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Fluorescence probes to detect lipid-derived radicals

Nature Chemical Biology volume 12, pages 608–613 (2016)Cite this article

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Abstract

Lipids and their metabolites are easily oxidized in chain reactions initiated by lipid radicals, forming lipid peroxidation products that include the electrophiles 4-hydroxynonenal and malondialdehyde. These markers can bind cellular macromolecules, causing inflammation, apoptosis and other damage. Methods to detect and neutralize the initiating radicals would provide insights into disease mechanisms and new therapeutic approaches. We describe the first high-sensitivity, specific fluorescence probe for lipid radicals, 2,2,6-trimethyl-4-(4-nitrobenzo[1,2,5]oxadiazol-7-ylamino)-6-pentylpiperidine-1-oxyl (NBD-Pen). NBD-Pen directly detected lipid radicals in living cells by turn-on fluorescence. In a rat model of hepatic carcinoma induced by diethylnitrosamine (DEN), NBD-Pen detected lipid radical generation within 1 h of DEN administration. The lipid radical scavenging moiety of NBD-Pen decreased inflammation, apoptosis and oxidative stress markers at 24 h after DEN, and liver tumor development at 12 weeks. Thus, we have developed a novel fluorescence probe that provides imaging information about lipid radical generation and potential therapeutic benefits in vivo.

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Figure 1: Spectroscopic responses and selectivity for lipid radicals.
Figure 2: Liver cell fluorescence imaging and in vivo detection of lipid radicals produced during DEN metabolism.
Figure 3: OH-Pen, but not OH-Pen-NOMe (NOMe), suppresses DEN-induced HCC.
Figure 4: Influence of OH-Pen and OH-Pen-NOMe (NOMe) on hepatic injury and compensatory cell proliferation.
Figure 5

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Acknowledgements

This work was supported in part by JST PRESTO, by JSPS KAKENHI grant number 24390011 and 24659020, by the Platform Project for Supporting in Drug Discovery and Life Science Research from AMED, Japan, by the Suzuken Memorial Foundation, Nagoya, Japan, and by the Suntory Foundation for Life Sciences, Kyoto, Japan. We appreciate the technical support provided by the Research Support Center, Graduate School of Medical Sciences, Kyushu University.

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Author notes

  1. Toshihide Yamasaki

    Present address: Current address: Aix-Marseille Université, Institut de Chimie Radicalaire, Campus St.-Jérôme, Marseille, France.,

Authors and Affiliations

  1. Physical Chemistry for Life Science Laboratory, Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan

    Ken-ichi Yamada, Fumiya Mito, Yuta Matsuoka, Satsuki Ide, Kazushige Shikimachi, Ayano Fujiki, Daiki Kusakabe, Yuma Ishida, Masataka Enoki, Arisa Tada, Miyuki Ariyoshi & Toshihide Yamasaki

  2. Japan Science and Technology Agency, PRESTO, Saitama, Japan

    Ken-ichi Yamada

  3. Innovation Center for Medical Redox Navigation, Kyushu University, Fukuoka, Japan

    Mayumi Yamato

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Contributions

K.Y. designed experiments and conceived and supervised the overall project. K.Y. also performed final data analysis and wrote the manuscript. F.M., Y.M., K.S. and Y.I. performed experiments and analyzed data for properties and mechanistic analysis of probes. S.I., A.F. and A.T. performed experiments and analyzed data for animal experiments. D.K. and M.E. performed experiments and analyzed data for cell experiments. M.A. performed experiments early in the project. T.Y. designed and performed chemical syntheses and wrote the synthetic methods for the manuscript. M.Y. designed experiments and performed final data analysis.

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Correspondence to Ken-ichi Yamada.

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Supplementary Results, Supplementary Table 1 and Supplementary Figures 1–16. (PDF 19711 kb)

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Synthetic procedures (PDF 190 kb)

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Yamada, Ki., Mito, F., Matsuoka, Y. et al. Fluorescence probes to detect lipid-derived radicals. Nat Chem Biol 12, 608–613 (2016). https://doi.org/10.1038/nchembio.2105

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