Hyperhomocyst(e)inemia impairs angiogenesis in a murine model of limb ischemia

Marta Bosch-Marcé, Roberto Pola, Andrea B. Wecker, Marcy Silver, Alberto Weber, Corinne Luedemann, Cynthia Curry, Toshinori Murayama, Marianne Kearney, Young Sup Yoon, M. René Malinow, Takayuki Asahara, Jeffrey M. Isner, Douglas W. Losordo

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Abstract

Hyperhomocyst(e)inemia (HH) is an established independent risk factor for coronary, cerebral and peripheral vascular diseases. Recent studies have indicated that certain cardiovascular risk factors, including diabetes and hypercholesterolemia, impair expression of vascular endothelial growth factor (VEGF) and endogenous angiogenesis. In this study, we investigate the impact of moderate HH on angiogenesis and VEGF pathway in a mouse model of hindlimb ischemia. Upon induction of unilateral hindlimb ischemia, endogenous angiogenesis, expression of VEGF, and phosphorylation of the VEGF receptor Flk-1 were evaluated in mice heterozygous for a deletion of the cystathionine β-synthase gene (CBS) and compared with those observed in CBS+/+ mice. CBS+/- mice exhibit moderate HH, as demonstrated by measuring plasma total homocyst(e)ine (tHcy) levels, which were significantly higher in these animals compared with CBS+/+ mice (4.77 ± 0.82 vs 2.10 ± 0.28, p < 0.01). Twenty-eight days after induction of ischemia, hindlimb blood flow was significantly reduced in CBS+/- mice compared with CBS+/+ animals (0.49 ± 0.03, n = 12 vs 0.71 ± 0.09, n = 10; P < 0.05). In addition, there was a significant negative correlation between plasma homocyst(e)ine levels and the laser Doppler perfusion ratio in CBS+/- mice (p = 0.0087, r = -0.7171). While VEGF expression and Flk-1 phosphorylation were not impaired in the ischemic muscles of CBS+/- mice, phosphorylation of the endothelial cell survival factor Akt was significantly inhibited by homocyst(e)ine in a dose-dependent manner in human umbilical vein endothelial cell (HUVECs) in vitro. In conclusion, our findings demonstrate that endogenous angiogenesis is inversely related to plasma levels of homocyst(e)ine in genetically engineered, heterozygous mice with moderate HH. This impairment, however, is not dependent on reduced expression of VEGF or impaired phosphorylation of its receptor Flk-1. In contrast, our data suggest that impaired Akt phosphorylation mediates the impairment of angiogenesis associated with HH.

Original languageEnglish
Pages (from-to)15-22
Number of pages8
JournalVascular Medicine
Volume10
Issue number1
DOIs
Publication statusPublished - 2005 May 26

Fingerprint

Ischemia
Extremities
Vascular Endothelial Growth Factor A
Phosphorylation
Hindlimb
Cystathionine
Vascular Endothelial Growth Factor Receptor-1
Peripheral Vascular Diseases
Human Umbilical Vein Endothelial Cells
Hypercholesterolemia
Cell Survival
Lasers
Endothelial Cells
Perfusion
Muscles
Genes

All Science Journal Classification (ASJC) codes

  • Cardiology and Cardiovascular Medicine

Cite this

Bosch-Marcé, M., Pola, R., Wecker, A. B., Silver, M., Weber, A., Luedemann, C., ... Losordo, D. W. (2005). Hyperhomocyst(e)inemia impairs angiogenesis in a murine model of limb ischemia. Vascular Medicine, 10(1), 15-22. https://doi.org/10.1191/1358863x05vm585oa
Bosch-Marcé, Marta ; Pola, Roberto ; Wecker, Andrea B. ; Silver, Marcy ; Weber, Alberto ; Luedemann, Corinne ; Curry, Cynthia ; Murayama, Toshinori ; Kearney, Marianne ; Yoon, Young Sup ; Malinow, M. René ; Asahara, Takayuki ; Isner, Jeffrey M. ; Losordo, Douglas W. / Hyperhomocyst(e)inemia impairs angiogenesis in a murine model of limb ischemia. In: Vascular Medicine. 2005 ; Vol. 10, No. 1. pp. 15-22.
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Bosch-Marcé, M, Pola, R, Wecker, AB, Silver, M, Weber, A, Luedemann, C, Curry, C, Murayama, T, Kearney, M, Yoon, YS, Malinow, MR, Asahara, T, Isner, JM & Losordo, DW 2005, 'Hyperhomocyst(e)inemia impairs angiogenesis in a murine model of limb ischemia', Vascular Medicine, vol. 10, no. 1, pp. 15-22. https://doi.org/10.1191/1358863x05vm585oa

Hyperhomocyst(e)inemia impairs angiogenesis in a murine model of limb ischemia. / Bosch-Marcé, Marta; Pola, Roberto; Wecker, Andrea B.; Silver, Marcy; Weber, Alberto; Luedemann, Corinne; Curry, Cynthia; Murayama, Toshinori; Kearney, Marianne; Yoon, Young Sup; Malinow, M. René; Asahara, Takayuki; Isner, Jeffrey M.; Losordo, Douglas W.

