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CD200R1 agonist attenuates glial activation, inflammatory reactions, and hypersensitivity immediately after its intrathecal application in a rat neuropathic pain model.

Hernangómez M, Klusáková I, Joukal M, Hradilová-Svíženská I, Guaza C, Dubový P - J Neuroinflammation (2016)

Bottom Line: Simultaneously, CD200Fc attenuated activation of glial cells and decreased proinflammatory and increased anti-inflammatory cytokine messenger RNA (mRNA) levels.The anti-inflammatory effect of CD200Fc dropped at 24 h after intrathecal application.Intrathecal administration of the CD200R1 agonist CD200Fc induces very rapid suppression of neuroinflammatory reactions associated with glial activation and neuropathic pain development.

View Article: PubMed Central - PubMed

Affiliation: Central European Institute of Technology (CEITEC), Masaryk University, Kamenice 3, 62500, Brno, Czech Republic. mirihh@hotmail.com.

ABSTRACT

Background: Interaction of CD200 with its receptor CD200R has an immunoregulatory role and attenuates various types of neuroinflammatory diseases.

Methods: Immunofluorescence staining, western blot analysis, and RT-PCR were used to investigate the modulatory effects of CD200 fusion protein (CD200Fc) on activation of microglia and astrocytes as well as synthesis of pro- (TNF, IL-1β, IL-6) and anti-inflammatory (IL-4, IL-10) cytokines in the L4-L5 spinal cord segments in relation to behavioral signs of neuropathic pain after unilateral sterile chronic constriction injury (sCCI) of the sciatic nerve. Withdrawal thresholds for mechanical hypersensitivity and latencies for thermal hypersensitivity were measured in hind paws 1 day before operation; 1, 3, and 7 days after sCCI operation; and then 5 and 24 h after intrathecal application of artificial cerebrospinal fluid or CD200Fc.

Results: Seven days from sCCI operation and 5 h from intrathecal application, CD200Fc reduced mechanical and thermal hypersensitivity when compared with control animals. Simultaneously, CD200Fc attenuated activation of glial cells and decreased proinflammatory and increased anti-inflammatory cytokine messenger RNA (mRNA) levels. Administration of CD200Fc also diminished elevation of CD200 and CD200R proteins as a concomitant reaction of the modulatory system to increased neuroinflammatory reactions after nerve injury. The anti-inflammatory effect of CD200Fc dropped at 24 h after intrathecal application.

Conclusions: Intrathecal administration of the CD200R1 agonist CD200Fc induces very rapid suppression of neuroinflammatory reactions associated with glial activation and neuropathic pain development. This may constitute a promising and novel therapeutic approach for the treatment of neuropathic pain.

No MeSH data available.


Related in: MedlinePlus

Double immunostaining to detect CD200 protein in astrocytes and neurons as well as CD200R in microglia. Double immunostaining of sections through DH of L4–L5 spinal cord segments from sCCI-operated and ACSF-treated rats. Immunofluorescence for CD200 (a) and GFAP (b) are colocalized (c), evidencing that CD200 protein is expressed by astrocytes (arrows). In addition, double immunostaining for CD200 (d) and NeuN (e) detected a presence of CD200 protein also in neurons (f, arrows). Immunolabeling for CD200R (g) and OX42 (h) was colocalized, illustrating that activated microglial cells display CD200R protein (i, arrow). Merged figures (c, f, i) contain also blue channel of Hoechst stained nuclei. Scale bars = 30 μm
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Fig5: Double immunostaining to detect CD200 protein in astrocytes and neurons as well as CD200R in microglia. Double immunostaining of sections through DH of L4–L5 spinal cord segments from sCCI-operated and ACSF-treated rats. Immunofluorescence for CD200 (a) and GFAP (b) are colocalized (c), evidencing that CD200 protein is expressed by astrocytes (arrows). In addition, double immunostaining for CD200 (d) and NeuN (e) detected a presence of CD200 protein also in neurons (f, arrows). Immunolabeling for CD200R (g) and OX42 (h) was colocalized, illustrating that activated microglial cells display CD200R protein (i, arrow). Merged figures (c, f, i) contain also blue channel of Hoechst stained nuclei. Scale bars = 30 μm

Mentions: Double immunostaining of spinal cord sections of sCCI-operated and ACSF-treated rats revealed CD200 immunoreaction in both NeuN+ neurons and GFAP+ astrocytes (Fig. 5a–f). The CD200R immunofluorescence was detected in neuropil of the spinal cord sections and double immunostaining displayed colocalization of CD200R and OX42 to illustrate presence of CD200R in activated microglial cells (Fig. 5g–i). When GFAP immunostaining corresponding with astrocyte activation was reduced after CD200Fc treatment, we sought to observe whether activated astrocytes could express CD200R1. Surprisingly, CD200R immunolabeling was colocalized with GFAP immunostaining, indicating expression of the receptor in activated astrocytes of the spinal cord from sCCI-operated rats (Fig. 6a–f). The unexpected colocalization was verified by image analysis and measurement of indexes using NIS Elements. Mander’s overlap measured in double immunostained cells indicated by arrows (Fig. 6d–f) was 0.965265, 0.929336, and 0.960253. In addition, profile intensity of red (CD200R) and green (GFAP) channels in the cells also indicated colocalization of CD200R and GFAP immunostaining (Fig. 6g).Fig. 5


CD200R1 agonist attenuates glial activation, inflammatory reactions, and hypersensitivity immediately after its intrathecal application in a rat neuropathic pain model.

