Hypoxia-Induced Endothelial Apoptosis Through Nuclear Factor-kappaB (NF-kappaB)-Mediated bcl-2 Suppression : In Vivo Evidence of the Importance of NF-kappaB in Endothelial Cell Regulation Export

@article{matsushita2000, abstract = {Abstract--The transcription factor nuclear factor-{kappa}B (NF-{kappa}B) plays a pivotal role in the coordinated transactivation of cytokine and adhesion molecule genes involved in endothelial activation. Although recent reports have documented the contribution of NF-{kappa}B to apoptosis, it is still controversial. Especially, the role of NF-{kappa}B in endothelial apoptosis is largely unknown. Hypoxia significantly induced human aortic endothelial cell death and apoptosis in a time-dependent manner (P<0.01), accompanied by NF-{kappa}B activation. Decrease in total cell number and increase in apoptotic cells induced by hypoxia were significantly attenuated by NF-{kappa}B decoy, but not by scrambled decoy, oligodeoxynucleotides (ODNs) (P<0.01). Increase in DNA fragmentation induced by hypoxia was also significantly inhibited by NF-{kappa}B decoy ODNs as compared with scrambled decoy ODNs (P<0.01). Moreover, transfection of NF-{kappa}B decoy ODNs resulted in a significant decrease in caspase-3-like activity, which is a common pathway for apoptosis, compared with scrambled decoy ODNs. Importantly, transfection of NF-{kappa}B decoy ODNs significantly increased protein of bcl-2, an inhibitor of apoptosis, and did not alter bax, a promoter of apoptosis, thereby resulting in a significant increase in the ratio of bcl-2 to bax (P<0.01). bcl-2 mRNA was also decreased by hypoxia, whereas transfection of NF-{kappa}B decoy ODNs significantly attenuated decrease in bcl-2 mRNA. These results demonstrate that activation of NF-{kappa}B by hypoxia induced endothelial apoptosis in a bcl-2-dependent manner. The importance of NF-{kappa}B in endothelial apoptosis was confirmed by the observation that pyrrolidine dithiocarbamate, a potent NF-{kappa}B inhibitor, prevented endothelial apoptosis, caspase 3-like activity, and bcl-2 downregulation induced by hypoxia. To test this hypothesis in vivo, we transfected NF-{kappa}B decoy ODNs into rat intact carotid artery after reperfusion injury. Reperfusion injury was associated with a significant increase in endothelial apoptosis at 24 hours, whereas NF-{kappa}B decoy ODN treatment markedly decreased terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL)-positive endothelial cells at 24 hours after reperfusion (P<0.01). Here, using synthetic double-stranded DNA with high affinity for NF-{kappa}B as a decoy approach, we demonstrated that activation of NF-{kappa}B by hypoxia caused aortic endothelial cell death and apoptosis through the suppression of bcl-2. NF-{kappa}B-mediated endothelial apoptosis induced by hypoxia may be involved in the pathogenesis of endothelial dysfunction observed in cardiovascular ischemic diseases.}, author = {Matsushita, Hidetsugu and Morishita, Ryuichi and Nata, Toshie and Aoki, Motokuni and Nakagami, Hironori and Taniyama, Yoshiaki and Yamamoto, Kei and Higaki, Jitsuo and Yasufumi, Kaneda and Ogihara, Toshio}, citeulike-article-id = {2334839}, citeulike-linkout-0 = {http://circres.ahajournals.org/cgi/content/abstract/86/9/974}, eprint = {http://circres.ahajournals.org/cgi/reprint/86/9/974.pdf}, journal = {Circulation Research}, keywords = {endothelial, experimental, hypoxia, invivo, nfkb}, number = {9}, pages = {974--981}, pdf = {matsushita2000.pdf}, posted-at = {2008-02-05 13:12:53}, priority = {2}, title = {Hypoxia-Induced Endothelial Apoptosis Through Nuclear Factor-{kappa}B (NF-{kappa}B)-Mediated bcl-2 Suppression : In Vivo Evidence of the Importance of NF-{kappa}B in Endothelial Cell Regulation}, url = {http://circres.ahajournals.org/cgi/content/abstract/86/9/974}, volume = {86}, year = {2000} }

