p21(Waf1/Cip1/Sdi1) mediates shear stress-dependent antiapoptotic function. Export

@article{citeulike:3361500, abstract = {OBJECTIVE: The antiapoptotic effect of p21(Waf1/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.}, author = {Mattiussi, S. and Turrini, P. and Testolin, L. and Martelli, F. and Zaccagnini, G. and Mangoni, A. and Barlucchi, L. M. and Antonini, A. and Illi, B. and Cirielli, C. and Padron, J. and Nicol\`{o}, C. and Testi, R. and Osculati, F. and Biglioli, P. and Capogrossi, M. C. and Gaetano, C.}, citeulike-article-id = {3361500}, citeulike-linkout-0 = {http://dx.doi.org/10.1016/j.cardiores.2003.12.008}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/14985066}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=14985066}, day = {1}, doi = {10.1016/j.cardiores.2003.12.008}, issn = {0008-6363}, journal = {Cardiovascular research}, keywords = {apoptosis, cobalt, huvec, hypoxia, in\_vivo, ischemia, nitric\_oxide, no, p21, shear, shear\_stress}, month = {March}, number = {4}, pages = {693--704}, posted-at = {2008-10-01 07:46:23}, priority = {4}, title = {p21(Waf1/Cip1/Sdi1) mediates shear stress-dependent antiapoptotic function.}, url = {http://dx.doi.org/10.1016/j.cardiores.2003.12.008}, volume = {61}, year = {2004} }

TY - JOUR ID - citeulike:3361500 L3 - citeulike-article-id:3361500 N2 - OBJECTIVE: The antiapoptotic effect of p21(Waf1/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. IS - 4 JF - Cardiovascular research SN - 0008-6363 EP - 704 TI - p21(Waf1/Cip1/Sdi1) mediates shear stress-dependent antiapoptotic function. VL - 61 SP - 693 KW - apoptosis KW - cobalt KW - huvec KW - hypoxia KW - in_vivo KW - ischemia KW - nitric_oxide KW - no KW - p21 KW - shear KW - shear_stress AU - Mattiussi, S AU - Turrini, P AU - Testolin, L AU - Martelli, F AU - Zaccagnini, G AU - Mangoni, A AU - Barlucchi, LM AU - Antonini, A AU - Illi, B AU - Cirielli, C AU - Padron, J AU - Nicolò, C AU - Testi, R AU - Osculati, F AU - Biglioli, P AU - Capogrossi, MC AU - Gaetano, C PY - 2004/03/01/ UR - http://dx.doi.org/10.1016/j.cardiores.2003.12.008 ER -