 (b), MCP-1 (c), IL-8(d), SOD2 (e), COX-2 (f) and IL-6 (g) was assayed by qRT-PCR. Data is displayed as mean ± SEM from four independent cases ** = p < 0.01, *** = p < 0.001.
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f6: C/EBPδ knockdown modifies IL-1β induced inflammatory gene expression.Human brain pericytes were transfected with 50 nM of control or C/EBPδ siRNA for 48 hours. Following transfection, cells were treated with vehicle or 10 ng/mL IL-1β for six hours and RNA was extracted. The effect of C/EBPδ knockdown on IL-1β (a), ICAM-1 (b), MCP-1 (c), IL-8(d), SOD2 (e), COX-2 (f) and IL-6 (g) was assayed by qRT-PCR. Data is displayed as mean ± SEM from four independent cases ** = p < 0.01, *** = p < 0.001.

Mentions: Having observed a reduction in IL-1β induced C/EBPδ expression with siRNA transfection, we examined the effect on a range of inflammatory mediators which have previously been shown to be induced by brain pericytes with inflammatory stimuli. C/EBPδ knockdown resulted in increased IL-1β-induced expression of IL-1β (control siRNA 29.74 ± 4.13 fold, C/EBPδ siRNA 55.88 ± 4.99 fold; p < 0.001; Fig. 6a), ICAM-1 (control siRNA 125.20 ± 5.40 fold, C/EBPδ siRNA 295.72 ± 41.51 fold; p < 0.001; Fig. 6b), MCP-1 (control siRNA 19.04 ± 1.39 fold, C/EBPδ siRNA 32.59 ± 1.61 fold; p < 0.001; Fig. 6c) and IL-8 (control siRNA 634.81 ± 38.01 fold, C/EBPδ siRNA 1021.12 ± 121.10 fold; p<0.001; Fig. 6d). In contrast, attenuated expression was observed for SOD2 (control siRNA 56.72 ± 4.47 fold, C/EBPδ siRNA 41.50 ± 1.98 fold; p < 0.01; Fig. 6e) and COX-2 (control siRNA 6.35 ± 4.86 fold, C/EBPδ siRNA 4.50 ± 1.14 fold; p < 0.001; Fig. 6f), whilst a non-significant decrease was observed for IL-6 (control siRNA 530.91 ± 174.61 fold, C/EBPδ siRNA 369.92 ± 133.60 fold; p > 0.05; Fig. 6g). The basal expression of all inflammatory genes showed no change with C/EBPδ or control siRNA.


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 knockdown modifies IL-1β induced inflammatory gene expression.Human brain pericytes were transfected with 50 nM of control or C/EBPδ siRNA for 48 hours. Following transfection, cells were treated with vehicle or 10 ng/mL IL-1β for six hours and RNA was extracted. The effect of C/EBPδ knockdown on IL-1β (a), ICAM-1 (b), MCP-1 (c), IL-8(d), SOD2 (e), COX-2 (f) and IL-6 (g) was assayed by qRT-PCR. Data is displayed as mean ± SEM from four independent cases ** = p < 0.01, *** = p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f6: C/EBPδ knockdown modifies IL-1β induced inflammatory gene expression.Human brain pericytes were transfected with 50 nM of control or C/EBPδ siRNA for 48 hours. Following transfection, cells were treated with vehicle or 10 ng/mL IL-1β for six hours and RNA was extracted. The effect of C/EBPδ knockdown on IL-1β (a), ICAM-1 (b), MCP-1 (c), IL-8(d), SOD2 (e), COX-2 (f) and IL-6 (g) was assayed by qRT-PCR. Data is displayed as mean ± SEM from four independent cases ** = p < 0.01, *** = p < 0.001.
Mentions: Having observed a reduction in IL-1β induced C/EBPδ expression with siRNA transfection, we examined the effect on a range of inflammatory mediators which have previously been shown to be induced by brain pericytes with inflammatory stimuli. C/EBPδ knockdown resulted in increased IL-1β-induced expression of IL-1β (control siRNA 29.74 ± 4.13 fold, C/EBPδ siRNA 55.88 ± 4.99 fold; p < 0.001; Fig. 6a), ICAM-1 (control siRNA 125.20 ± 5.40 fold, C/EBPδ siRNA 295.72 ± 41.51 fold; p < 0.001; Fig. 6b), MCP-1 (control siRNA 19.04 ± 1.39 fold, C/EBPδ siRNA 32.59 ± 1.61 fold; p < 0.001; Fig. 6c) and IL-8 (control siRNA 634.81 ± 38.01 fold, C/EBPδ siRNA 1021.12 ± 121.10 fold; p<0.001; Fig. 6d). In contrast, attenuated expression was observed for SOD2 (control siRNA 56.72 ± 4.47 fold, C/EBPδ siRNA 41.50 ± 1.98 fold; p < 0.01; Fig. 6e) and COX-2 (control siRNA 6.35 ± 4.86 fold, C/EBPδ siRNA 4.50 ± 1.14 fold; p < 0.001; Fig. 6f), whilst a non-significant decrease was observed for IL-6 (control siRNA 530.91 ± 174.61 fold, C/EBPδ siRNA 369.92 ± 133.60 fold; p > 0.05; Fig. 6g). The basal expression of all inflammatory genes showed no change with C/EBPδ or control siRNA.

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