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Mesenchymal stem cells stabilize the blood-brain barrier through regulation of astrocytes.

Park HJ, Shin JY, Kim HN, Oh SH, Song SK, Lee PH - Stem Cell Res Ther (2015)

Bottom Line: Consequently, MSC treatment reduced neutrophil infiltration and enhanced survival of midbrain dopaminergic neurons in LPS-treated animals.In cellular system, MSC treatment led to a significant reversion of VEGF-A-induced eNOS and tight junction protein expression in endothelial cells, which led to increased EBA expressing cells.Additionally, MSC treatment significantly attenuated LPS-induced increased expressions of IL-1β in microglia and VEGF-A in astrocytes with an increase in IL-10 levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology and Brain Research Institute, Yonsei University College of Medicine, 250 Seongsanno, Seodaemun-gu, Seoul, 120-752, South Korea. pphj0105@yuhs.ac.

ABSTRACT

Introduction: The blood-brain barrier (BBB) protects the brain against potentially neurotoxic molecules in the circulation, and loss of its integrity may contribute to disease progression in neurodegenerative conditions. Recently, the active role of reactive astrocytes in BBB disruption has become evident in the inflamed brain. In the present study, we investigated whether mesenchymal stem cell (MSC) treatment might modulate reactive astrocytes and thus stabilize BBB integrity through vascular endothelial growth factor A (VEGF-A) signaling in inflammatory conditions.

Methods: For the inflamed brain, we injected LPS using a stereotaxic apparatus and MSCs were injected into the tail vein. At 6 hours and 7 days after LPS injection, we analyzed modulatory effects of MSCs on the change of BBB permeability through VEGF-A signaling using immunochemistry and western blot. To determine the effects of MSCs on VEGF-A-related signaling in cellular system, we had used endothelial cells treated with VEGF-A and co-cultured astrocyte and BV 2 cells treated with lipopolysaccharide (LPS) and then these cells were co-cultured with MSCs.

Results: In LPS-treated rats, MSCs restored Evans blue infiltration and the number of endothelial-barrier antigen (EBA) and P-glycoprotein (p-gp)-expressing cells, which were significantly altered in LPS-treated animals. Additionally, MSC administration following LPS treatment markedly increased the density of astrocytic filaments around vessels and reversed LPS-induced elevations in VEGF-A levels as well as endothelial nitric oxide synthase (eNOS)-dependent downregulation of tight junction proteins in the endothelium. Consequently, MSC treatment reduced neutrophil infiltration and enhanced survival of midbrain dopaminergic neurons in LPS-treated animals. In cellular system, MSC treatment led to a significant reversion of VEGF-A-induced eNOS and tight junction protein expression in endothelial cells, which led to increased EBA expressing cells. Additionally, MSC treatment significantly attenuated LPS-induced increased expressions of IL-1β in microglia and VEGF-A in astrocytes with an increase in IL-10 levels.

Conclusion: The present study indicated that MSCs may stabilize BBB permeability by modulating astrocytic endfeet and VEGF-A signaling, which may be relevant to the treatment of Parkinsonian diseases as a candidate for disease modifying therapeutics.

No MeSH data available.


Related in: MedlinePlus

MSC treatment reduces LPS-induced neutrophil infiltration and loss of dopaminergic cells in the midbrain. There was a marked increase in MPO-positive cells in LPS-induced animals; however, MSC treatment following LPS administration notably decreased the number of these cells a (n = 5). **p <0.001. Scale bar: 100 μm. Consequently, MSC treatment inhibited LPS-induced loss of TH-positive cells in the midbrain b (n = 5). *p <0.05. Behavioral analysis showed that the number of rotations after apomorphine injection was significantly increased in animals receiving LPS compared with controls, whereas MSC administration led to a significant reduction of rotation behavior in LPS-treated animals c (n = 5). **p <0.001. Scale bar: 100 μm. LPS, lipopolysaccharide, MSC mesenchymal stem cell, MPO myeloperoxidase, TH tyrosine hydroxylase
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Fig4: MSC treatment reduces LPS-induced neutrophil infiltration and loss of dopaminergic cells in the midbrain. There was a marked increase in MPO-positive cells in LPS-induced animals; however, MSC treatment following LPS administration notably decreased the number of these cells a (n = 5). **p <0.001. Scale bar: 100 μm. Consequently, MSC treatment inhibited LPS-induced loss of TH-positive cells in the midbrain b (n = 5). *p <0.05. Behavioral analysis showed that the number of rotations after apomorphine injection was significantly increased in animals receiving LPS compared with controls, whereas MSC administration led to a significant reduction of rotation behavior in LPS-treated animals c (n = 5). **p <0.001. Scale bar: 100 μm. LPS, lipopolysaccharide, MSC mesenchymal stem cell, MPO myeloperoxidase, TH tyrosine hydroxylase

