DataSheet2_Targeting the PANoptosis signaling pathway for myocardial protection: therapeutic potential of Xian Ling Gu Bao capsule.pdf

Introduction: Myocardial infarction (MI), the most prevalent ischemic heart disease, constitutes a primary cause of global cardiovascular disease with incidence and mortality. The pathogenesis of MI is exceedingly intricate, with PANoptosis playing a pivotal role in its pathological process. Xian Ling Gu Bao capsule (XLGB) contains various active components, including flavonoids, terpenes, and phenylpropanoids, and exhibits a wide range of pharmacological activities. However, it remains unclear whether XLGB can protect the myocardium from damage after MI. This study aimed to investigate the impact of XLGB on isoprenaline (ISO)-induced MI in mice and its potential mechanisms.
Methods: This study assessed the protective effects of XLGB against ISO-induced MI through techniques such as echocardiography, HE staining, Masson staining, and enzyme-linked immunosorbent assay (ELISA). Furthermore, the potential mechanisms of XLGB's protective effects on MI were explored using bioinformatics, molecular docking, and molecular dynamics simulations. These mechanisms were further validated through immunofluorescence staining and Western blotting.
Results: The results demonstrated that various doses of XLGB exhibited a significant reduction in myocardial injury induced by myocardial infarction. Intriguingly, higher dosages of XLGB displayed superior therapeutic efficacy compared to the positive control metoprolol. This protective effect is primarily achieved through the inhibition of oxidative stress and the inflammatory processes. Furthermore, we have elucidated that XLGB protected the myocardium from MI-induced damage by suppressing PANoptosis, with a critical role played by the NLRP3/Caspase3/RIP1 signaling pathway. Of particular note, the primary compounds of XLGB were found to directly interact with NLRP3/Caspase3/RIP1, a discovery further validated through molecular docking and molecular dynamics simulations. This suggests that NLRP3/Caspase3/RIP1 may be a therapeutic target for XLGB-induced myocardial protection.
Conclusion: In summary, our findings reveal a novel property of XLGB: reverses myocardial damage following MI by inhibiting the NLRP3/Caspase3/RIP1-mediated PANoptosis pathway.

引言：心肌梗死（Myocardial infarction, MI）是最为常见的缺血性心脏病，是全球范围内心血管疾病发病与死亡的首要原因。MI的发病机制极为复杂，其中泛凋亡（PANoptosis）在其病理进程中发挥关键作用。仙灵骨葆胶囊（Xian Ling Gu Bao capsule, XLGB）含有黄酮类、萜类、苯丙素类等多种活性成分，具备广泛的药理活性。然而，XLGB是否能够保护心肌免受MI发生后的损伤仍不明确。本研究旨在探讨XLGB对异丙肾上腺素（isoprenaline, ISO）诱导的小鼠MI模型的保护作用及其潜在机制。

方法：本研究通过超声心动图、HE染色、Masson染色及酶联免疫吸附试验（enzyme-linked immunosorbent assay, ELISA）等技术，评估了XLGB对ISO诱导的小鼠MI的保护作用。此外，本研究借助生物信息学、分子对接及分子动力学模拟，探索了XLGB发挥心肌保护作用的潜在机制，并通过免疫荧光染色与蛋白质印迹（Western blotting）对上述机制进行了验证。

结果：研究结果显示，不同剂量的XLGB均可显著减轻MI诱导的心肌损伤。值得关注的是，高剂量XLGB的治疗效果优于阳性对照药美托洛尔（metoprolol）。该保护作用主要通过抑制氧化应激与炎症反应实现。此外，本研究阐明XLGB可通过抑制泛凋亡（PANoptosis）保护心肌免受MI诱导的损伤，其中NLRP3/Caspase3/RIP1信号通路发挥关键调控作用。尤为关键的是，本研究发现XLGB中的主要活性成分可直接与NLRP3/Caspase3/RIP1靶点相结合，这一发现通过分子对接与分子动力学模拟得到了进一步验证，提示NLRP3/Caspase3/RIP1或可作为XLGB发挥心肌保护作用的治疗靶点。

结论：综上，本研究揭示了XLGB的全新药理属性：其可通过抑制NLRP3/Caspase3/RIP1介导的泛凋亡（PANoptosis）通路，逆转MI发生后的心肌损伤。