目的 评价丹参醇提物（SMEE）对维拉帕米诱导斑马鱼急性心力衰竭的保护作用，筛选活性单体并探讨其作用机制。方法 以2 dpf野生型AB斑马鱼和心脏荧光转基因斑马鱼Tg（cmlc2： EGFP）为研究对象，采用维拉帕米诱导急性心力衰竭，检测静脉窦-动脉球（SV-BA）距离、心包水肿、静脉淤血、心室短轴缩短率、每搏输出量、射血分数、心率、血流速度及nppa、nppb mRNA表达，评价SMEE（10、20、40 μg·mL^-1）的心脏保护作用。采用UPLC-Q-TOF/MS表征SMEE主要化学成分，结合网络药理学、Lipinski规则及结构相似性分析优选候选活性单体，并利用斑马鱼心力衰竭模型进行验证。采用H₂O₂诱导AC16心肌细胞损伤，给予迷迭香酸（10、20、40 μg·mL^-1）处理24 h，检测细胞活力、B型钠尿肽（BNP）、心房钠尿肽（ANP）含量及总超氧化物歧化酶（T-SOD）活性；结合蛋白质组学筛选相关通路及蛋白，采用Western blotting检测PDZ和LIM结构域蛋白2（PDLIM2）、核因子-κB p65（NF-κB p65）及p-NF-κB p65蛋白表达。结果 与对照组相比，模型组SV-BA距离、心包水肿面积、静脉瘀血面积及nppa、nppb mRNA相对表达量升高，心室短轴缩短率、每搏输出量、射血分数、心率和血流速度降低（P＜0.001）；与模型组相比，高浓度SMEE组上述指标均显著改善（P＜0.001）。UPLC-Q-TOF/MS共获得50个主要成分的质谱表征结果，其中相对峰面积占比约1%及以上的成分15个，综合筛选后确定迷迭香酸为优先进行实验验证的候选活性单体。与模型组相比，各浓度迷迭香酸组SV-BA距离、心包水肿面积、静脉瘀血面积及nppa、nppb mRNA相对表达量均显著降低（P＜0.001）。在H₂O₂诱导的AC16细胞损伤模型中，模型组细胞活力和T-SOD活性降低，BNP、ANP含量升高（P＜0.001）；与模型组相比，迷迭香酸组上述指标均显著改善（P＜0.01、0.001）。蛋白质组学结果显示心肌细胞骨架相关通路富集，并筛选出PDLIM2作为后续验证蛋白； Western blotting结果显示，模型组PDLIM2蛋白表达升高，NF-κB p65和p-NF-κB p65蛋白表达降低（P＜0.01、0.001）；与模型组相比，迷迭香酸处理后NF-κB p65表达升高、PDLIM2表达降低（P＜0.001），p-NF-κB p65表达升高（P＜0.05）。结论 迷迭香酸可在体内外模型中发挥心脏保护作用，其机制可能与调节PDLIM2/NF-κB p65信号、减轻氧化应激性心肌细胞损伤有关。

Objective To evaluate the protective effect of the ethanol extract of Salvia miltiorrhiza (SMEE) against verapamil-induced acute heart failure in zebrafish, screen candidate active constituents, and investigate the underlying mechanism. Methods Wild-type AB zebrafish and cardiac fluorescent transgenic zebrafish Tg (cmlc2:EGFP) at 2 days post-fertilization (dpf) were used. Acute heart failure was induced by verapamil, and sinus venosus-bulbus arteriosus (SV-BA) distance, pericardial edema, venous congestion, ventricular fractional shortening, stroke volume, ejection fraction, heart rate, blood flow velocity, and nppa and nppb mRNA expression were measured to evaluate the cardioprotective effect of the SMEE (10, 20, and 40 μg·mL^-1). UPLC-Q-TOF/MS was used to characterize the major chemical constituents of the extract. Candidate active constituents were prioritized by integrating network pharmacology, Lipinski’s rule, and structural similarity analysis, followed by validation in the zebrafish heart failure model. H₂O₂- induced injury was established in AC16 cardiomyocytes, after being treated with rosmarinic acid (10, 20, 40 μg·mL^-1) for 24 h, and cell viability, B-type natriuretic peptide (BNP) and atrial natriuretic peptide (ANP) contents, and total superoxide dismutase (T-SOD) activity were determined. Proteomics was performed to screen relevant pathways and proteins, and Western blotting was used to determine the protein expression of PDZ and LIM domain protein 2 (PDLIM2), nuclear factor-κB p65 (NF-κB p65), and phosphorylated NF-κB p65 (p-NF-κB p65). Results Compared with the control group, the model group showed increased SV-BA distance, pericardial edema area, venous congestion area, and nppa and nppb mRNA expression, together with decreased ventricular fractional shortening, stroke volume, ejection fraction, heart rate, and blood flow velocity (P < 0.001). Compared with the model group, all of the above indices were significantly improved in the high-concentration SMEE group (P < 0.001). A total of 52 major constituents were characterized by UPLC-Q-TOF/MS, of which 15 had relative peak areas of approximately 1% or higher. Integrated screening prioritized rosmarinic acid as the candidate active constituent for experimental validation. Compared with the model group, all concentrations of rosmarinic acid significantly reduced SV-BA distance, pericardial edema area, venous congestion area, and nppa and nppb mRNA expression in zebrafish (P < 0.001). In the H₂O₂-induced AC16 cell injury model, the model group exhibited decreased cell viability and T-SOD activity and increased BNP and ANP contents (P < 0.001). Compared with the model group, all of these indices were significantly improved by the medium-concentration rosmarinic acid treatment (P < 0.001) and by the high-concentration treatment (P < 0.01). Proteomic analysis showed enrichment of cardiomyocyte cytoskeleton-related pathways and identified PDLIM2 as a protein for subsequent validation. WB analysis showed that PDLIM2 protein expression was increased, whereas NF-κB p65 and p-NF-κB p65 protein expression was decreased in the model group. Compared with the model group, rosmarinic acid treatment increased NF-κB p65 expression and decreased PDLIM2 expression (P < 0.001), while increasing p-NF-κB p65 expression (P < 0.05). Conclusion Rosmarinic acid exerts cardioprotective effects in both in vivo and in vitro models, and its mechanism may be associated with regulation of PDLIM2/NF-κB p65 signaling and attenuation of oxidative stress-induced cardiomyocyte injury.

R965

国家重点研发计划“战略性科技创新合作”重点专项（2026YFE0201900）；国家自然基金面上基金（82574783）；泰山学者青年专家项目（tsqn202312233）；山东第一医科大学（山东省医学科学院）临床-基础联合创新团队项目（202404）