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The biphasic effects of oxidized-low density lipoprotein on the vasculogenic function of endothelial progenitor cells.

Lin FY, Tsao NW, Shih CM, Lin YW, Yeh JS, Chen JW, Nakagami H, Morishita R, Sawamura T, Huang CY - PLoS ONE (2015)

Bottom Line: This study aimed to investigate the concentration-related effects of oxLDL on EPC functions and related angiogenesis, in vitro and in vivo.Higher concentrations of oxLDL (10-50 μg/mL) impaired EPC function via the activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase pathways and consequent inhibition of eNOS activity, which could be reversed by anti-oxidants (diphenylene iodonium and apocynin) and gp91phox siRNA.In conclusion, oxLDL has concentration-dependent biphasic effects on human late-outgrowth EPC tube formation in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan; Division of Cardiology, Department of Internal Medicine and Cardiovascular Research Center, Taipei Medical University Hospital, Taipei, Taiwan.

ABSTRACT
Late-outgrowth endothelial progenitor cells (EPCs) are stress-resistant and responsible for reparative functions in the cardiovascular system. Oxidized-LDL (oxLDL) plays a critical role in cardiovascular disease pathogenesis. However, it is largely unknown what the impacts of oxLDL are on late-outgrowth EPCs. This study aimed to investigate the concentration-related effects of oxLDL on EPC functions and related angiogenesis, in vitro and in vivo. In this study, early and late-outgrowth EPCs were generated from circulating human mononuclear cells. oxLDL may regulate EPC vasculogenic function via the lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1). Lower concentrations (5 μg/mL) of oxLDL can potentiate EPC tube formation in vitro and in vivo by activating eNOS mechanisms, which are mediated by p38 MAPK- and SAPK/JNK-related pathways. Higher concentrations of oxLDL (10-50 μg/mL) impaired EPC function via the activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase pathways and consequent inhibition of eNOS activity, which could be reversed by anti-oxidants (diphenylene iodonium and apocynin) and gp91phox siRNA. In conclusion, oxLDL has concentration-dependent biphasic effects on human late-outgrowth EPC tube formation in vitro and in vivo.

No MeSH data available.


Related in: MedlinePlus

p38 MAPK- and SAPK/JNK-related pathways contribute to increased tube formation in EPCs under low concentrated oxLDL.(A) Following treatment of EPCs with 0–50 μg/mL of oxLDL for 4 hours, the phosphorylation of p38 MAPK, SAPK/JNK and ERK1/2 were analyzed by Western blot. The total p38 MAPK, SAPK/JNK, ERK1/2, and β-actin protein levels were used as loading controls. The graph showed the quantitative activation of p38 MAPK (phospho-p38MAPK/total- p38MAPK ratio), SAPK/JNK (phospho-SAPK/JNK/total-SAPK/JNK ratio), and Akt (phospho-ERK1/2/total-ERK1/2 ratio) density in oxLDL-treated EPCs. (B) EPCs were pretreated with SP600125, PD98059, or SB203580 for 1 hour prior to treatment with 5 or 50 μg/mL oxLDL for 24 hours. (C) Treatment of SP600125, PD98059, or SB203580 alone for 24 hours. In vitro angiogenesis was assayed using ECMatrix gel. (D) EPCs were pretreated with SP600125 or SB203580 for 1 hour prior to treatment with 5 μg/mL oxLDL for 24 hours. The NO-containing conditional medium was analyzed using electron spin resonance spectroscopy. Data are expressed as the mean ± SEM of three experiments performed in triplicate. *p < 0.05 was considered significant. (E) EPCs were pretreated with SP600125 or SB203580 for 1 hour prior to treatment with 5 μg/mL oxLDL for 2 hours, subsequently Akt activation (phosphorylation) was analyzed by Western blot. Total Akt protein levels were used as loading controls.
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pone.0123971.g004: p38 MAPK- and SAPK/JNK-related pathways contribute to increased tube formation in EPCs under low concentrated oxLDL.(A) Following treatment of EPCs with 0–50 μg/mL of oxLDL for 4 hours, the phosphorylation of p38 MAPK, SAPK/JNK and ERK1/2 were analyzed by Western blot. The total p38 MAPK, SAPK/JNK, ERK1/2, and β-actin protein levels were used as loading controls. The graph showed the quantitative activation of p38 MAPK (phospho-p38MAPK/total- p38MAPK ratio), SAPK/JNK (phospho-SAPK/JNK/total-SAPK/JNK ratio), and Akt (phospho-ERK1/2/total-ERK1/2 ratio) density in oxLDL-treated EPCs. (B) EPCs were pretreated with SP600125, PD98059, or SB203580 for 1 hour prior to treatment with 5 or 50 μg/mL oxLDL for 24 hours. (C) Treatment of SP600125, PD98059, or SB203580 alone for 24 hours. In vitro angiogenesis was assayed using ECMatrix gel. (D) EPCs were pretreated with SP600125 or SB203580 for 1 hour prior to treatment with 5 μg/mL oxLDL for 24 hours. The NO-containing conditional medium was analyzed using electron spin resonance spectroscopy. Data are expressed as the mean ± SEM of three experiments performed in triplicate. *p < 0.05 was considered significant. (E) EPCs were pretreated with SP600125 or SB203580 for 1 hour prior to treatment with 5 μg/mL oxLDL for 2 hours, subsequently Akt activation (phosphorylation) was analyzed by Western blot. Total Akt protein levels were used as loading controls.

