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Critical role of sphingosine-1-phosphate receptor-2 in the disruption of cerebrovascular integrity in experimental stroke.

Kim GS, Yang L, Zhang G, Zhao H, Selim M, McCullough LD, Kluk MJ, Sanchez T - Nat Commun (2015)

Bottom Line: In addition, inhibition of S1PR2 results in decreased matrix metalloproteinase (MMP)-9 activity in vivo and lower gelatinase activity in cerebral microvessels.S1PR2 immunopositivity is detected only in the ischemic microvessels of wild-type mice and in the cerebrovascular endothelium of human brain autopsy samples.In vitro, S1PR2 potently regulates the responses of the brain endothelium to ischaemic and inflammatory injury.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Emergency Medicine, the Center for Vascular Biology Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA [2] Department of Surgery, the Center for Vascular Biology Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.

ABSTRACT
The use and effectiveness of current stroke reperfusion therapies are limited by the complications of reperfusion injury, which include increased cerebrovascular permeability and haemorrhagic transformation. Sphingosine-1-phosphate (S1P) is emerging as a potent modulator of vascular integrity via its receptors (S1PR). By using genetic approaches and a S1PR2 antagonist (JTE013), here we show that S1PR2 plays a critical role in the induction of cerebrovascular permeability, development of intracerebral haemorrhage and neurovascular injury in experimental stroke. In addition, inhibition of S1PR2 results in decreased matrix metalloproteinase (MMP)-9 activity in vivo and lower gelatinase activity in cerebral microvessels. S1PR2 immunopositivity is detected only in the ischemic microvessels of wild-type mice and in the cerebrovascular endothelium of human brain autopsy samples. In vitro, S1PR2 potently regulates the responses of the brain endothelium to ischaemic and inflammatory injury. Therapeutic targeting of this novel pathway could have important translational relevance to stroke patients.

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Critical role of S1PR2 in the disruption of cerebrovascular integrity after I/R injuryA, B) Cerebrovascular permeability (vasogenic edema) was determined by Evans Blue Dye (EBD) extravasation assay, 3 hours after reperfusion in S1pr2+/+, S1pr2−/− and S1pr2+/+ receiving JTE013. A) Brain coronal slices of representative animals. Scale bar 5 mm. B) Quantification of extravascular EBD. Ipsilateral/contralateral (IL/CL) ratios are shown. C,D) Hemorrhagic transformation was assessed 24 hours after reperfusion in S1pr2+/+, S1pr2−/− and S1pr2+/+ receiving JTE013. C) Representative images are shown. Scale bar 5 mm. D) Quantification of extravascular hemoglobin (Hb) content. Values are ipsilateral/contralateral (IL/CL) ratios of the μg of Hb. B, D) The individual values and the mean ± SEM are shown, n=5–8 from 4 independent experiments. *p<0.05 (one-way ANOVA followed by Newman-Keuls).
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Figure 3: Critical role of S1PR2 in the disruption of cerebrovascular integrity after I/R injuryA, B) Cerebrovascular permeability (vasogenic edema) was determined by Evans Blue Dye (EBD) extravasation assay, 3 hours after reperfusion in S1pr2+/+, S1pr2−/− and S1pr2+/+ receiving JTE013. A) Brain coronal slices of representative animals. Scale bar 5 mm. B) Quantification of extravascular EBD. Ipsilateral/contralateral (IL/CL) ratios are shown. C,D) Hemorrhagic transformation was assessed 24 hours after reperfusion in S1pr2+/+, S1pr2−/− and S1pr2+/+ receiving JTE013. C) Representative images are shown. Scale bar 5 mm. D) Quantification of extravascular hemoglobin (Hb) content. Values are ipsilateral/contralateral (IL/CL) ratios of the μg of Hb. B, D) The individual values and the mean ± SEM are shown, n=5–8 from 4 independent experiments. *p<0.05 (one-way ANOVA followed by Newman-Keuls).

Mentions: During cerebral ischemia, blood brain barrier disruption starts early after the onset of ischemia and increased cerebrovascular permeability can be detected as early as 3 hours after reperfusion 25,26. In order to investigate the role of S1PR2 in vasogenic edema (increased cerebrovascular permeability), we conducted Evans Blue Dye (EBD) extravasation assays. As shown in Figure 3A and B, S1pr2−/− mice exhibited a dramatic decrease in EBD extravasation, 3 hours after reperfusion, compared to their wild type litter mates (ratio of EBD content in ipsilateral hemisphere/contralateral hemisphere was 1.6±0.31 in S1pr2−/− mice vs 4.54±0.83 in wild type). In addition, acute inhibition of S1PR2 signaling in wild type mice, by JTE013 administration, potently blocked the induction of cerebrovascular permeability after I/R injury (ratio of EBD content 1.28±0.08, Figure 3A, B).