In: Vascular Medicine, Vol. 10, No. 1, 26.05.2005, p. 15-22.

Research output: Contribution to journalArticle

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T1 - Hyperhomocyst(e)inemia impairs angiogenesis in a murine model of limb ischemia

AU - Bosch-Marcé, Marta

AU - Pola, Roberto

AU - Wecker, Andrea B.

AU - Silver, Marcy

AU - Weber, Alberto

AU - Luedemann, Corinne

AU - Curry, Cynthia

AU - Murayama, Toshinori

AU - Kearney, Marianne

AU - Yoon, Young Sup

AU - Malinow, M. René

AU - Asahara, Takayuki

AU - Isner, Jeffrey M.

AU - Losordo, Douglas W.

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N2 - Hyperhomocyst(e)inemia (HH) is an established independent risk factor for coronary, cerebral and peripheral vascular diseases. Recent studies have indicated that certain cardiovascular risk factors, including diabetes and hypercholesterolemia, impair expression of vascular endothelial growth factor (VEGF) and endogenous angiogenesis. In this study, we investigate the impact of moderate HH on angiogenesis and VEGF pathway in a mouse model of hindlimb ischemia. Upon induction of unilateral hindlimb ischemia, endogenous angiogenesis, expression of VEGF, and phosphorylation of the VEGF receptor Flk-1 were evaluated in mice heterozygous for a deletion of the cystathionine β-synthase gene (CBS) and compared with those observed in CBS+/+ mice. CBS+/- mice exhibit moderate HH, as demonstrated by measuring plasma total homocyst(e)ine (tHcy) levels, which were significantly higher in these animals compared with CBS+/+ mice (4.77 ± 0.82 vs 2.10 ± 0.28, p < 0.01). Twenty-eight days after induction of ischemia, hindlimb blood flow was significantly reduced in CBS+/- mice compared with CBS+/+ animals (0.49 ± 0.03, n = 12 vs 0.71 ± 0.09, n = 10; P < 0.05). In addition, there was a significant negative correlation between plasma homocyst(e)ine levels and the laser Doppler perfusion ratio in CBS+/- mice (p = 0.0087, r = -0.7171). While VEGF expression and Flk-1 phosphorylation were not impaired in the ischemic muscles of CBS+/- mice, phosphorylation of the endothelial cell survival factor Akt was significantly inhibited by homocyst(e)ine in a dose-dependent manner in human umbilical vein endothelial cell (HUVECs) in vitro. In conclusion, our findings demonstrate that endogenous angiogenesis is inversely related to plasma levels of homocyst(e)ine in genetically engineered, heterozygous mice with moderate HH. This impairment, however, is not dependent on reduced expression of VEGF or impaired phosphorylation of its receptor Flk-1. In contrast, our data suggest that impaired Akt phosphorylation mediates the impairment of angiogenesis associated with HH.

AB - Hyperhomocyst(e)inemia (HH) is an established independent risk factor for coronary, cerebral and peripheral vascular diseases. Recent studies have indicated that certain cardiovascular risk factors, including diabetes and hypercholesterolemia, impair expression of vascular endothelial growth factor (VEGF) and endogenous angiogenesis. In this study, we investigate the impact of moderate HH on angiogenesis and VEGF pathway in a mouse model of hindlimb ischemia. Upon induction of unilateral hindlimb ischemia, endogenous angiogenesis, expression of VEGF, and phosphorylation of the VEGF receptor Flk-1 were evaluated in mice heterozygous for a deletion of the cystathionine β-synthase gene (CBS) and compared with those observed in CBS+/+ mice. CBS+/- mice exhibit moderate HH, as demonstrated by measuring plasma total homocyst(e)ine (tHcy) levels, which were significantly higher in these animals compared with CBS+/+ mice (4.77 ± 0.82 vs 2.10 ± 0.28, p < 0.01). Twenty-eight days after induction of ischemia, hindlimb blood flow was significantly reduced in CBS+/- mice compared with CBS+/+ animals (0.49 ± 0.03, n = 12 vs 0.71 ± 0.09, n = 10; P < 0.05). In addition, there was a significant negative correlation between plasma homocyst(e)ine levels and the laser Doppler perfusion ratio in CBS+/- mice (p = 0.0087, r = -0.7171). While VEGF expression and Flk-1 phosphorylation were not impaired in the ischemic muscles of CBS+/- mice, phosphorylation of the endothelial cell survival factor Akt was significantly inhibited by homocyst(e)ine in a dose-dependent manner in human umbilical vein endothelial cell (HUVECs) in vitro. In conclusion, our findings demonstrate that endogenous angiogenesis is inversely related to plasma levels of homocyst(e)ine in genetically engineered, heterozygous mice with moderate HH. This impairment, however, is not dependent on reduced expression of VEGF or impaired phosphorylation of its receptor Flk-1. In contrast, our data suggest that impaired Akt phosphorylation mediates the impairment of angiogenesis associated with HH.

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Bosch-Marcé M, Pola R, Wecker AB, Silver M, Weber A, Luedemann C et al. Hyperhomocyst(e)inemia impairs angiogenesis in a murine model of limb ischemia. Vascular Medicine. 2005 May 26;10(1):15-22. https://doi.org/10.1191/1358863x05vm585oa