Inhibition of inflammatory endothelial responses by a pathway involving caspase activation and p65 cleavage

Biochemistry. 2001 Apr 17;40(15):4686-92. doi: 10.1021/bi002498n.

Abstract

Suppression of NF kappa B activation has been involved in the elimination of survival programs during endothelial cell (EC) apoptosis. We used alpha-tocopheryl succinate (alpha-TOS) to trigger apoptosome formation and the subsequent activation of executioner caspases. The level of bcl-2 was reduced by alpha-TOS, and its downregulation potentiated and its overexpression suppressed pro-apoptotic effects of alpha-TOS, indicating a mitochondrial role in alpha-TOS-induced apoptosis in EC. alpha-TOS treatment was associated with induction of TUNEL-positive apoptosis in EC with a high but not with a low proliferation index. The use of the pan-caspase inhibitor z-VAD.fmk suggested the involvement of caspases in cleavage of p65, and in inhibition of nuclear translocation of p65 and NF kappa B-dependent transactivation of a gene construct encoding the green fluorescence protein elicited by TNF alpha in contact-arrested EC. The suppression by alpha-TOS of inflammatory EC responses induced by TNF alpha such as VCAM-1 mRNA and surface protein expression and shear-resistant arrest of monocytic cells were also reversed by z-VAD.fmk. NF kappa B-dependent transactivation was preserved in alpha-TOS-treated EC stably transfected with a caspase-noncleavable p65 mutant but not with its truncated form, thus establishing a direct link between alpha-TOS-induced effects and p65 cleavage. Our data infer a pathway by which caspase activation in EC inhibits NF kappa B-dependent inflammatory activation and monocyte recruitment, and provide evidence for a relationship between pro-apoptotic and anti-inflammatory pathways.

MeSH terms

  • Apoptosis / drug effects
  • Caspase 3
  • Caspase 6
  • Caspases / metabolism*
  • Caspases / physiology
  • Cell Adhesion / drug effects
  • Cell Division / drug effects
  • Cell Line
  • Cells, Cultured
  • Down-Regulation
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / enzymology*
  • Endothelium, Vascular / metabolism
  • Endothelium, Vascular / pathology*
  • Enzyme Activation / drug effects
  • Humans
  • Hydrolysis / drug effects
  • Inflammation / enzymology
  • Inflammation / metabolism
  • Monocytes / drug effects
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / metabolism*
  • Signal Transduction / drug effects
  • Tocopherols
  • Transcription Factor RelA
  • Transcriptional Activation / drug effects
  • Vascular Cell Adhesion Molecule-1 / biosynthesis
  • Vascular Cell Adhesion Molecule-1 / genetics
  • Vitamin E / analogs & derivatives*
  • Vitamin E / pharmacology

Substances

  • NF-kappa B
  • Transcription Factor RelA
  • Vascular Cell Adhesion Molecule-1
  • Vitamin E
  • CASP3 protein, human
  • CASP6 protein, human
  • Caspase 3
  • Caspase 6
  • Caspases
  • Tocopherols