Critical role of sphingosine-1-phosphate receptor-2 in the disruption of cerebrovascular integrity in experimental stroke.

Kim GS, Yang L, Zhang G, Zhao H, Selim M, McCullough LD, Kluk MJ, Sanchez T - Nat Commun (2015)

Critical role of S1PR2 in the disruption of cerebrovascular integrity after I/R injuryA, B) Cerebrovascular permeability (vasogenic edema) was determined by Evans Blue Dye (EBD) extravasation assay, 3 hours after reperfusion in S1pr2+/+, S1pr2−/− and S1pr2+/+ receiving JTE013. A) Brain coronal slices of representative animals. Scale bar 5 mm. B) Quantification of extravascular EBD. Ipsilateral/contralateral (IL/CL) ratios are shown. C,D) Hemorrhagic transformation was assessed 24 hours after reperfusion in S1pr2+/+, S1pr2−/− and S1pr2+/+ receiving JTE013. C) Representative images are shown. Scale bar 5 mm. D) Quantification of extravascular hemoglobin (Hb) content. Values are ipsilateral/contralateral (IL/CL) ratios of the μg of Hb. B, D) The individual values and the mean ± SEM are shown, n=5–8 from 4 independent experiments. *p<0.05 (one-way ANOVA followed by Newman-Keuls).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4587559&req=5

Figure 3: Critical role of S1PR2 in the disruption of cerebrovascular integrity after I/R injuryA, B) Cerebrovascular permeability (vasogenic edema) was determined by Evans Blue Dye (EBD) extravasation assay, 3 hours after reperfusion in S1pr2+/+, S1pr2−/− and S1pr2+/+ receiving JTE013. A) Brain coronal slices of representative animals. Scale bar 5 mm. B) Quantification of extravascular EBD. Ipsilateral/contralateral (IL/CL) ratios are shown. C,D) Hemorrhagic transformation was assessed 24 hours after reperfusion in S1pr2+/+, S1pr2−/− and S1pr2+/+ receiving JTE013. C) Representative images are shown. Scale bar 5 mm. D) Quantification of extravascular hemoglobin (Hb) content. Values are ipsilateral/contralateral (IL/CL) ratios of the μg of Hb. B, D) The individual values and the mean ± SEM are shown, n=5–8 from 4 independent experiments. *p<0.05 (one-way ANOVA followed by Newman-Keuls).
Mentions: During cerebral ischemia, blood brain barrier disruption starts early after the onset of ischemia and increased cerebrovascular permeability can be detected as early as 3 hours after reperfusion 25,26. In order to investigate the role of S1PR2 in vasogenic edema (increased cerebrovascular permeability), we conducted Evans Blue Dye (EBD) extravasation assays. As shown in Figure 3A and B, S1pr2−/− mice exhibited a dramatic decrease in EBD extravasation, 3 hours after reperfusion, compared to their wild type litter mates (ratio of EBD content in ipsilateral hemisphere/contralateral hemisphere was 1.6±0.31 in S1pr2−/− mice vs 4.54±0.83 in wild type). In addition, acute inhibition of S1PR2 signaling in wild type mice, by JTE013 administration, potently blocked the induction of cerebrovascular permeability after I/R injury (ratio of EBD content 1.28±0.08, Figure 3A, B).

Bottom Line: In addition, inhibition of S1PR2 results in decreased matrix metalloproteinase (MMP)-9 activity in vivo and lower gelatinase activity in cerebral microvessels.S1PR2 immunopositivity is detected only in the ischemic microvessels of wild-type mice and in the cerebrovascular endothelium of human brain autopsy samples.In vitro, S1PR2 potently regulates the responses of the brain endothelium to ischaemic and inflammatory injury.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Emergency Medicine, the Center for Vascular Biology Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA [2] Department of Surgery, the Center for Vascular Biology Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.

ABSTRACT
The use and effectiveness of current stroke reperfusion therapies are limited by the complications of reperfusion injury, which include increased cerebrovascular permeability and haemorrhagic transformation. Sphingosine-1-phosphate (S1P) is emerging as a potent modulator of vascular integrity via its receptors (S1PR). By using genetic approaches and a S1PR2 antagonist (JTE013), here we show that S1PR2 plays a critical role in the induction of cerebrovascular permeability, development of intracerebral haemorrhage and neurovascular injury in experimental stroke. In addition, inhibition of S1PR2 results in decreased matrix metalloproteinase (MMP)-9 activity in vivo and lower gelatinase activity in cerebral microvessels. S1PR2 immunopositivity is detected only in the ischemic microvessels of wild-type mice and in the cerebrovascular endothelium of human brain autopsy samples. In vitro, S1PR2 potently regulates the responses of the brain endothelium to ischaemic and inflammatory injury. Therapeutic targeting of this novel pathway could have important translational relevance to stroke patients.

Show MeSH
Related in: MedlinePlus