Protective effect and mechanism of Qishen Yiqi Formula on mitochondrial structure and function of cardiomyocytes injured by H2O2 / 中草药

ABSTRACT

Objective: To establish a cell-based multi-parameter mitochondrial structure and functional evaluation system for hydrogen peroxide (H2O2) injury in cardiomyocytes and investigate the protective effect of Qishen Yiqi Formula on H9c2 cells and its mechanism. Methods: For the in vitro myocardial cell injury model, H9c2 cells were divided into control, model (H2O2), positive control (carbonyl cyanide-cyanomethoxy basic hydrazine, 5 μmol/L), and Qishen Yiqi Formula (0.2 mg/mL) groups, with three duplicates in each group, and were cultured in triplicate for 24 h with corresponding drugs, followed by 2 h H2O2 induction. H2O2 injury model was established with H9c2 cell line. Mitochondrial function and morphological texture were evaluated by Operetta high content imaging system. Mitochondrial respiration and bioenergetic metabolism states were measured by Seahorse Bioscience XF extracellular flux analyzer. Cardiomyocyte apoptosis were quantified by flow cytometry. Results: Qishen Yiqi Formula prevented the decrease of mitochondrial membrane potential and the increase of mitochondrial mass and improved mitochondrial morphological integrity. Functionally, Qishen Yiqi Formula increased basic oxygen consumption rate, ATP-linked oxygen consumption rate, maximum oxygen consumption, and the reserve capacity. It also reduced the apoptotic rate at early and end stage and increased the myocyte survival. Conclusion: Based on the structure and functional evaluation of the mitochondria, results indicated that the compound Qishen Yiqi Formula protected the H9c2 cells by improving the mitochondrial function and energy metabolism as well as reducing the apoptosis.