Abstract | BACKGROUND: METHODS: H2S production from recombinant human CBS, CSE and 3-MST was measured using the fluorescent H2S probe AzMC. Mouse liver homogenate (a rich source of CBS) was also employed to measure H2S biosynthesis. The interaction of copper with accessible protein cysteine residues was evaluated using the DTNB method. Cell proliferation and viability were measured using the BrdU and MTT methods. Cellular bioenergetics was evaluated by Extracellular Flux Analysis. RESULTS: While disulfiram did not exert any significant direct inhibitory effect on any of the H2S-producing enzymes, its metabolite, CuDDC was a potent inhibitor of CBS and CSE. The mode of its action is likely related to the complexed copper molecule. In cell-based systems, the effects of disulfiram were variable. In colon cancer cells, no significant effect of disulfiram was observed on H2S production or proliferation or viability. In contrast, in DS fibroblasts, disulfiram inhibited H2S production and improved proliferation and viability. Copper, on its own, failed to have any effects on either cell type, likely due to its low cell penetration. CuDDC inhibited H2S production in both cell types studied and exerted the functional effects that would be expected from a CBS inhibitor: inhibition of cell proliferation of cancer cells and a bell-shaped effect (stimulation of proliferation at low concentration and inhibition of these responses at higher concentration) in DS cells. Control experiments using a chemical H2S donor showed that, in addition to inhibiting CBS and CSE, part of the biological effects of CuDDC relates to a direct reaction with H2S, which occurs through its complexed copper. CONCLUSIONS:
Disulfiram, via its metabolite CuDDC acts as an inhibitor of CBS and a scavenger of H2S, which, in turn, potently suppresses H2S levels in various cell types. Inhibition of H2S biosynthesis may explain some of the previously reported actions of disulfiram and CuDDC in vitro and in vivo. Disulfiram or CuDDC may be considered as potential agents for the experimental therapy of various pathophysiological conditions associated with H2S overproduction.
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Authors | Karim Zuhra, Theodora Panagaki, Elisa B Randi, Fiona Augsburger, Marc Blondel, Gaelle Friocourt, Yann Herault, Csaba Szabo |
Journal | Biochemical pharmacology
(Biochem Pharmacol)
Vol. 182
Pg. 114267
(12 2020)
ISSN: 1873-2968 [Electronic] England |
PMID | 33035509
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved. |
Chemical References |
- Acetaldehyde Dehydrogenase Inhibitors
- Chelating Agents
- Organometallic Compounds
- bis(N,N-diethyldithiocarbamate)Cu (II) complex
- Copper
- Ditiocarb
- Cystathionine beta-Synthase
- Disulfiram
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Topics |
- Acetaldehyde Dehydrogenase Inhibitors
(metabolism, pharmacology)
- Animals
- Cell Survival
(drug effects, physiology)
- Chelating Agents
(metabolism, pharmacology)
- Copper
(metabolism, pharmacology)
- Cystathionine beta-Synthase
(antagonists & inhibitors, metabolism)
- Disulfiram
(metabolism, pharmacology)
- Ditiocarb
(analogs & derivatives, metabolism, pharmacology)
- Dose-Response Relationship, Drug
- Female
- HCT116 Cells
- Humans
- Liver
(drug effects, metabolism)
- Mice
- Mice, Inbred BALB C
- Organometallic Compounds
(metabolism, pharmacology)
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