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S1P, dihydro-S1P and C24:1-ceramide levels in the HDL-containing fraction of serum inversely correlate with occurrence of ischemic heart disease.

Argraves KM, Sethi AA, Gazzolo PJ, Wilkerson BA, Remaley AT, Tybjaerg-Hansen A, Nordestgaard BG, Yeatts SD, Nicholas KS, Barth JL, Argraves WS - Lipids Health Dis (2011)

Bottom Line: Emerging evidence suggests that many of the effects of HDL on cardiovascular function may be attributable to its S1P cargo.The results show a highly significant inverse relationship between the level of S1P in the HDL-containing fraction of serum and the occurrence of IHD.These findings indicate that compositional differences of sphingolipids in the HDL-containing fraction of human serum are related to the occurrence of IHD, and may contribute to the putative protective role of HDL in IHD.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC 29425, USA. argravek@musc.edu

ABSTRACT

Background: The lysosphingolipid sphingosine 1-phosphate (S1P) is carried in the blood in association with lipoproteins, predominantly high density lipoproteins (HDL). Emerging evidence suggests that many of the effects of HDL on cardiovascular function may be attributable to its S1P cargo.

Methods: Here we have evaluated how levels of S1P and related sphingolipids in an HDL-containing fraction of human serum correlate with occurrence of ischemic heart disease (IHD). To accomplish this we used liquid chromatography-mass spectrometry to measure S1P levels in the HDL-containing fraction of serum (depleted of LDL and VLDL) from 204 subjects in the Copenhagen City Heart Study (CCHS). The study group consisted of individuals having high serum HDL cholesterol (HDL-C) (females:≥ 73.5 mg/dL; males:≥ 61.9 mg/dL) and verified IHD; subjects with high HDL-C and no IHD; individuals with low HDL-C (females:≤ 38.7 mg/dL; males:≤ 34.1 mg/dL) and IHD, and subjects with low HDL-C and no IHD.

Results: The results show a highly significant inverse relationship between the level of S1P in the HDL-containing fraction of serum and the occurrence of IHD. Furthermore, an inverse relationship with IHD was also observed for two other sphingolipids, dihydro-S1P and C24:1-ceramide, in the HDL-containing fraction of serum. Additionally, we demonstrated that the amount of S1P on HDL correlates with the magnitude of HDL-induced endothelial cell barrier signaling.

Conclusions: These findings indicate that compositional differences of sphingolipids in the HDL-containing fraction of human serum are related to the occurrence of IHD, and may contribute to the putative protective role of HDL in IHD.

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DH-S1P enhances endothelial barrier in an S1P1 dependent manner. Confluent EC monolayers were grown under serum-free conditions until a minimal TEER plateau was reached. In A, EC monolayers were incubated with varying concentrations of DH-S1P [111-1000 nM]. In B, EC monolayers were incubated with varying concentrations of S1P [111-1000 nM]. In C, EC monolayers were incubated with DH-S1P [1000 nM] plus and minus the S1P1 antagonist W146 or the S1P1/S1P3 antagonist VPC23019 (each at 10 μM). In D, EC monolayers were incubated with S1P [1000 nM] plus and minus the S1P1 antagonist W146 or the S1P1/S1P3 antagonist VPC23019 (each at 10 μM). Each of the TEER tracings shown is an average from three independent experiments each with two replicates per condition. Impedance values were normalized by dividing each value by the level of impedance measured just prior to the addition of effectors. As a control for A and B, monolayers were treated with 40 μg/ml delipidated albumin (Control), a concentration corresponding to the amount of BSA carrier used for the highest concentration of DH-S1P and S1P tested. As controls for C and D, ECs were treated with delipidated albumin-containing serum free medium (SFM) plus vehicle buffer.
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Figure 3: DH-S1P enhances endothelial barrier in an S1P1 dependent manner. Confluent EC monolayers were grown under serum-free conditions until a minimal TEER plateau was reached. In A, EC monolayers were incubated with varying concentrations of DH-S1P [111-1000 nM]. In B, EC monolayers were incubated with varying concentrations of S1P [111-1000 nM]. In C, EC monolayers were incubated with DH-S1P [1000 nM] plus and minus the S1P1 antagonist W146 or the S1P1/S1P3 antagonist VPC23019 (each at 10 μM). In D, EC monolayers were incubated with S1P [1000 nM] plus and minus the S1P1 antagonist W146 or the S1P1/S1P3 antagonist VPC23019 (each at 10 μM). Each of the TEER tracings shown is an average from three independent experiments each with two replicates per condition. Impedance values were normalized by dividing each value by the level of impedance measured just prior to the addition of effectors. As a control for A and B, monolayers were treated with 40 μg/ml delipidated albumin (Control), a concentration corresponding to the amount of BSA carrier used for the highest concentration of DH-S1P and S1P tested. As controls for C and D, ECs were treated with delipidated albumin-containing serum free medium (SFM) plus vehicle buffer.

Mentions: While the effects of S1P on endothelial cell functions have been implicated as a basis for the anti-atherogenic effects of HDL, rather little is known as to the effects of DH-S1P on vascular cell behaviors. Generally, DH-S1P has been found to be an agonist of S1P receptors [11-13]. We used electrical cell substrate impedance sensing (ECIS) to assess the ability of DH-S1P to influence endothelial barrier activity, a major physiological function of the endothelium. DH-S1P (on an albumin carrier) induced a dose-dependent increase in transendothelial electrical resistance (TEER) (Figure 3A). The magnitude of the response was similar to that achieved using S1P (Figure 3B). By contrast, C24:1-ceramide, the third sphingolipid found to be significantly lower in HDL-containing fractions of serum from individuals with IHD, did not effect TEER (Additional file 1, Figure S1).


