A1 functions at the mitochondria to delay endothelial apoptosis in response to tumor necrosis factor

Tumor necrosis factor (TNF) does not cause endothelial apoptosis unless the expression of cytoprotective genes is blocked. We have previously demonstrated that one of the TNF-inducible cytoprotective genes is the Bcl-2 family member, Al. Al is induced by the action of the transcription factor, NF kappa B, in response to inflammatory mediators. In this report we demonstrate that, as with other cell types, inhibition of NF kappa B initiates microvascular endothelial apoptosis in response to TNF. Al is able to inhibit this apoptosis over 24 h, We demonstrate that Al is localized to and functions at the mitochondria. Whereas Al is able to inhibit mitochondrial depolarization, loss of cytochrome c, cleavage of caspase 9, BID, and poly(ADP-ribose) polymerase, it does not block caspase 8 or caspase 3 cleavage. In contrast, Al is not able to prevent endothelial apoptosis by TNF over 72 h, when NF kappa B signaling is blocked. On the other hand, the caspase inhibitor, benzyloxycarbonyl-VAD-formylmethyl ketone, completely blocks TNF-induced endothelial apoptosis over 72 h. Our findings indicate that Al is able to maintain temporary survival of endothelial cells in response to TNF by maintaining mitochondrial viability and function. However, a mitochondria-independent caspase pathway eventually results in endothelial death despite mitochondrial protection by A1.