Visualization of doxorubicin-induced oxidative stress in isolated cardiac myocytes

The paper presents high-resolution fluorescence images obtained using laser-scanning confocal microscopy. Isolated cells from adult rat hearts were preloaded with 2',7'-dichlorofluorescin (an oxidant-sensitive fluorescent probe) and exposed to doxorubicin, an important anticancer drug with prominent cardiotoxicity. Fluorescence images were collected from live cells simultaneously on two channels: 1) 515-530 nm emission range was used to monitor an increase in dichlorofluorescein, the oxidized product of dichlorofluorescin, and 2) emission > 610 nm was used to visualize the intracellular distribution of doxorubicin. The images reveal intracellular oxidation close to the mitochondria after only 20 min of exposure of isolated cardiomyocytes to 40-160 mu M doxorubicin. The data confirm an oxidative mechanism of doxorubicin cardiotoxicity and demonstrate the capability of a new technique to monitor intracellular oxidation in living cardiomyocytes.