Mentions: The potential roles of mitogen-activated protein kinase (MAPK)-related mechanisms were also examined. OxLDL markedly induced the phosphorylation of MAPKs, including p38 MAPK and SAPK/JNK (Fig 4A), after exposure to 0.1–5 μg/mL of oxLDL for 4 hours. Interestingly, exposure to 10–50 μg/mL of oxLDL did not change the phosphorylation of p38 MAPK, ERK1/2, and SAPK/JNK in EPCs.


The biphasic effects of oxidized-low density lipoprotein on the vasculogenic function of endothelial progenitor cells.

Lin FY, Tsao NW, Shih CM, Lin YW, Yeh JS, Chen JW, Nakagami H, Morishita R, Sawamura T, Huang CY - PLoS ONE (2015)

p38 MAPK- and SAPK/JNK-related pathways contribute to increased tube formation in EPCs under low concentrated oxLDL.(A) Following treatment of EPCs with 0–50 μg/mL of oxLDL for 4 hours, the phosphorylation of p38 MAPK, SAPK/JNK and ERK1/2 were analyzed by Western blot. The total p38 MAPK, SAPK/JNK, ERK1/2, and β-actin protein levels were used as loading controls. The graph showed the quantitative activation of p38 MAPK (phospho-p38MAPK/total- p38MAPK ratio), SAPK/JNK (phospho-SAPK/JNK/total-SAPK/JNK ratio), and Akt (phospho-ERK1/2/total-ERK1/2 ratio) density in oxLDL-treated EPCs. (B) EPCs were pretreated with SP600125, PD98059, or SB203580 for 1 hour prior to treatment with 5 or 50 μg/mL oxLDL for 24 hours. (C) Treatment of SP600125, PD98059, or SB203580 alone for 24 hours. In vitro angiogenesis was assayed using ECMatrix gel. (D) EPCs were pretreated with SP600125 or SB203580 for 1 hour prior to treatment with 5 μg/mL oxLDL for 24 hours. The NO-containing conditional medium was analyzed using electron spin resonance spectroscopy. Data are expressed as the mean ± SEM of three experiments performed in triplicate. *p < 0.05 was considered significant. (E) EPCs were pretreated with SP600125 or SB203580 for 1 hour prior to treatment with 5 μg/mL oxLDL for 2 hours, subsequently Akt activation (phosphorylation) was analyzed by Western blot. Total Akt protein levels were used as loading controls.
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pone.0123971.g004: p38 MAPK- and SAPK/JNK-related pathways contribute to increased tube formation in EPCs under low concentrated oxLDL.(A) Following treatment of EPCs with 0–50 μg/mL of oxLDL for 4 hours, the phosphorylation of p38 MAPK, SAPK/JNK and ERK1/2 were analyzed by Western blot. The total p38 MAPK, SAPK/JNK, ERK1/2, and β-actin protein levels were used as loading controls. The graph showed the quantitative activation of p38 MAPK (phospho-p38MAPK/total- p38MAPK ratio), SAPK/JNK (phospho-SAPK/JNK/total-SAPK/JNK ratio), and Akt (phospho-ERK1/2/total-ERK1/2 ratio) density in oxLDL-treated EPCs. (B) EPCs were pretreated with SP600125, PD98059, or SB203580 for 1 hour prior to treatment with 5 or 50 μg/mL oxLDL for 24 hours. (C) Treatment of SP600125, PD98059, or SB203580 alone for 24 hours. In vitro angiogenesis was assayed using ECMatrix gel. (D) EPCs were pretreated with SP600125 or