Mentions: To determine the effects of MSCs on neutrophil infiltration, SN tissue was immunostained for MPO, a marker for activated neutrophils. There was a marked increase in MPO-positive cells in LPS-induced animals; however, MSC treatment following LPS administration notably decreased these cells (Fig. 4a). Stereological analysis revealed that the number of neutrophils in the SN was significantly decreased in MSC-treated rats relative to those treated only with LPS (p <0.001; Fig. 4a). Consequently, MSC treatment decreased LPS-induced loss of dopaminergic neurons (Fig. 4b). On stereological analysis, the number of TH-positive cells in the SN was significantly increased in MSC-treated rats relative to that in rats treated only with LPS (p <0.05; Fig. 4b). Behavioral analysis showed that the number of rotations after apomorphine injection was significantly increased in animals receiving LPS compared with controls. However, MSC administration in LPS-treated animals led to a significant reduction of rotation behavior at 1 day and 7 days after LPS injection (Fig. 4c). We attempted to identify MSCs in the MSC-treated animals using human-specific NuMA immunostaining. The histological analysis showed that NuMA-positive cells were observed in the left SN (see Additional file 1).Fig. 4


Mesenchymal stem cells stabilize the blood-brain barrier through regulation of astrocytes.

Park HJ, Shin JY, Kim HN, Oh SH, Song SK, Lee PH - Stem Cell Res Ther (2015)

MSC treatment reduces LPS-induced neutrophil infiltration and loss of dopaminergic cells in the midbrain. There was a marked increase in MPO-positive cells in LPS-induced animals; however, MSC treatment following LPS administration notably decreased the number of these cells a (n = 5). **p <0.001. Scale bar: 100 μm. Consequently, MSC treatment inhibited LPS-induced loss of TH-positive cells in the midbrain b (n = 5). *p <0.05. Behavioral analysis showed that the number of rotations after apomorphine injection was significantly increased in animals receiving LPS compared with controls, whereas MSC administration led to a significant reduction of rotation behavior in LPS-treated animals c (n = 5). **p <0.001. Scale bar: 100 μm. LPS, lipopolysaccharide, MSC mesenchymal stem cell, MPO myeloperoxidase, TH tyrosine hydroxylase
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4588687&req=5

