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Repetitive hyperbaric oxygenation attenuates reactive astrogliosis and suppresses expression of inflammatory mediators in the rat model of brain injury.

Lavrnja I, Parabucki A, Brkic P, Jovanovic T, Dacic S, Savic D, Pantic I, Stojiljkovic M, Pekovic S - Mediators Inflamm. (2015)

Bottom Line: The exact mechanisms by which treatment with hyperbaric oxygen (HBOT) exerts its beneficial effects on recovery after brain injury are still unrevealed.Data obtained using real-time polymerase chain reaction, Western blot, and immunohistochemical and immunofluorescence analyses revealed that repetitive HBOT applied after the CSI attenuates reactive astrogliosis and glial scarring, and reduces expression of GFAP (glial fibrillary acidic protein), vimentin, and ICAM-1 (intercellular adhesion molecule-1) both at gene and tissue levels.Accordingly, repetitive HBOT, by prevention of glial scarring and limiting of expression of inflammatory mediators, supports formation of more permissive environment for repair and regeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology, Institute for Biological Research "Sinisa Stankovic", University of Belgrade, 11060 Belgrade, Serbia.

ABSTRACT
The exact mechanisms by which treatment with hyperbaric oxygen (HBOT) exerts its beneficial effects on recovery after brain injury are still unrevealed. Therefore, in this study we investigated the influence of repetitive HBOT on the reactive astrogliosis and expression of mediators of inflammation after cortical stab injury (CSI). CSI was performed on male Wistar rats, divided into control, sham, and lesioned groups with appropriate HBO. The HBOT protocol was as follows: 10 minutes of slow compression, 2.5 atmospheres absolute (ATA) for 60 minutes, and 10 minutes of slow decompression, once a day for 10 consecutive days. Data obtained using real-time polymerase chain reaction, Western blot, and immunohistochemical and immunofluorescence analyses revealed that repetitive HBOT applied after the CSI attenuates reactive astrogliosis and glial scarring, and reduces expression of GFAP (glial fibrillary acidic protein), vimentin, and ICAM-1 (intercellular adhesion molecule-1) both at gene and tissue levels. In addition, HBOT prevents expression of CD40 and its ligand CD40L on microglia, neutrophils, cortical neurons, and reactive astrocytes. Accordingly, repetitive HBOT, by prevention of glial scarring and limiting of expression of inflammatory mediators, supports formation of more permissive environment for repair and regeneration.

No MeSH data available.


Related in: MedlinePlus

Double-immunofluorescence analysis of CD40L (green) and GFAP (red) colocalization after CSI and HBOT. ((a)–(c)) Sparse CD40L/GFAP positive fibrous astrocytes were seen in the cortex of control rats. ((d)–(f)) Strong CD40L immunoreactivity occurred in GFAP+ astrocytes that form dense mesh of glial scar in the area adjacent to the lesion site, providing an overlapping signal (yellow fluorescence). ((g)–(i)) CD40L and GFAP signal is abundantly present at protoplasmic astrocyte cell bodies, and thick proximal and distal processes. ((j)–(l)) Repetitive HBOT decreased intensity of CD40L/GFAP immunofluorescence. ((m)–(o)) Reactive phenotype of astrocytes is transformed into more resting form with smaller cell body and long processes resembling morphology of astrocytes from the control group. Scale bar = 50 μm.
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fig8: Double-immunofluorescence analysis of CD40L (green) and GFAP (red) colocalization after CSI and HBOT. ((a)–(c)) Sparse CD40L/GFAP positive fibrous astrocytes were seen in the cortex of control rats. ((d)–(f)) Strong CD40L immunoreactivity occurred in GFAP+ astrocytes that form dense mesh of glial scar in the area adjacent to the lesion site, providing an overlapping signal (yellow fluorescence). ((g)–(i)) CD40L and GFAP signal is abundantly present at protoplasmic astrocyte cell bodies, and thick proximal and distal processes. ((j)–(l)) Repetitive HBOT decreased intensity of CD40L/GFAP immunofluorescence. ((m)–(o)) Reactive phenotype of astrocytes is transformed into more resting form with smaller cell body and long processes resembling morphology of astrocytes from the control group. Scale bar = 50 μm.

Mentions: In order to better characterize astrocytic expression of CD40L after CSI and HBOT we performed double-immunofluorescence staining (Figure 8). The CD40L-labeled astrocytes were identified with the antibody against CD40L (green fluorescence) and the astrocytic marker GFAP (red fluorescence). No apparent differences in the pattern of CD40L/GFAP immunostaining between all controls (C, CHBO, S, and SHBO) were noted (data shown as supplemented material). Populations of CD40L/GFAP positive fibrous astrocytes with small cell bodies and thin processes were scattered throughout the cortex of control rats (Figure 8(c)). After the injury, strong CD40L immunoreactivity occurred in GFAP+ astrocytes clustered around the lesion site forming dense mesh of glial scar composed of tightly interweaved cell processes. CD40L immunofluorescence completely overlaps with GFAP resulting in yellow fluorescence (Figure 8(f)). This strong CD40L and GFAP immunoreactivity occurred in the hypertrophied cell body, as well as in the thick proximal and distal processes (Figure 8(i)). After repetitive HBOT, reactive phenotype of astrocytes is changed into resting form with smaller cell body and long processes. CD40L is expressed only in subpopulation of fibrous astrocytes around the lesion site (Figures 8(j)–8(l)). The morphology of these astrocytes was similar to astrocytes from control group (Figures 8(m)–8(o)).


