A method for establishing the electrophysiological model of alcoholic cardiomyopathy.

OBJECTIVES

To establish an electrophysiological model of alcoholic cardiomyopathy by inducing pluripotent stem cells (iPSCs) to differentiate into cardiomyocytes (iPSC-CM) in vitro.

METHODS

The human iPSC were expanded in vitro and differentiated into iPSC-CM. The iPSC-CM were divided into a blank control group, an alcoholic experiment group (according to the concentration of alcoholic, the alcoholic experiment was also divided into many subgroups), and a KN93 treatment group. Then the efficiency of iPSC differentiated to iPSC-CM was detected by immunofluorescence, the function of iPSC-CM was detected by cell counting kit-8 (CCK8) assay and lactate dehydrogenase (LDH) activity assay kit. The electrophysiological activity of iPSC-CM was monitored by real time cellular analysis (RTCA), the injury of iPSC-CM caused by alcohol was further verified by the mitochondrial membrane potential fluorescence probe JC-1 staining combined with RTCA analysis.

RESULTS

Compared with the blank control group, the different doses (25, 50, 100, 150, 200, 250, 300 mmol/L) of alcohol could significantly inhibit the proliferation of iPSC-CM in a dose-dependent manner (all <0.05). Compared with the blank control group, the activity of iPSC-CM was significantly reduced by 100 mmol/L alcohol, resulting in the increase of LDH release, the decrease of mitochondrial membrane potential, the amplitude and beating rate (all <0.05). Compared with the 100 mg/mL alcoholic experiment group, the KN93 treatment group significantly alleviated the damage of alcohol to iPSC-CM by blocking the necrotic apoptotic pathway, resulting in the decrease of LDH release, the increase of mitochondrial membrane potential, the amplitude and beating rate (all <0.05).

CONCLUSIONS

The electrophysiological model of alcoholic cardiomyopathy based on the differentiation of cardiomyocytes are successfully established, which can be used to study the electrophysiological activity and the molecular mechanism for relevant diseases, and it may provide a more reasonable and effective research tool for drug screening and clinical study.