Limits...
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

MSCs modulate tight junction protein expression via VEGF-A in LPS-induced animals. Compared with controls, the VEGF-A levels were significantly increased from 12 hours after LPS injection, and elevated VEGF-A levels were sustained 7 days after LPS injection. Meanwhile, MSC treatment significantly attenuated VEGF-A expressions in LPS-treated rats a (n = 3). *p <0.05, **p <0.01. Double immunofluorescence showed that LPS treatment markedly upregulated eNOS immunoreactivity b and downregulated CLN-5 immunoreactivity c in the endothelial cells of the SN, whereas MSC treatment attenuated eNOS expression and increased CLN-5 expression in the endothelial cells at 12 hours and 7 days after LPS injection b, c. Scale bar: 20 μm. CLN-5 claudin-5, DAPI 4',6-diamidino-2-phenylindole, eNOS endothelial nitric oxide synthase, LPS lipopolysaccharide, MSC mesenchymal stem cell, VEGF vascular endothelial growth factor
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4588687&req=5

Fig3: MSCs modulate tight junction protein expression via VEGF-A in LPS-induced animals. Compared with controls, the VEGF-A levels were significantly increased from 12 hours after LPS injection, and elevated VEGF-A levels were sustained 7 days after LPS injection. Meanwhile, MSC treatment significantly attenuated VEGF-A expressions in LPS-treated rats a (n = 3). *p <0.05, **p <0.01. Double immunofluorescence showed that LPS treatment markedly upregulated eNOS immunoreactivity b and downregulated CLN-5 immunoreactivity c in the endothelial cells of the SN, whereas MSC treatment attenuated eNOS expression and increased CLN-5 expression in the endothelial cells at 12 hours and 7 days after LPS injection b, c. Scale bar: 20 μm. CLN-5 claudin-5, DAPI 4',6-diamidino-2-phenylindole, eNOS endothelial nitric oxide synthase, LPS lipopolysaccharide, MSC mesenchymal stem cell, VEGF vascular endothelial growth factor

Mentions: To evaluate the effects of MSCs on expression of VEGF-A, we determined VEGF-A levels by ELISA at 6 hours, 12 hours, 1 day, 2 days, and 7 days after LPS injection. Compared with controls, LPS-treated rats expressed significantly greater levels of VEGF-A from 12 hours after LPS injection, and elevated VEGF-A levels were sustained 7 days after LPS injection. However, MSC treatment significantly reduced VEGF-A expression compared with that in LPS only-treated rats, and these levels were similar to those in controls (Fig. 3a). Furthermore, we evaluated whether the effects of MSCs on VEGF-A regulate eNOS expression and tight junction integrity by double immunostaining for eNOS and CD31 or CLN-5 and CD31. LPS treatment markedly upregulated eNOS immunoreactivity and downregulated CLN-5 immunoreactivity in the endothelial cells of the SN; however, MSC treatment in LPS-treated animals attenuated eNOS expression and increased CLN-5 expression in the endothelial cells at 12 hours and 7 days after LPS injection (Fig. 3b, c).Fig. 3


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)

MSCs modulate tight junction protein expression via VEGF-A in LPS-induced animals. Compared with controls, the VEGF-A levels were significantly increased from 12 hours after LPS injection, and elevated VEGF-A levels were sustained 7 days after LPS injection. Meanwhile, MSC treatment significantly attenuated VEGF-A expressions in LPS-treated rats a (n = 3). *p <0.05, **p <0.01. Double immunofluorescence showed that LPS treatment markedly upregulated eNOS immunoreactivity b and downregulated CLN-5 immunoreactivity c in the endothelial cells of the SN, whereas MSC treatment attenuated eNOS expression and increased CLN-5 expression in the endothelial cells at 12 hours and 7 days after LPS injection b, c. Scale bar: 20 μm. CLN-5 claudin-5, DAPI 4',6-diamidino-2-phenylindole, eNOS endothelial nitric oxide synthase, LPS lipopolysaccharide, MSC mesenchymal stem cell, VEGF vascular endothelial growth factor
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4588687&req=5

Fig3: MSCs modulate tight junction protein expression via VEGF-A in LPS-induced animals. Compared with controls, the VEGF-A levels were significantly increased from 12 hours after LPS injection, and elevated VEGF-A levels were sustained 7 days after LPS injection. Meanwhile, MSC treatment significantly attenuated VEGF-A expressions in LPS-treated rats a (n = 3). *p <0.05, **p <0.01. Double immunofluorescence showed that LPS treatment markedly upregulated eNOS immunoreactivity b and downregulated CLN-5 immunoreactivity c in the endothelial cells of the SN, whereas MSC treatment attenuated eNOS expression and increased CLN-5 expression in the endothelial cells at 12 hours and 7 days after LPS injection b, c. Scale bar: 20 μm. CLN-5 claudin-5, DAPI 4',6-diamidino-2-phenylindole, eNOS endothelial nitric oxide synthase, LPS lipopolysaccharide, MSC mesenchymal stem cell, VEGF vascular endothelial growth factor
Mentions: To evaluate the effects of MSCs on expression of VEGF-A, we determined VEGF-A levels by ELISA at 6 hours, 12 hours, 1 day, 2 days, and 7 days after LPS injection. Compared with controls, LPS-treated rats expressed significantly greater levels of VEGF-A from 12 hours after LPS injection, and elevated VEGF-A levels were sustained 7 days after LPS injection. However, MSC treatment significantly reduced VEGF-A expression compared with that in LPS only-treated rats, and these levels were similar to those in controls (Fig. 3a). Furthermore, we evaluated whether the effects of MSCs on VEGF-A regulate eNOS expression and tight junction integrity by double immunostaining for eNOS and CD31 or CLN-5 and CD31. LPS treatment markedly upregulated eNOS immunoreactivity and downregulated CLN-5 immunoreactivity in the endothelial cells of the SN; however, MSC treatment in LPS-treated animals attenuated eNOS expression and increased CLN-5 expression in the endothelial cells at 12 hours and 7 days after LPS injection (Fig. 3b, c).Fig. 3

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