A Highly Sensitive
Ratiometric Fluorescent Probe for
the Detection of Cytoplasmic and Nuclear Hydrogen Peroxide
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Abstract
As a marker for oxidative stress
and a second messenger in signal
transduction, hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) plays
an important role in living systems. It is thus critical to monitor
the changes in H<sub>2</sub>O<sub>2</sub> in cells and tissues. Here,
we developed a highly sensitive and versatile ratiometric H<sub>2</sub>O<sub>2</sub> fluorescent probe (<b>NP1</b>) based on 1,8-naphthalimide
and boric acid ester. In response to H<sub>2</sub>O<sub>2</sub>, the
ratio of its fluorescent intensities at 555 and 403 nm changed 1020-fold
within 200 min. The detecting limit of <b>NP1</b> toward H<sub>2</sub>O<sub>2</sub> is estimated as 0.17 μM. It was capable
of imaging endogenous H<sub>2</sub>O<sub>2</sub> generated in live
RAW 264.7 macrophages as a cellular inflammation response, and especially,
it was able to detect H<sub>2</sub>O<sub>2</sub> produced as a signaling
molecule in A431 human epidermoid carcinoma cells through stimulation
by epidermal growth factor. This probe contains an azide group and
thus has the potential to be linked to various molecules via the click
reaction. After binding to a Nuclear Localization Signal peptide,
the peptide-based combination probe (<b>pep-NP1</b>) was successfully
targeted to nuclei and was capable of ratiometrically detecting nuclear
H<sub>2</sub>O<sub>2</sub> in living cells. These results indicated
that <b>NP1</b> was a highly sensitive ratiometric H<sub>2</sub>O<sub>2</sub> dye with promising biological applications