S1P, dihydro-S1P and C24:1-ceramide levels in the HDL-containing fraction of serum inversely correlate with occurrence of ischemic heart disease.

Argraves KM, Sethi AA, Gazzolo PJ, Wilkerson BA, Remaley AT, Tybjaerg-Hansen A, Nordestgaard BG, Yeatts SD, Nicholas KS, Barth JL, Argraves WS - Lipids Health Dis (2011)

DH-S1P enhances endothelial barrier in an S1P1 dependent manner. Confluent EC monolayers were grown under serum-free conditions until a minimal TEER plateau was reached. In A, EC monolayers were incubated with varying concentrations of DH-S1P [111-1000 nM]. In B, EC monolayers were incubated with varying concentrations of S1P [111-1000 nM]. In C, EC monolayers were incubated with DH-S1P [1000 nM] plus and minus the S1P1 antagonist W146 or the S1P1/S1P3 antagonist VPC23019 (each at 10 μM). In D, EC monolayers were incubated with S1P [1000 nM] plus and minus the S1P1 antagonist W146 or the S1P1/S1P3 antagonist VPC23019 (each at 10 μM). Each of the TEER tracings shown is an average from three independent experiments each with two replicates per condition. Impedance values were normalized by dividing each value by the level of impedance measured just prior to the addition of effectors. As a control for A and B, monolayers were treated with 40 μg/ml delipidated albumin (Control), a concentration corresponding to the amount of BSA carrier used for the highest concentration of DH-S1P and S1P tested. As controls for C and D, ECs were treated with delipidated albumin-containing serum free medium (SFM) plus vehicle buffer.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 3: DH-S1P enhances endothelial barrier in an S1P1 dependent manner. Confluent EC monolayers were grown under serum-free conditions until a minimal TEER plateau was reached. In A, EC monolayers were incubated with varying concentrations of DH-S1P [111-1000 nM]. In B, EC monolayers were incubated with varying concentrations of S1P [111-1000 nM]. In C, EC monolayers were incubated with DH-S1P [1000 nM] plus and minus the S1P1 antagonist W146 or the S1P1/S1P3 antagonist VPC23019 (each at 10 μM). In D, EC monolayers were incubated with S1P [1000 nM] plus and minus the S1P1 antagonist W146 or the S1P1/S1P3 antagonist VPC23019 (each at 10 μM). Each of the TEER tracings shown is an average from three independent experiments each with two replicates per condition. Impedance values were normalized by dividing each value by the level of impedance measured just prior to the addition of effectors. As a control for A and B, monolayers were treated with 40 μg/ml delipidated albumin (Control), a concentration corresponding to the amount of BSA carrier used for the highest concentration of DH-S1P and S1P tested. As controls for C and D, ECs were treated with delipidated albumin-containing serum free medium (SFM) plus vehicle buffer.
Mentions: While the effects of S1P on endothelial cell functions have been implicated as a basis for the anti-atherogenic effects of HDL, rather little is known as to the effects of DH-S1P on vascular cell behaviors. Generally, DH-S1P has been found to be an agonist of S1P receptors [11-13]. We used electrical cell substrate impedance sensing (ECIS) to assess the ability of DH-S1P to influence endothelial barrier activity, a major physiological function of the endothelium. DH-S1P (on an albumin carrier) induced a dose-dependent increase in transendothelial electrical resistance (TEER) (Figure 3A). The magnitude of the response was similar to that achieved using S1P (Figure 3B). By contrast, C24:1-ceramide, the third sphingolipid found to be significantly lower in HDL-containing fractions of serum from individuals with IHD, did not effect TEER (Additional file 1, Figure S1).

Bottom Line: Emerging evidence suggests that many of the effects of HDL on cardiovascular function may be attributable to its S1P cargo.The results show a highly significant inverse relationship between the level of S1P in the HDL-containing fraction of serum and the occurrence of IHD.These findings indicate that compositional differences of sphingolipids in the HDL-containing fraction of human serum are related to the occurrence of IHD, and may contribute to the putative protective role of HDL in IHD.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC 29425, USA. argravek@musc.edu

ABSTRACT

Background: The lysosphingolipid sphingosine 1-phosphate (S1P) is carried in the blood in association with lipoproteins, predominantly high density lipoproteins (HDL). Emerging evidence suggests that many of the effects of HDL on cardiovascular function may be attributable to its S1P cargo.

Methods: Here we have evaluated how levels of S1P and related sphingolipids in an HDL-containing fraction of human serum correlate with occurrence of ischemic heart disease (IHD). To accomplish this we used liquid chromatography-mass spectrometry to measure S1P levels in the HDL-containing fraction of serum (depleted of LDL and VLDL) from 204 subjects in the Copenhagen City Heart Study (CCHS). The study group consisted of individuals having high serum HDL cholesterol (HDL-C) (females:≥ 73.5 mg/dL; males:≥ 61.9 mg/dL) and verified IHD; subjects with high HDL-C and no IHD; individuals with low HDL-C (females:≤ 38.7 mg/dL; males:≤ 34.1 mg/dL) and IHD, and subjects with low HDL-C and no IHD.

Results: The results show a highly significant inverse relationship between the level of S1P in the HDL-containing fraction of serum and the occurrence of IHD. Furthermore, an inverse relationship with IHD was also observed for two other sphingolipids, dihydro-S1P and C24:1-ceramide, in the HDL-containing fraction of serum. Additionally, we demonstrated that the amount of S1P on HDL correlates with the magnitude of HDL-induced endothelial cell barrier signaling.

Conclusions: These findings indicate that compositional differences of sphingolipids in the HDL-containing fraction of human serum are related to the occurrence of IHD, and may contribute to the putative protective role of HDL in IHD.

Show MeSH
Related in: MedlinePlus