SB203580 for 1 hour prior to treatment with 5 μg/mL oxLDL for 24 hours. The NO-containing conditional medium was analyzed using electron spin resonance spectroscopy. Data are expressed as the mean ± SEM of three experiments performed in triplicate. *p < 0.05 was considered significant. (E) EPCs were pretreated with SP600125 or SB203580 for 1 hour prior to treatment with 5 μg/mL oxLDL for 2 hours, subsequently Akt activation (phosphorylation) was analyzed by Western blot. Total Akt protein levels were used as loading controls.
Mentions: The potential roles of mitogen-activated protein kinase (MAPK)-related mechanisms were also examined. OxLDL markedly induced the phosphorylation of MAPKs, including p38 MAPK and SAPK/JNK (Fig 4A), after exposure to 0.1–5 μg/mL of oxLDL for 4 hours. Interestingly, exposure to 10–50 μg/mL of oxLDL did not change the phosphorylation of p38 MAPK, ERK1/2, and SAPK/JNK in EPCs.

Bottom Line: This study aimed to investigate the concentration-related effects of oxLDL on EPC functions and related angiogenesis, in vitro and in vivo.Higher concentrations of oxLDL (10-50 μg/mL) impaired EPC function via the activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase pathways and consequent inhibition of eNOS activity, which could be reversed by anti-oxidants (diphenylene iodonium and apocynin) and gp91phox siRNA.In conclusion, oxLDL has concentration-dependent biphasic effects on human late-outgrowth EPC tube formation in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan; Division of Cardiology, Department of Internal Medicine and Cardiovascular Research Center, Taipei Medical University Hospital, Taipei, Taiwan.

ABSTRACT
Late-outgrowth endothelial progenitor cells (EPCs) are stress-resistant and responsible for reparative functions in the cardiovascular system. Oxidized-LDL (oxLDL) plays a critical role in cardiovascular disease pathogenesis. However, it is largely unknown what the impacts of oxLDL are on late-outgrowth EPCs. This study aimed to investigate the concentration-related effects of oxLDL on EPC functions and related angiogenesis, in vitro and in vivo. In this study, early and late-outgrowth EPCs were generated from circulating human mononuclear cells. oxLDL may regulate EPC vasculogenic function via the lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1). Lower concentrations (5 μg/mL) of oxLDL can potentiate EPC tube formation in vitro and in vivo by activating eNOS mechanisms, which are mediated by p38 MAPK- and SAPK/JNK-related pathways. Higher concentrations of oxLDL (10-50 μg/mL) impaired EPC function via the activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase pathways and consequent inhibition of eNOS activity, which could be reversed by anti-oxidants (diphenylene iodonium and apocynin) and gp91phox siRNA. In conclusion, oxLDL has concentration-dependent biphasic effects on human late-outgrowth EPC tube formation in vitro and in vivo.

No MeSH data available.


Related in: MedlinePlus