17O NMR spectroscopy-assisted in vitro bioactivity studies of the intermediates formed via Na2S and RSNO cross-linking reactions

RSC Advances Pub Date: 2020-10-29 DOI: 10.1039/D0RA05054G

Abstract

The cross-linking reaction between sulfide and S-nitrosothiol moieties has been intensively investigated and thionitrite/thionitrous acid (SNO?/HSNO) as well as nitrosopersulfide (SSNO?) were reported to be the intermediates that could serve as reservoirs for nitric oxide (NO). However, debate still exists regarding the stability and biological activity of SNO?/HSNO and SSNO?. In order to investigate the chemical properties and biological activity of SNO? and SSNO?, we set out to re-characterize the reaction intermediates using UV-Vis and 15N NMR spectroscopy techniques, as well as a new 17O NMR approach. The effects of SNO? and SSNO? on cellular NO and cGMP levels were assessed via cell culture experiments, and also the effects of SNO? and SSNO? on cell proliferation, migration, and capillary-like structure formation were evaluated with human umbilical vein endothelial cells (HUVEC). Through this work, the characteristic peaks and half-lives of SNO? and SSNO? were elucidated under various preparation conditions. The biological assays demonstrated that SSNO? increased the cellular NO and cGMP levels and also facilitated cell proliferation, migration and stimulated angiogenesis, while in contrast SNO? did not exhibit these effects.

Graphical abstract: 17O NMR spectroscopy-assisted in vitro bioactivity studies of the intermediates formed via Na2S and RSNO cross-linking reactions
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