Method for screening DOX (doxorubicin)-cardiotoxicity-resistant active substances through two-color fluorescence labeling

Abstract

The invention discloses a method for screening DOX (doxorubicin)-cardiotoxicity-resistant active substances through two-color fluorescence labeling. The method comprises the following steps: myocardial cells are taken and added into to-be-screened substances for pre-protection; DOX is added for damage after the pre-protection is finished; a cell membrane integrity fluorescent probe and a cell nucleus fluorescent probe which have different excitation and emission wavelengths are added simultaneously into the damaged myocardial cells for fluorescence labeling; fluorescence images of the cells are collected and analyzed after dying is finished, and the protection rates of the to-be-screened substances for cell membranes and cell nucleuses of the myocardial cells are calculated; and the active substances whose protection rates for the cell membranes and cell nucleuses of the myocardial cells are all higher than 15% are screened out from the to-be-screened substances. Two fluorescent probes are adopted to label the myocardial cells simultaneously, the protection effects of the to-be-screened substances for the myocardial cells are surveyed from different action mechanisms, the screening and evaluation method has the characteristics of fastness, economy, high throughput and high content, and the screening and evaluation results are more comprehensive and precise.

Description

technical field [0001] The invention belongs to the field of drug screening and evaluation methods, and in particular relates to a method for screening anti-doxorubicin myocardial toxicity active substances with two-color fluorescent labels. Background technique [0002] Doxorubicin (DOX) is one of the broad-spectrum anti-tumor drugs commonly used in clinical practice, and is widely used in the treatment of solid tumors, leukemia, lymphoma, breast cancer and other tumors. However, doxorubicin will produce cumulative, dose-dependent myocardial toxicity, and further develop into irreversible myocardial damage, eventually leading to congestive heart failure, thus seriously inhibiting its clinical application. [0003] The mechanism of doxorubicin-induced cardiotoxicity has not been fully elucidated so far. It is mainly believed that doxorubicin-induced cardiotoxicity is the result of multiple factors, including free radical damage, apoptosis, mitochondrial damage, calcium ion o...

AI Technical Summary

Problems solved by technology

[0007] The disadvantage of this method is that when single-color fluorescent labeling is used in drug screening research, the information obtained is single, which is not conducive to a comprehensive evaluation of the effect of compounds on cells, and there is a phenomenon of missing screening.

[0008] At present, there are no relevant reports introducing a high-content screening method for simultaneously labeling cells with multiple fluorescent dyes, and determining the degree of cell damage and the protective effect of drugs through cell fluorescence imaging

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