Pyrrolidine dithiocarbamate activates the heat shock response and thereby induces apoptosis in primed endothelial cells.

Abstract

Transcription factor NF-kappaB is an important regulator of the cellular response to diverse stresses. Pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kappaB activity, was used to determine the role of this transcription factor in our model of stress-induced endothelial cell apoptosis. Porcine aortic endothelial cells were treated with an inducer of the acute phase response (LPS) followed by treatment with an inducer of the heat shock response (arsenite), a sequence that produces cell death by apoptosis. Treatment with PDTC attenuated LPS-induced NF-kappaB activity and endothelial cell death when added prior to LPS. However, PDTC unexpectedly increased cell death when given after LPS priming. This time-dependent effect of PDTC on endothelial cell death was similar to that which we had observed previously for inducers of the heat shock response. Therefore, we hypothesized that PDTC could induce the heat shock response in porcine and human endothelial cells. PDTC increased heat shock protein (HSP)-70 production and heat shock factor (HSF) activity. Thus, treatment of endothelial cells with PDTC, like other inducers of the heat shock response, increased HSP-70 levels and HSF activity and had time-dependent effects on cell death by apoptosis in primed endothelial cells. We conclude that PDTC induced the heat shock response, that induction of HSF activity may be linked with inhibition of NF-kappaB activity, and that interaction between acute phase and heat shock regulatory factors may be pivotal to determining cell fate (apoptosis).