Ferulic acid in combination with ginsenoside Rb1 alleviates myocardial no-reflow by inhibiting platelet HMGB1 release and NET formation.

BACKGROUND: The no-reflow (NR) phenomenon remains a challenge in the treatment of acute myocardial infarction. This study aimed to explore the therapeutic potential and underlying mechanism of a combination of ferulic acid (FA) and ginsenoside Rb1 (Rb1), active components of the traditional Chinese herbal pair of Ligusticum chuanxiong Hort. and Panax ginseng C. A. Mey., respectively, in alleviating myocardial ischemia-reperfusion injury (MIRI) and NR. METHODS: A rat model of MIRI was established to evaluate the effects of FA and Rb1 on cardiac function, infarction/NR area, microthrombi formation, and serum biomarkers. An integrated strategy combining network pharmacology, molecular docking, and molecular dynamics simulations was employed to predict key pathways and targets. Platelet HMGB1 release and neutrophil extracellular trap (NET) formation were investigated both in vitro and vivo. RESULTS: MIRI induced obvious NR, accompanied by enhanced platelet HMGB1 release, increased NET formation and microthrombi accumulation. Bioinformatical analyses confirmed that FA and Rb1 stably interacts with HMGB1 and PAD4. Experimentally, FA predominantly inhibited platelet HMGB1 release, with IC(50) of 19.28 µM, by suppressing the p38/ERK1/2 pathway. Rb1 exhibited stronger efficacy in inhibiting PAD4 enzyme activity. The FA-Rb1 combination demonstrated superior effects compared to either agent alone, effectively suppressing NET formation, improving cardiac function, and reducing both NR area and microthrombi burden. CONCLUSIONS: The combination of FA and Rb1 not only inhibits platelet HMGB1 release but also reduces NETs, thereby enhancing anti-NR efficacy. These findings propose a novel therapeutic approach involving FA-Rb1 combination therapy for alleviating myocardial NR.