Repetitive hyperbaric oxygenation attenuates reactive astrogliosis and suppresses expression of inflammatory mediators in the rat model of brain injury.

Lavrnja I, Parabucki A, Brkic P, Jovanovic T, Dacic S, Savic D, Pantic I, Stojiljkovic M, Pekovic S - Mediators Inflamm. (2015)

Double-immunofluorescence analysis of CD40L (green) and GFAP (red) colocalization after CSI and HBOT. ((a)–(c)) Sparse CD40L/GFAP positive fibrous astrocytes were seen in the cortex of control rats. ((d)–(f)) Strong CD40L immunoreactivity occurred in GFAP+ astrocytes that form dense mesh of glial scar in the area adjacent to the lesion site, providing an overlapping signal (yellow fluorescence). ((g)–(i)) CD40L and GFAP signal is abundantly present at protoplasmic astrocyte cell bodies, and thick proximal and distal processes. ((j)–(l)) Repetitive HBOT decreased intensity of CD40L/GFAP immunofluorescence. ((m)–(o)) Reactive phenotype of astrocytes is transformed into more resting form with smaller cell body and long processes resembling morphology of astrocytes from the control group. Scale bar = 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4417949&req=5

fig8: Double-immunofluorescence analysis of CD40L (green) and GFAP (red) colocalization after CSI and HBOT. ((a)–(c)) Sparse CD40L/GFAP positive fibrous astrocytes were seen in the cortex of control rats. ((d)–(f)) Strong CD40L immunoreactivity occurred in GFAP+ astrocytes that form dense mesh of glial scar in the area adjacent to the lesion site, providing an overlapping signal (yellow fluorescence). ((g)–(i)) CD40L and GFAP signal is abundantly present at protoplasmic astrocyte cell bodies, and thick proximal and distal processes. ((j)–(l)) Repetitive HBOT decreased intensity of CD40L/GFAP immunofluorescence. ((m)–(o)) Reactive phenotype of astrocytes is transformed into more resting form with smaller cell body and long processes resembling morphology of astrocytes from the control group. Scale bar = 50 μm.
Mentions: In order to better characterize astrocytic expression of CD40L after CSI and HBOT we performed double-immunofluorescence staining (Figure 8). The CD40L-labeled astrocytes were identified with the antibody against CD40L (green fluorescence) and the astrocytic marker GFAP (red fluorescence). No apparent differences in the pattern of CD40L/GFAP immunostaining between all controls (C, CHBO, S, and SHBO) were noted (data shown as supplemented material). Populations of CD40L/GFAP positive fibrous astrocytes with small cell bodies and thin processes were scattered throughout the cortex of control rats (Figure 8(c)). After the injury, strong CD40L immunoreactivity occurred in GFAP+ astrocytes clustered around the lesion site forming dense mesh of glial scar composed of tightly interweaved cell processes. CD40L immunofluorescence completely overlaps with GFAP resulting in yellow fluorescence (Figure 8(f)). This strong CD40L and GFAP immunoreactivity occurred in the hypertrophied cell body, as well as in the thick proximal and distal processes (Figure 8(i)). After repetitive HBOT, reactive phenotype of astrocytes is changed into resting form with smaller cell body and long processes. CD40L is expressed only in subpopulation of fibrous astrocytes around the lesion site (Figures 8(j)–8(l)). The morphology of these astrocytes was similar to astrocytes from control group (Figures 8(m)–8(o)).

Bottom Line: The exact mechanisms by which treatment with hyperbaric oxygen (HBOT) exerts its beneficial effects on recovery after brain injury are still unrevealed.Data obtained using real-time polymerase chain reaction, Western blot, and immunohistochemical and immunofluorescence analyses revealed that repetitive HBOT applied after the CSI attenuates reactive astrogliosis and glial scarring, and reduces expression of GFAP (glial fibrillary acidic protein), vimentin, and ICAM-1 (intercellular adhesion molecule-1) both at gene and tissue levels.Accordingly, repetitive HBOT, by prevention of glial scarring and limiting of expression of inflammatory mediators, supports formation of more permissive environment for repair and regeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology, Institute for Biological Research "Sinisa Stankovic", University of Belgrade, 11060 Belgrade, Serbia.

ABSTRACT
The exact mechanisms by which treatment with hyperbaric oxygen (HBOT) exerts its beneficial effects on recovery after brain injury are still unrevealed. Therefore, in this study we investigated the influence of repetitive HBOT on the reactive astrogliosis and expression of mediators of inflammation after cortical stab injury (CSI). CSI was performed on male Wistar rats, divided into control, sham, and lesioned groups with appropriate HBO. The HBOT protocol was as follows: 10 minutes of slow compression, 2.5 atmospheres absolute (ATA) for 60 minutes, and 10 minutes of slow decompression, once a day for 10 consecutive days. Data obtained using real-time polymerase chain reaction, Western blot, and immunohistochemical and immunofluorescence analyses revealed that repetitive HBOT applied after the CSI attenuates reactive astrogliosis and glial scarring, and reduces expression of GFAP (glial fibrillary acidic protein), vimentin, and ICAM-1 (intercellular adhesion molecule-1) both at gene and tissue levels. In addition, HBOT prevents expression of CD40 and its ligand CD40L on microglia, neutrophils, cortical neurons, and reactive astrocytes. Accordingly, repetitive HBOT, by prevention of glial scarring and limiting of expression of inflammatory mediators, supports formation of more permissive environment for repair and regeneration.

No MeSH data available.


Related in: MedlinePlus