TY - JOUR ID - matsushita2000 L3 - citeulike-article-id:2334839 N2 - Abstract--The transcription factor nuclear factor-{kappa}B (NF-{kappa}B) plays a pivotal role in the coordinated transactivation of cytokine and adhesion molecule genes involved in endothelial activation. Although recent reports have documented the contribution of NF-{kappa}B to apoptosis, it is still controversial. Especially, the role of NF-{kappa}B in endothelial apoptosis is largely unknown. Hypoxia significantly induced human aortic endothelial cell death and apoptosis in a time-dependent manner (P<0.01), accompanied by NF-{kappa}B activation. Decrease in total cell number and increase in apoptotic cells induced by hypoxia were significantly attenuated by NF-{kappa}B decoy, but not by scrambled decoy, oligodeoxynucleotides (ODNs) (P<0.01). Increase in DNA fragmentation induced by hypoxia was also significantly inhibited by NF-{kappa}B decoy ODNs as compared with scrambled decoy ODNs (P<0.01). Moreover, transfection of NF-{kappa}B decoy ODNs resulted in a significant decrease in caspase-3-like activity, which is a common pathway for apoptosis, compared with scrambled decoy ODNs. Importantly, transfection of NF-{kappa}B decoy ODNs significantly increased protein of bcl-2, an inhibitor of apoptosis, and did not alter bax, a promoter of apoptosis, thereby resulting in a significant increase in the ratio of bcl-2 to bax (P<0.01). bcl-2 mRNA was also decreased by hypoxia, whereas transfection of NF-{kappa}B decoy ODNs significantly attenuated decrease in bcl-2 mRNA. These results demonstrate that activation of NF-{kappa}B by hypoxia induced endothelial apoptosis in a bcl-2-dependent manner. The importance of NF-{kappa}B in endothelial apoptosis was confirmed by the observation that pyrrolidine dithiocarbamate, a potent NF-{kappa}B inhibitor, prevented endothelial apoptosis, caspase 3-like activity, and bcl-2 downregulation induced by hypoxia. To test this hypothesis in vivo, we transfected NF-{kappa}B decoy ODNs into rat intact carotid artery after reperfusion injury. Reperfusion injury was associated with a significant increase in endothelial apoptosis at 24 hours, whereas NF-{kappa}B decoy ODN treatment markedly decreased terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL)-positive endothelial cells at 24 hours after reperfusion (P<0.01). Here, using synthetic double-stranded DNA with high affinity for NF-{kappa}B as a decoy approach, we demonstrated that activation of NF-{kappa}B by hypoxia caused aortic endothelial cell death and apoptosis through the suppression of bcl-2. NF-{kappa}B-mediated endothelial apoptosis induced by hypoxia may be involved in the pathogenesis of endothelial dysfunction observed in cardiovascular ischemic diseases. IS - 9 JF - Circulation Research EP - 981 TI - Hypoxia-Induced Endothelial Apoptosis Through Nuclear Factor-{kappa}B (NF-{kappa}B)-Mediated bcl-2 Suppression : In Vivo Evidence of the Importance of NF-{kappa}B in Endothelial Cell Regulation VL - 86 SP - 974 KW - endothelial KW - experimental KW - hypoxia KW - invivo KW - nfkb AU - Matsushita, Hidetsugu AU - Morishita, Ryuichi AU - Nata, Toshie AU - Aoki, Motokuni AU - Nakagami, Hironori AU - Taniyama, Yoshiaki AU - Yamamoto, Kei AU - Higaki, Jitsuo AU - Yasufumi, Kaneda AU - Ogihara, Toshio PY - 2000/// UR - http://circres.ahajournals.org/cgi/content/abstract/86/9/974 ER -