Hernangómez M, Klusáková I, Joukal M, Hradilová-Svíženská I, Guaza C, Dubový P - J Neuroinflammation (2016)

Double immunostaining to detect CD200 protein in astrocytes and neurons as well as CD200R in microglia. Double immunostaining of sections through DH of L4–L5 spinal cord segments from sCCI-operated and ACSF-treated rats. Immunofluorescence for CD200 (a) and GFAP (b) are colocalized (c), evidencing that CD200 protein is expressed by astrocytes (arrows). In addition, double immunostaining for CD200 (d) and NeuN (e) detected a presence of CD200 protein also in neurons (f, arrows). Immunolabeling for CD200R (g) and OX42 (h) was colocalized, illustrating that activated microglial cells display CD200R protein (i, arrow). Merged figures (c, f, i) contain also blue channel of Hoechst stained nuclei. Scale bars = 30 μm
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4759712&req=5

Fig5: Double immunostaining to detect CD200 protein in astrocytes and neurons as well as CD200R in microglia. Double immunostaining of sections through DH of L4–L5 spinal cord segments from sCCI-operated and ACSF-treated rats. Immunofluorescence for CD200 (a) and GFAP (b) are colocalized (c), evidencing that CD200 protein is expressed by astrocytes (arrows). In addition, double immunostaining for CD200 (d) and NeuN (e) detected a presence of CD200 protein also in neurons (f, arrows). Immunolabeling for CD200R (g) and OX42 (h) was colocalized, illustrating that activated microglial cells display CD200R protein (i, arrow). Merged figures (c, f, i) contain also blue channel of Hoechst stained nuclei. Scale bars = 30 μm
Mentions: Double immunostaining of spinal cord sections of sCCI-operated and ACSF-treated rats revealed CD200 immunoreaction in both NeuN+ neurons and GFAP+ astrocytes (Fig. 5a–f). The CD200R immunofluorescence was detected in neuropil of the spinal cord sections and double immunostaining displayed colocalization of CD200R and OX42 to illustrate presence of CD200R in activated microglial cells (Fig. 5g–i). When GFAP immunostaining corresponding with astrocyte activation was reduced after CD200Fc treatment, we sought to observe whether activated astrocytes could express CD200R1. Surprisingly, CD200R immunolabeling was colocalized with GFAP immunostaining, indicating expression of the receptor in activated astrocytes of the spinal cord from sCCI-operated rats (Fig. 6a–f). The unexpected colocalization was verified by image analysis and measurement of indexes using NIS Elements. Mander’s overlap measured in double immunostained cells indicated by arrows (Fig. 6d–f) was 0.965265, 0.929336, and 0.960253. In addition, profile intensity of red (CD200R) and green (GFAP) channels in the cells also indicated colocalization of CD200R and GFAP immunostaining (Fig. 6g).Fig. 5

Bottom Line: Simultaneously, CD200Fc attenuated activation of glial cells and decreased proinflammatory and increased anti-inflammatory cytokine messenger RNA (mRNA) levels.The anti-inflammatory effect of CD200Fc dropped at 24 h after intrathecal application.Intrathecal administration of the CD200R1 agonist CD200Fc induces very rapid suppression of neuroinflammatory reactions associated with glial activation and neuropathic pain development.

View Article: PubMed Central - PubMed

Affiliation: Central European Institute of Technology (CEITEC), Masaryk University, Kamenice 3, 62500, Brno, Czech Republic. mirihh@hotmail.com.

ABSTRACT

Background: Interaction of CD200 with its receptor CD200R has an immunoregulatory role and attenuates various types of neuroinflammatory diseases.

Methods: Immunofluorescence staining, western blot analysis, and RT-PCR were used to investigate the modulatory effects of CD200 fusion protein (CD200Fc) on activation of microglia and astrocytes as well as synthesis of pro- (TNF, IL-1β, IL-6) and anti-inflammatory (IL-4, IL-10) cytokines in the L4-L5 spinal cord segments in relation to behavioral signs of neuropathic pain after unilateral sterile chronic constriction injury (sCCI) of the sciatic nerve. Withdrawal thresholds for mechanical hypersensitivity and latencies for thermal hypersensitivity were measured in hind paws 1 day before operation; 1, 3, and 7 days after sCCI operation; and then 5 and 24 h after intrathecal application of artificial cerebrospinal fluid or CD200Fc.

Results: Seven days from sCCI operation and 5 h from intrathecal application, CD200Fc reduced mechanical and thermal hypersensitivity when compared with control animals. Simultaneously, CD200Fc attenuated activation of glial cells and decreased proinflammatory and increased anti-inflammatory cytokine messenger RNA (mRNA) levels. Administration of CD200Fc also diminished elevation of CD200 and CD200R proteins as a concomitant reaction of the modulatory system to increased neuroinflammatory reactions after nerve injury. The anti-inflammatory effect of CD200Fc dropped at 24 h after intrathecal application.

Conclusions: Intrathecal administration of the CD200R1 agonist CD200Fc induces very rapid suppression of neuroinflammatory reactions associated with glial activation and neuropathic pain development. This may constitute a promising and novel therapeutic approach for the treatment of neuropathic pain.

No MeSH data available.


Related in: MedlinePlus