Establishment of Myocardial Oxidative Stress Model

Oxidative stress is a condition that produces reactive oxygen species (ROS) or free radicals (O₂^-, H₂O₂, and -OH) outside or inside cells and has toxic effects on cells. The heart is one of the main organs affected by ROS. Recent evidence suggests that oxidative stress is a common feature of many aspects of cardiovascular disease. During myocardial oxidative stress, the production of ROS is enhanced and the defense mechanism of cardiomyocytes is changed. The source of ROS in cardiomyocytes may be mitochondrial electron transport chain, nitric oxide synthase (NOS), NADPH oxidase, xanthine oxidase, lipoxygenase/cyclooxygenase, and the automatic oxidation of various substances.

Fig. 1 Role of the ROS system in the heart in physiology and disease. (D’Oria et al., 2020)

The establishment of the cardiomyocyte oxidative stress model is helpful to observe the effects of antioxidants or other substances on various oxidative stress indexes and apoptosis of cardiomyocytes, to reveal the protective effect of antioxidants on cardiomyocytes, or the toxicity of external substances on cardiomyocytes. As an expert in the field of cellular stress, Creative Bioarray can now establish cardiomyocyte oxidative stress models by many different means.

Sodium Arsenite Induction

Arsenic has cardiotoxicity. It can stimulate NADPH oxidase on the membrane of vascular endothelial cells and vascular smooth muscle cells in Vivo, increasing intracellular ROS levels. We used sodium arsenite to induce the oxidative stress model of H9c2 cardiomyocytes. After the establishment of the model, we detected the cell viability by CCK-8 colorimetry and the level of intracellular reactive oxygen species by dichlorofluorescein probe to determine the modeling effect.

High Glucose Induction

Human cardiomyocytes were cultured in a high glucose environment to establish an oxidative stress model of cardiomyocytes. The model can be used to study the complications of diabetes.

H₂O₂ Induction

H9c2 cardiomyocytes were routinely cultured and treated with H₂O₂ to establish an oxidative damage model. The model is suitable for studying and evaluating the ability of antioxidants to reduce oxidative damage to cardiomyocytes. We usually use CCK-8 colorimetry to detect cell viability, embracing flow cytometry to detect the apoptosis rate of stressed cells, and laser confocal combined with flow cytometry to observe the production of intracellular reactive oxygen species and the changes of mitochondrial membrane potential.

Hypoxia / Reoxygenation Induction

We induced oxidative stress injury of H9c2 cardiomyocytes by hypoxia/reoxygenation. This is a model simulating the surgical treatment process of reperfusion therapy for ischemic heart disease. The establishment of this model is helpful to find effective drugs against oxidative stress injury caused by reperfusion, which is of great significance to improve the treatment of ischemic heart disease.

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Reference

D’Oria, R., Schipani, R., Leonardini, A., Natalicchio, A., Perrini, S., & Cignarelli, A. et al. (2020). The Role of Oxidative Stress in Cardiac Disease: From Physiological Response to Injury Factor. Oxidative Medicine And Cellular Longevity, 2020, 1-29.

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