Abstract 4139241: The Triple Medical Therapy Prevented Post-infarction Cardiac Rupture via Inhibiting MMPs Over-activation and Macrophage-induced Cardiac Fibroblasts Ferroptosis Through TRAF6/NF-κB/C/EBPβ Down-regulation mediated by different microRN

Background: Percutaneous coronary intervention has significantly improved the prognosis of STEMI, though there is still the risk of fetal mechanical complications, particularly cardiac rupture(CR), which remains an international clinical problem unresolved. Hypothesis: We hypothesized that the combined triple medical therapy( ANT ) with A torvastatin, N icorandil and Chinese patent medicine T ongxinluo(TXL) could be effective in post-infarction CR precautions via significantly inhibiting macrophage activation and secondary ferroptosis of cardiac fibroblasts(CFs) through TRAF6/NF-κB/C/EBPβ pathway. Methods: The 14-day survival and CR rates of AMI mice treated with Atorvastatin, Nicorandil and Tongxinluo, singly or in dual and triple combination, were all compared via the Kaplan-Meier curves. Then the inflammation level and infarct region were measured via ELISA, immunoblot and histopathology. Sequentially immunoprecipitation, luciferase report gene and transcriptome sequencing were introduced to understand the role of the TRAF6/NF-κB/C/EBPβ pathway and its upstream micro-RNAs in CR prevention. Further, in-vitro experiments were performed to demonstrate the crosstalk between macrophages activation and CFs ferroptosis in CR prevention. Results: Among all therapies involved, the triple combined ANT therapy supremely reduced the incidence of post-infarction CR (from 26.7% to 10.0%) and the mortality of AMI (from 30.0% to 13.3%), during which macrophages reduced most nuclear NF-κB p65 and C/EPBβ by nearly 70% simultaneously through miR215-5p-, miR122-5p-, miR-299b-3p-mediated TRAF6 inhibition, with 50%+ MMP9 cut and more extracellular matrix remaining, especially collagens, Syndecan-1, Laminin and Agrin. And, with the ANT administration, only 20% macrophages remained in peri-infarct areas, accompanied by the lowest serum inflammatory level. Furthermore, CF ferroptosis alleviated most, evidenced by the 4-fold GPX4 and 3-fold FSP-1 up-regulation. Two independent anti-ferroptotic system, GPX4 and FSP-1, worked equally in the nicorandil effect on CF survival, however, with GPX4 or FSP-1 overwhelmingly underlying atorvastatin or Tongxinluo protection against CF ferroptosis respectively. Conclusions: Our results indicated the ANT combination upmost alleviated the excessive excitation of M1 macrophages and its induced CF ferroptosis via prohibiting TRAF6-mediated NF-κB and C/EBPβ translocation, and facilitated myocardial repair to prevent post-infarction cardiac rupture onset.