Summary
The addition of thrombin (9 nM) to primary cultures of human endothelial cells induces
a 6- to 7-fold increase in the rate of release of tissue plasminogen activator (tPA).
Several other serine proteases which specifically interact with endothelial cells
were also analyzed for their effect on tPA release. Gamma-thrombin, an autocatalytic
product of α-thrombin, promoted tPA release but was less effective than α-thrombin.
A maximum increase of 5.5-fold was observed, although a concentration of γ-thrombin
20 times greater than α-thrombin was required. The response to Factor Xa was similar to α-thrombin, although the stimulation was significantly reduced by
the addition of hirudin or DAPA suggesting that prothrombin activation was occurring.
The simultaneous addition of prothrombin with Factor Xa resulted in enhanced tPA release equal to that observed with an equimolar concentration
of active α-thrombin. Thus, under these conditions, Factor Xa-cell surface mediated activation of prothrombin can lead to a secondary effect resulting
from cell-thrombin interaction. Activated protein C, which has been implicated as
a profibrinolytic agent, was also tested. No change in tPA release occurred after
the addition of up to 325 nM activated protein C in the presence or absence of proteins.
Factor IXa and plasmin were also ineffective. The effect of thrombin on the endothelial cell
derived plasminogen activator specific inhibitor was also studied. Thrombin produced
a small but variable release of the inhibitor with an increase of less than twice
that of non-thrombin treated controls.
Key words
Thrombin - Tissue plasminogen activator - Endothelial cells - Plasminogen activator
specific inhibitor
