Ratiometric fluorophores to ONOO– in living cells and monitoring endothelial dysfunction

초록

Peroxynitrite (ONOO–), generated from nitric oxide and superoxide, is implicated in cellular dysfunction and death. To detect ONOO– in live cells, we developed two hemicyanine-based ratiometric fluorophores, P-mito and P-boc. These ratiometric fluorophores were successfully used to monitor ONOO– fluctuations in cancer cells and endothelial cells. P-mito predominantly accumulated in the mitochondria of live cells and, upon selective reaction with ONOO–, increased the ratio of the fluorescence intensity from blue channel to that from red channel. Treatment with an ONOO– scavenger, N-acetyl cysteine, suppressed this ratio increase. Notably, in a model of endothelial dysfunction induced by treating pulmonary arterial endothelial cells with TGFβ2 and IL-1β, our system successful demonstrated increased ONOO– levels, effectively highlighting the utility of the hemicyanine-based fluorescence response to ONOO–. Therefore, our hemicyanine-based ONOO– detection system provides an effective platform for spatiotemporal monitoring and quantitative analysis of ONOO– within live cells, enabling molecular-level research into various ONOO–-related pathological conditions. © 2024 Elsevier B.V.

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