Copper diethyldithiocarbamate as an inhibitor of tissue plasminogen activator synthesis in cultured human coronary endothelial cells

Abstract

-Recent developments have shown that organic-inorganic hybrid molecules have the potential to provide useful tools for analyzing biological systems. In the case of fibrinolysis, which is the phenomenon whereby fibrin is degraded by plasmin that has been converted from plasminogen via tissue plasminogen activator (t-PA) secreted from vascular endothelial cells, we hypothesized that there may be organic-inorganic hybrid molecules that could be used to analyze the mechanisms by which endothelial fibrinolysis is regulated. In our present study, we found that a copper complex -copper diethyldithiocarbamate (Cu10) -reduces t-PA activity in a conditioned medium of cultured human coronary endothelial cells by inhibiting the t-PA synthesis without changing the synthesis of plasminogen activator inhibitor type 1, which is a t-PA inhibitor. Copper sulfate, the Cu10 ligand, and zinc/iron complexes with the same Cu10 ligand, did not exhibit such biological activity. These results indicate that Cu10 has the potential to provide a useful tool for finding alternative pathways that downregulate endothelial t-PA synthesis.

Nuclear factor erythroid 2-related factor 2 (NRF2) is a negative regulator of tissue plasminogen activator synthesis in cultured human vascular endothelial EA.hy926 cells

“…Meanwhile, our previous experiments showed that cAMP, which down-regulates t-PA synthe-sis, was not involved in endothelin-1's inhibitory effect on endothelial t-PA synthesis in human umbilical vein endothelial cells (Kaji et al, 1992a). Recently, we also reported that copper diethyldithiocarbamate down-regulates endothelial t-PA synthesis in human coronary endothelial cells but the cAMP pathway is not involved in the inhibitory effect (Fujie et al, 2017). Because copper diethyldithiocarbamate is an activator of NRF2 in vascular endothelial cells (Fujie et al, 2016), the compound likely activates the NRF2 pathway and then inhibits the t-PA synthesis in human coronary endothelial cells and EA.hy926 cells.…”

“…t-PA and PAI-1 secretion from vascular endothelial cells is influenced by not only various endogenous fac-tors including thrombin (Levin et al, 1984;Gelehrter and Sznycer-Laszuk, 1986), endothelin (Kaji et al, 1992a;Yamamoto et al, 1992Yamamoto et al, , 1993a, cytokines (Schleef et al, 1988) and growth factors (Kaji et al, 1994; but also heavy metals such as lead (Kaji et al, 1992b) and cadmium (Yamamoto et al, 1993b). Recently, we have also shown that copper diethyldithiocarbamate, which is a copper complex that exhibits interesting biological activities including activation of nuclear factor erythroid 2-related factor 2 (NRF2) (Fujie et al, 2016), suppresses the fibrinolytic activity of human coronary endothelial cells by down-regulating t-PA synthesis (Fujie et al, 2017). Although transcription factor NRF2 mainly regulates the gene expression of antioxidant and phase II xenobiotic metabolizing enzymes such as NAD(P)H quinone oxidoreductase 1 (NQO1) and heme oxygenase-1 (HO-1) (Kensler et al, 2007), the role of NRF2 in regulating endothelial fibrinolytic activity remains unclear.…”

A zinc complex that suppresses the expression of a reactive sulfur species-producing enzyme, cystathionine γ-lyase, in cultured vascular endothelial cells

“…Organic-inorganic hybrid molecules used in these reports exhibited selective biological activities. For example, copper(II) bis(diethyldithiocarbamate), which activates the transcription factor NF-E2-related factor 2 (Nrf2) (Fujie et al, 2016d) and modulates the fibrinolytic activity (Fujie et al, 2017) in vascular endothelial cells, induces the expression of only syndecan-4 among the several types of endothelial proteoglycans (Hara et al, 2018). The selective suppression of endothelial CSE expression by Zn11 suggests the suitability of this zinc complex as a molecular probe for the analysis of the mechanisms underlying endothelial CSE expression.…”

Copper in the suprachiasmatic circadian clock: A possible link between multiple circadian oscillators

“…Changes in cellular redox state in turn can influence membrane excitability, such as through modulating K + currents. Cu has been shown to interact with extracellular and intracellular signal transduction mechanisms, including modulating NMDA, MAPK, and TrkB signaling, a variety of ion channels, and extracellular matrix/proteases (e.g., collagen lyase, PAI‐1), all of which again can alter membrane excitability (Bucci et al., , ; D'Ambrosi & Rossi, ; Fujie et al., ; Migocka, ; Scheiber et al., ; Urso & Maffia, ; Zlatic et al., ). Our data are consistent with this, showing Cu/Cu depletion‐induced effects involving NMDA, MAPK, TrkB, and NO.…”