Integrating Network Pharmacology and Transcriptome Sequencing Unravel the Therapeutic Mechanism of Zhilong Huoxue Tongyu Capsule for Treating Chronic Heart Failure

Introduction: Chronic heart failure represents a hallmark end-stage pathology of numerous cardiovascular diseases, posing a serious threat to human health and quality of life, often with a poor prognosis. Zhilong Huoxue Tongyu Capsule (ZLHXTY), a traditional Chinese medicine formula consisting of five herbs, has demonstrated clinical efficacy in improving cardiac function and reducing serum levels of NT-proBNP and hs-CRP in CHF patients. However, the underlying molecular mechanisms remain largely unclear.Methods: This study employed an integrated approach combining network pharmacology, RNA-Seq, and qRT-PCR. Network pharmacology analysis was performed using the BATMAN-TCM database. Parallel RNA-Seq was conducted on ZLHXTY-treated hiPS-CMs to identify differentially expressed genes (DEGs), the findings were validated by qRT-PCR. The aim was to systematically investigate the therapeutic mechanisms of ZLHXTY in CHF.Results: We identified 52 active components and 1063 potential targets of ZLHXTY. RNA-Seq revealed 7476 DEG in ZLHXTY-treated hiPS-CMs. An intersection analysis of these datasets revealed 184 key therapeutic targets, including AKT1, HSP90AA1, NCOR1, MED1, RXRB, PPARA, PRKCA, RARA, RXRG, and ESR1. Pathway enrichment analysis indicated that these targets are significantly involved in biological processes related to fibrosis inhibition, immune regulation, metabolic promotion, and cardiac functional improvement. Treatment with ZLHXTY significantly downregulated the expression of PPARA, PRKCA, and MYH7 in hiPS-CMs. The PI3K-Akt signaling pathway was identified as centrally involved in regulating these processes.Conclusions: Collectively, our findings demonstrate that ZLHXTY ameliorates CHF through multi-component, multi-target, and multi-pathway mechanisms, providing important support for its clinical application and further drug development.