15-Deoxy-Δ(12,14)-prostaglandin J₂ induces vascular endothelial cell apoptosis through the sequential activation of MAPKS and p53

15-Deoxy-Δ(12,14)-prostaglandin J₂ (15d-PGJ₂) is a potent anti-angiogenic factor and induces endothelial cell apoptosis, although the mechanism remains unclear. In this study, 15d-PGJ₂ was found to increase p53 levels of the human umbilical vein endothelial cells by stabilizing p53. Both 15d-PGJ₂-induced apoptosis and the induction of p21^Waf1 and Bax can be abolished by p53 small interfering RNA but not by peroxisome proliferator-activated receptor γ inhibitors. Moreover, 15d-PGJ₂ activated JNK and p38 MAPK while inducing p53 phosphorylation at sites responsible for p53 activity. JNK inhibitor (SP600125) or p38 MAPK inhibitor (SB203580) pretreatment attenuated 15d-PGJ ₂-mediated apoptosis and suppressed the p21^Waf1 and Bax expressions without affecting p53 protein accumulation. Pretreatment with SP600125 partially prevented the phosphorylation of p53 at serines 33 and 392 induced by 15d-PGJ₂. 15d-PGJ₂ was also found to induce reactive oxygen species generation and partially blocked nuclear factor-κB activity. Pretreatment with antioxidant N-acetylcysteine prevented the p53 accumulation, the phosphorylations of JNK and p38 MAPK, the inhibition of NF-κB activity, as well as the apoptosis induced by 15d-PGJ₂. Using a mouse model of corneal neovascularization, it was demonstrated in vivo that 15d-PGJ₂ induced reactive oxygen species generation, activated JNK and p38 MAPK, induced p53 accumulation/phosphorylation, and induced vascular endothelial cell apoptosis, which could be abolished by N-acetylcysteine, SP600125, SB203580, or a virus-derived amphipathic peptides-based p53 small interfering RNA. This is the first study that 15d-PGJ₂ induces vascular endothelial cell apoptosis through the signaling of JNK and p38 MAPK-mediated p53 activation both in vitro and in vivo, further establishing the potential of 15d-PGJ₂ as an anti-angiogenesis agent.