Visualization of endogenous hypochlorous acid during ferroptosis based on Rhodamine B fluorescent probe

Area of Science:

• Biochemistry
• Cell Biology
• Medical Imaging

Background:

• Hypochlorous acid (HClO) is implicated in ferroptosis, but its precise role and molecular mechanisms remain unclear.
• Understanding HClO dynamics is crucial for elucidating ferroptosis pathways and developing targeted therapies.

Purpose of the Study:

• To engineer a sensitive fluorescence turn-on probe (RDM-577) for detecting intracellular HClO fluctuations during ferroptosis.
• To investigate the dynamic changes of HClO in ferroptosis and inflammatory conditions using the developed probe.

Main Methods:

• Development and characterization of RDM-577, a novel fluorescence turn-on probe for HClO detection.
• In vitro and in vivo validation of probe sensitivity, detection limit, and imaging capabilities.
• Application of the probe to monitor HClO levels in ferroptosis-induced cells and a mouse model of hepatitis.

Main Results:

• RDM-577 demonstrated significant fluorescence enhancement (F/F0 = 35) upon reaction with HClO, with a low detection limit (5.8 nM).
• The probe successfully tracked dynamic HClO changes in ferroptosis cells and identified inflammatory sites in vivo.
• Imaging revealed that glutathione (GSH) protects against GPX4 degradation induced by ferroptosis.

Conclusions:

• RDM-577 is an effective tool for sensitive and specific detection of HClO in biological systems, including live cells and animal models.
• The probe facilitates intravital imaging of HClO, offering new insights into ferroptosis molecular mechanisms.
• Findings suggest a protective role of GSH against ferroptosis-induced GPX4 degradation.