Fig4: MSC treatment reduces LPS-induced neutrophil infiltration and loss of dopaminergic cells in the midbrain. There was a marked increase in MPO-positive cells in LPS-induced animals; however, MSC treatment following LPS administration notably decreased the number of these cells a (n = 5). **p <0.001. Scale bar: 100 μm. Consequently, MSC treatment inhibited LPS-induced loss of TH-positive cells in the midbrain b (n = 5). *p <0.05. Behavioral analysis showed that the number of rotations after apomorphine injection was significantly increased in animals receiving LPS compared with controls, whereas MSC administration led to a significant reduction of rotation behavior in LPS-treated animals c (n = 5). **p <0.001. Scale bar: 100 μm. LPS, lipopolysaccharide, MSC mesenchymal stem cell, MPO myeloperoxidase, TH tyrosine hydroxylase
Mentions: To determine the effects of MSCs on neutrophil infiltration, SN tissue was immunostained for MPO, a marker for activated neutrophils. There was a marked increase in MPO-positive cells in LPS-induced animals; however, MSC treatment following LPS administration notably decreased these cells (Fig. 4a). Stereological analysis revealed that the number of neutrophils in the SN was significantly decreased in MSC-treated rats relative to those treated only with LPS (p <0.001; Fig. 4a). Consequently, MSC treatment decreased LPS-induced loss of dopaminergic neurons (Fig. 4b). On stereological analysis, the number of TH-positive cells in the SN was significantly increased in MSC-treated rats relative to that in rats treated only with LPS (p <0.05; Fig. 4b). Behavioral analysis showed that the number of rotations after apomorphine injection was significantly increased in animals receiving LPS compared with controls. However, MSC administration in LPS-treated animals led to a significant reduction of rotation behavior at 1 day and 7 days after LPS injection (Fig. 4c). We attempted to identify MSCs in the MSC-treated animals using human-specific NuMA immunostaining. The histological analysis showed that NuMA-positive cells were observed in the left SN (see Additional file 1).Fig. 4

Bottom Line: Consequently, MSC treatment reduced neutrophil infiltration and enhanced survival of midbrain dopaminergic neurons in LPS-treated animals.In cellular system, MSC treatment led to a significant reversion of VEGF-A-induced eNOS and tight junction protein expression in endothelial cells, which led to increased EBA expressing cells.Additionally, MSC treatment significantly attenuated LPS-induced increased expressions of IL-1β in microglia and VEGF-A in astrocytes with an increase in IL-10 levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology and Brain Research Institute, Yonsei University College of Medicine, 250 Seongsanno, Seodaemun-gu, Seoul, 120-752, South Korea. pphj0105@yuhs.ac.

ABSTRACT

Introduction: The blood-brain barrier (BBB) protects the brain against potentially neurotoxic molecules in the circulation, and loss of its integrity may contribute to disease progression in neurodegenerative conditions. Recently, the active role of reactive astrocytes in BBB disruption has become evident in the inflamed brain. In the present study, we investigated whether mesenchymal stem cell (MSC) treatment might modulate reactive astrocytes and thus stabilize BBB integrity through vascular endothelial growth factor A (VEGF-A) signaling in inflammatory conditions.

Methods: For the inflamed brain, we injected LPS using a stereotaxic apparatus and MSCs were injected into the tail vein. At 6 hours and 7 days after LPS injection, we analyzed modulatory effects of MSCs on the change of BBB permeability through VEGF-A signaling using immunochemistry and western blot. To determine the effects of MSCs on VEGF-A-related signaling in cellular system, we had used endothelial cells treated with VEGF-A and co-cultured astrocyte and BV 2 cells treated with lipopolysaccharide (LPS) and then these cells were co-cultured with MSCs.

Results: In LPS-treated rats, MSCs restored Evans blue infiltration and the number of endothelial-barrier antigen (EBA) and P-glycoprotein (p-gp)-expressing cells, which were significantly altered in LPS-treated animals. Additionally, MSC administration following LPS treatment markedly increased the density of astrocytic filaments around vessels and reversed LPS-induced elevations in VEGF-A levels as well as endothelial nitric oxide synthase (eNOS)-dependent downregulation of tight junction proteins in the endothelium. Consequently, MSC treatment reduced neutrophil infiltration and enhanced survival of midbrain dopaminergic neurons in LPS-treated animals. In cellular system, MSC treatment led to a significant reversion of VEGF-A-induced eNOS and tight junction protein expression in endothelial cells, which led to increased EBA expressing cells. Additionally, MSC treatment significantly attenuated LPS-induced increased expressions of IL-1β in microglia and VEGF-A in astrocytes with an increase in IL-10 levels.

Conclusion: The present study indicated that MSCs may stabilize BBB permeability by modulating astrocytic endfeet and VEGF-A signaling, which may be relevant to the treatment of Parkinsonian diseases as a candidate for disease modifying therapeutics.

No MeSH data available.


Related in: MedlinePlus