Senkyunolide I and Danshensu in combination ameliorate post-MI myocardial fibrosis by regulating TGF-β1/PAI-1 in mice and heart organoids.

BACKGROUND

Myocardial fibrosis following myocardial infarction (MI) is a critical determinant of long-term clinical outcomes in heart disease. However, this central pathological process in the progression from MI to heart failure is not fully understood, posing challenges for targeted therapeutic options.

PURPOSE: This study aimed to identify new targets critical to myocardial fibrosis after MI and to evaluate the potential of Guanxinning injection (GXNI, a combination of Danshen and Chuanxiong) to suppress post-MI myocardial fibrosis.

METHODS

A MI mouse model was established by permanent ligation of the left anterior descending artery. Therapeutic effects were assessed by echocardiography and histopathology. HPLC chromatography, Network pharmacology, molecular docking, two-dimensional primary cell cultures, and iPS-induced cardiac organoid models were employed to identify active components and investigate underlying mechanisms.

RESULTS

GXNI reduced myocardial fibrosis and injury, improving cardiac function. Network pharmacology and molecular docking identified Senkyunolide I (SENI) and Danshensu (DSS) as the key active components acting via the TGF-β1/PAI-1 pathway. Thermal shift assay demonstrated direct binding of SENI and DSS to PAI-1. In vitro and cryoinjury cardiac organoid models confirmed that GXNI and the SENI-DSS combination inhibited fibroblast activation and TGF-β1/PAI-1 expression, reducing fibrosis.

CONCLUSION

We demonstrate for the first time that the TGF-β1/PAI-1 signaling pathway plays a critical role in post-MI myocardial fibrosis, and GXNI attenuates MI via this pathway. Secondly, the active components of GXNI, SENI and DSS, act combinatorially to enhance the anti-fibrotic therapeutic efficacy. Furthermore, cryoinjury cardiac organoids serve as a powerful platform for investigating the mechanisms underlying myocardial infarction.