There is an urgent need to elucidate the pathogenesis of myocardial ischemia (MI) and potential drug treatments. Here, the anti-MI mechanism and material basis of Ginkgo biloba L. extract (GBE) were studied from the perspective of energy metabolism flux regulation. Metabolic flux analysis (MFA) was performed to investigate energy metabolism flux disorder and the regulatory nodes of GBE components in isoproterenol (ISO)-induced ischemia-like cardiomyocytes. It showed that [U–¹³C] glucose derived m+2 isotopologues from the upstream tricarboxylic acid (TCA) cycle metabolites were markedly accumulated in ISO-injured cardiomyocytes, but the opposite was seen for the downstream metabolites, while their total cellular concentrations were increased. This indicates a blockage of carbon flow from glycolysis and enhanced anaplerosis from other carbon sources. A Seahorse test was used to screen for GBE components with regulatory effects on mitochondrial aerobic respiratory dysfunction. It showed that bilobalide protected against impaired mitochondrial aerobic respiration. MFA also showed that bilobalide significantly modulated the TCA cycle flux, reduced abnormal metabolite accumulation, and balanced the demand of different carbon sources. Western blotting and PCR analysis showed that bilobalide decreased the enhanced expression of key metabolic enzymes in injured cells. Bilobalide's efficacy was verified by in vivo experiments in rats. This is the first report to show that bilobalide, the active ingredient of GBE, protects against MI by rescuing impaired TCA cycle flux. This provides a new mechanism and potential drug treatment for MI. It also shows the potential of MFA/Seahorse combination as a powerful strategy for pharmacological research on herbal medicine.

迫切需要阐明心肌缺血 (MI) 的发病机制和潜在的药物治疗。在此，从能量代谢通量调节的角度研究了银杏叶提取物（GBE）的抗心肌梗死机制和物质基础。进行代谢通量分析（MFA）以研究能量代谢通量障碍和异丙肾上腺素（ISO）诱导的缺血样心肌细胞中GBE成分的调节节点。它表明 [U- ¹³C] 来自上游三羧酸 (TCA) 循环代谢物的葡萄糖衍生的 m+2 同位素体在 ISO 损伤的心肌细胞中显着积累，但下游代谢物的情况相反，而它们的总细胞浓度增加。这表明来自糖酵解的碳流阻塞和来自其他碳源的增强的回补。Seahorse 测试用于筛选对线粒体有氧呼吸功能障碍具有调节作用的 GBE 成分。结果表明，bilobalide 可以防止线粒体有氧呼吸受损。MFA 还表明，bilobalide 显着调节 TCA 循环通量，减少异常代谢物积累，平衡不同碳源的需求。Western印迹和PCR分析表明，bilobalide降低了受损细胞中关键代谢酶的表达增强。Bilobalide 的功效已通过大鼠体内实验得到验证。这是第一份表明 GBE 的活性成分 bilobalide 通过挽救受损的 TCA 循环通量来预防 MI 的报告。这为 MI 提供了一种新的机制和潜在的药物治疗方法。它还显示了 MFA/Seahorse 组合作为草药药理学研究的有力策略的潜力。