Fluorescence based Tool to Detect Endogenous Peroxynitrite in M1-Polarized Murine J774.2 Macrophages

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Abstract

Oxidative stress and inflammation are intrinsically linked to each other in addition they are implicated in the evolution and pro-gression of non-communicable diseases (NCD). Large amounts of reactive oxygen species (ROS) are generated as part of the im-mune response towards NCD. Among all the ROS species, peroxynitrite (ONOO-) has the shortest half-life with < 20 ms under typical physiological conditions. Hence, detecting ONOO- and studying its generation in vitro allows for a better understanding of inflammatory processes. We demonstrate that peroxyresorufin-1 (PR1) is a selective and sensitive ONOO- fluorescence based sen-sor in J774.2 macrophages. PR1 was able to detect changes in ONOO- production upon investigation of different factors: en-hanced generation of ONOO- through LPS and IFN- as well as diminished ONOO- production with the introduction of superox-ide and nitric oxide synthase scavengers. Our study validates PR1 as an effective tool for the detection of ONOO- in J774.2 mu-rine macrophages and should allow for further elucidation of ROS biology and chemistry.

Original languageEnglish
Pages (from-to)10621–10627
Number of pages7
JournalAnalytical Chemistry
Volume90
Issue number17
Early online date2 Aug 2018
DOIs
Publication statusPublished - 4 Sep 2018

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Peroxynitrous Acid
Macrophages
Reactive Oxygen Species
Fluorescence
Oxidative stress
Nitric Oxide Synthase

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

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title = "Fluorescence based Tool to Detect Endogenous Peroxynitrite in M1-Polarized Murine J774.2 Macrophages",
abstract = "Oxidative stress and inflammation are intrinsically linked to each other in addition they are implicated in the evolution and pro-gression of non-communicable diseases (NCD). Large amounts of reactive oxygen species (ROS) are generated as part of the im-mune response towards NCD. Among all the ROS species, peroxynitrite (ONOO-) has the shortest half-life with < 20 ms under typical physiological conditions. Hence, detecting ONOO- and studying its generation in vitro allows for a better understanding of inflammatory processes. We demonstrate that peroxyresorufin-1 (PR1) is a selective and sensitive ONOO- fluorescence based sen-sor in J774.2 macrophages. PR1 was able to detect changes in ONOO- production upon investigation of different factors: en-hanced generation of ONOO- through LPS and IFN- as well as diminished ONOO- production with the introduction of superox-ide and nitric oxide synthase scavengers. Our study validates PR1 as an effective tool for the detection of ONOO- in J774.2 mu-rine macrophages and should allow for further elucidation of ROS biology and chemistry.",
author = "Maria Weber and Mackenzie, {Amanda B.} and Bull, {Steven D.} and James, {Tony D.}",
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T1 - Fluorescence based Tool to Detect Endogenous Peroxynitrite in M1-Polarized Murine J774.2 Macrophages

AU - Weber, Maria

AU - Mackenzie, Amanda B.

AU - Bull, Steven D.

AU - James, Tony D.

PY - 2018/9/4

Y1 - 2018/9/4

N2 - Oxidative stress and inflammation are intrinsically linked to each other in addition they are implicated in the evolution and pro-gression of non-communicable diseases (NCD). Large amounts of reactive oxygen species (ROS) are generated as part of the im-mune response towards NCD. Among all the ROS species, peroxynitrite (ONOO-) has the shortest half-life with < 20 ms under typical physiological conditions. Hence, detecting ONOO- and studying its generation in vitro allows for a better understanding of inflammatory processes. We demonstrate that peroxyresorufin-1 (PR1) is a selective and sensitive ONOO- fluorescence based sen-sor in J774.2 macrophages. PR1 was able to detect changes in ONOO- production upon investigation of different factors: en-hanced generation of ONOO- through LPS and IFN- as well as diminished ONOO- production with the introduction of superox-ide and nitric oxide synthase scavengers. Our study validates PR1 as an effective tool for the detection of ONOO- in J774.2 mu-rine macrophages and should allow for further elucidation of ROS biology and chemistry.

AB - Oxidative stress and inflammation are intrinsically linked to each other in addition they are implicated in the evolution and pro-gression of non-communicable diseases (NCD). Large amounts of reactive oxygen species (ROS) are generated as part of the im-mune response towards NCD. Among all the ROS species, peroxynitrite (ONOO-) has the shortest half-life with < 20 ms under typical physiological conditions. Hence, detecting ONOO- and studying its generation in vitro allows for a better understanding of inflammatory processes. We demonstrate that peroxyresorufin-1 (PR1) is a selective and sensitive ONOO- fluorescence based sen-sor in J774.2 macrophages. PR1 was able to detect changes in ONOO- production upon investigation of different factors: en-hanced generation of ONOO- through LPS and IFN- as well as diminished ONOO- production with the introduction of superox-ide and nitric oxide synthase scavengers. Our study validates PR1 as an effective tool for the detection of ONOO- in J774.2 mu-rine macrophages and should allow for further elucidation of ROS biology and chemistry.

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