Elucidation of the Mechanisms and Molecular Targets of Qishen Yiqi Formula for the Treatment of Pulmonary Arterial Hypertension using a Bioinformatics/Network Topology-based Strategy.

BACKGROUND AND OBJECTIVE: Qishen Yiqi formula (QSYQ) is used to treat cardiovascular disease in the clinical practice of traditional Chinese medicine. However, few studies have explored whether QSYQ affects pulmonary arterial hypertension (PAH), and the mechanisms of action and molecular targets of QSYQ for the treatment of PAH are unclear. A bioinformatics/network topology-based strategy was used to identify the bioactive ingredients, putative targets, and molecular mechanisms of QSYQ in PAH. METHODS: A network pharmacology-based strategy was employed by integrating active component gathering, target prediction, PAH gene collection, network topology, and gene enrichment analysis to systematically explore the multicomponent synergistic mechanisms. RESULTS: In total, 107 bioactive ingredients of QSYQ and 228 ingredient targets were identified. Moreover, 234 PAH-related differentially expressed genes with a |fold change| >2 and an adjusted P value < 0.005 were identified between the PAH patient and control groups, and 266 therapeutic targets were identified. The pathway enrichment analysis indicated that 85 pathways, including the PI3K-Akt, MAPK, and HIF-1 signaling pathways, were significantly enriched. TP53 was the core target gene, and 7 other top genes (MAPK1, RELA, NFKB1, CDKN1A, AKT1, MYC, and MDM2) were the key genes in the gene-pathway network based on the effects of QSYQ on PAH. CONCLUSION: An integrative investigation based on network pharmacology may elucidate the multicomponent synergistic mechanisms of QSYQ in PAH and lay a foundation for further animal experiments, human clinical trials and rational clinical applications of QSYQ.

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

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.