p21Waf1/Cip1/Sdi1 mediates shear stress-dependent antiapoptotic function

Stefania Mattiussi, Paolo Turrini, Lucia Testolin, Fabio Martelli, Germana Zaccagnini, Antonella Mangoni, Laura M. Barlucchi, Annalisa Antonini, Barbara Illi, Corrado Cirielli, Julio Padron, Chiara Nicolò, Roberto Testi, Francesco Osculati, Paolo Biglioli, Maurizio C. Capogrossi, Carlo Gaetano

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Objective: The antiapoptotic effect of p21Waf1/Cip1/Sdi1 (p21) was examined in human umbilical vein endothelial cells (HUVEC) exposed to laminar shear stress (SS) or to the nitric oxide donor sodium nitroprusside (SNP) and in a mouse model of hindlimb ischemia. Methods: In vitro: Cells were cultured without serum and in the presence of cobalt chloride to simulate hypoxia for 12 h (T0). Shear stress was applied to endothelial cells for additional 12 h. In vivo: Hindlimb ischemia was realized in mice by femoral artery ligation. SNP was acutely administered by subcutaneous injection or by Alzet osmotic pumps for a longer treatment. Results: At T0, HUVEC were either exposed to SS (15 dyn/cm2/s-1), treated with SNP or kept in static condition (ST) for 1-12 h; after additional 12 h in ST, 30-35% of cells still alive at T0 had died. In this condition, both SS and SNP treatments markedly increased p21 levels and reduced apoptosis in HUVEC. Recombinant adenoviruses carrying p21 (AdCMV.p21) or antisense p21 (AdCMV.ASp21) cDNA revealed that AdCMV.p21-infected HUVEC were protected from death while AdCMV.ASp21 reduced SS- and SNP-dependent protection from apoptosis. In mice, apoptosis was detected in endothelial cells of ischemic hindlimbs as early as 8 h after femoral artery ligation. Treatment with SNP enhanced p21 expression and protected ischemic tissue from damage. Remarkably, direct in vivo injection of AdCMV.p21 significantly reduced the number of apoptotic nuclei in the presence of ischemia. Conclusions: The present study establishes that, under our experimental conditions, (a) p21 plays an important role in SS and nitric oxide antiapoptotic effect in vitro, and (b) p21 gene transfer prevents apoptosis in vitro and in vivo, following acute interruption of blood flow.

Original languageEnglish
Pages (from-to)693-704
Number of pages12
JournalCardiovascular Research
Volume61
Issue number4
DOIs
Publication statusPublished - Mar 1 2004

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Nitroprusside
Human Umbilical Vein Endothelial Cells
Hindlimb
Apoptosis
Ischemia
Femoral Artery
Ligation
Endothelial Cells
Nitric Oxide Donors
Subcutaneous Injections
Adenoviridae
Cultured Cells
Nitric Oxide
Therapeutics
Complementary DNA
Injections
Serum
Genes
In Vitro Techniques

Keywords

  • Apoptosis
  • Flow
  • Hypoxia
  • Ischemia
  • p21
  • Shear stress

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

p21Waf1/Cip1/Sdi1 mediates shear stress-dependent antiapoptotic function. / Mattiussi, Stefania; Turrini, Paolo; Testolin, Lucia; Martelli, Fabio; Zaccagnini, Germana; Mangoni, Antonella; Barlucchi, Laura M.; Antonini, Annalisa; Illi, Barbara; Cirielli, Corrado; Padron, Julio; Nicolò, Chiara; Testi, Roberto; Osculati, Francesco; Biglioli, Paolo; Capogrossi, Maurizio C.; Gaetano, Carlo.

In: Cardiovascular Research, Vol. 61, No. 4, 01.03.2004, p. 693-704.

Research output: Contribution to journalArticle

Mattiussi, S, Turrini, P, Testolin, L, Martelli, F, Zaccagnini, G, Mangoni, A, Barlucchi, LM, Antonini, A, Illi, B, Cirielli, C, Padron, J, Nicolò, C, Testi, R, Osculati, F, Biglioli, P, Capogrossi, MC & Gaetano, C 2004, 'p21Waf1/Cip1/Sdi1 mediates shear stress-dependent antiapoptotic function', Cardiovascular Research, vol. 61, no. 4, pp. 693-704. https://doi.org/10.1016/j.cardiores.2003.12.008
Mattiussi, Stefania ; Turrini, Paolo ; Testolin, Lucia ; Martelli, Fabio ; Zaccagnini, Germana ; Mangoni, Antonella ; Barlucchi, Laura M. ; Antonini, Annalisa ; Illi, Barbara ; Cirielli, Corrado ; Padron, Julio ; Nicolò, Chiara ; Testi, Roberto ; Osculati, Francesco ; Biglioli, Paolo ; Capogrossi, Maurizio C. ; Gaetano, Carlo. / p21Waf1/Cip1/Sdi1 mediates shear stress-dependent antiapoptotic function. In: Cardiovascular Research. 2004 ; Vol. 61, No. 4. pp. 693-704.
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AU - Turrini, Paolo

AU - Testolin, Lucia

AU - Martelli, Fabio

AU - Zaccagnini, Germana

AU - Mangoni, Antonella

AU - Barlucchi, Laura M.

AU - Antonini, Annalisa

AU - Illi, Barbara

AU - Cirielli, Corrado

AU - Padron, Julio

AU - Nicolò, Chiara

AU - Testi, Roberto

AU - Osculati, Francesco

AU - Biglioli, Paolo

AU - Capogrossi, Maurizio C.

AU - Gaetano, Carlo

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N2 - Objective: The antiapoptotic effect of p21Waf1/Cip1/Sdi1 (p21) was examined in human umbilical vein endothelial cells (HUVEC) exposed to laminar shear stress (SS) or to the nitric oxide donor sodium nitroprusside (SNP) and in a mouse model of hindlimb ischemia. Methods: In vitro: Cells were cultured without serum and in the presence of cobalt chloride to simulate hypoxia for 12 h (T0). Shear stress was applied to endothelial cells for additional 12 h. In vivo: Hindlimb ischemia was realized in mice by femoral artery ligation. SNP was acutely administered by subcutaneous injection or by Alzet osmotic pumps for a longer treatment. Results: At T0, HUVEC were either exposed to SS (15 dyn/cm2/s-1), treated with SNP or kept in static condition (ST) for 1-12 h; after additional 12 h in ST, 30-35% of cells still alive at T0 had died. In this condition, both SS and SNP treatments markedly increased p21 levels and reduced apoptosis in HUVEC. Recombinant adenoviruses carrying p21 (AdCMV.p21) or antisense p21 (AdCMV.ASp21) cDNA revealed that AdCMV.p21-infected HUVEC were protected from death while AdCMV.ASp21 reduced SS- and SNP-dependent protection from apoptosis. In mice, apoptosis was detected in endothelial cells of ischemic hindlimbs as early as 8 h after femoral artery ligation. Treatment with SNP enhanced p21 expression and protected ischemic tissue from damage. Remarkably, direct in vivo injection of AdCMV.p21 significantly reduced the number of apoptotic nuclei in the presence of ischemia. Conclusions: The present study establishes that, under our experimental conditions, (a) p21 plays an important role in SS and nitric oxide antiapoptotic effect in vitro, and (b) p21 gene transfer prevents apoptosis in vitro and in vivo, following acute interruption of blood flow.

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