DataSheet3_Astragaloside IV promotes pharmacological effect of Descurainia sophia seeds on isoproterenol-induced cardiomyopathy in rats by synergistically modulating the myosin motor.XLSX

Descurainia sophia seeds (DS), Astragalus mongholicus (AM), and their formulas are widely used to treat heart failure caused by various cardiac diseases in traditional Chinese medicine practice. However, the molecular mechanism of action of DS and AM has not been completely understood. Herein, we first used mass spectrometry coupled to UPLC to characterize the chemical components of DS and AM decoctions, then applied MS-based quantitative proteomic analysis to profile protein expression in the heart of rats with isoproterenol-induced cardiomyopathy (ISO-iCM) before and after treated with DS alone or combined with AM, astragaloside IV (AS4), calycosin-7-glucoside (C7G), and Astragalus polysaccharides (APS) from AM. We demonstrated for the first time that DS decoction alone could reverse the most of differentially expressed proteins in the heart of the rats with ISO-iCM, including the commonly recognized biomarkers natriuretic peptides (NPPA) of cardiomyopathy and sarcomeric myosin light chain 4 (MYL4), relieving ISO-iCM in rats, but AM did not pronouncedly improve the pharmacological efficiency of DS. Significantly, we revealed that AS4 remarkably promoted the pharmacological potency of DS by complementarily reversing myosin motor MYH6/7, and further downregulating NPPA and MYL4. In contrast, APS reduced the efficiency of DS due to upregulating NPPA and MYL4. These findings not only provide novel insights to better understanding in the combination principle of traditional Chinese medicine but also highlight the power of mass spectrometric proteomics strategy combined with conventional pathological approaches for the traditional medicine research.

播娘蒿籽（Descurainia sophia seeds, DS）与蒙古黄芪（Astragalus mongholicus, AM）及其复方制剂在中医临床中被广泛用于治疗各类心脏疾病引发的心力衰竭。然而，播娘蒿籽与蒙古黄芪的分子作用机制尚未完全阐明。本研究首先采用超高效液相色谱-质谱联用技术（UPLC-MS）对播娘蒿汤剂与蒙古黄芪汤剂的化学成分进行表征，随后采用基于质谱的定量蛋白质组学分析，对异丙肾上腺素诱导型心肌病（ISO-iCM）模型大鼠在单独使用播娘蒿汤剂，或联合使用蒙古黄芪、黄芪甲苷IV（astragaloside IV, AS4）、毛蕊异黄酮-7-葡萄糖苷（calycosin-7-glucoside, C7G）以及蒙古黄芪来源的黄芪多糖（Astragalus polysaccharides, APS）干预前后的心脏蛋白质表达谱进行了分析。本研究首次证实，单独使用播娘蒿汤剂即可逆转异丙肾上腺素诱导型心肌病模型大鼠心脏中绝大多数差异表达蛋白，包括心肌病公认的生物标志物利钠肽（NPPA）与肌节肌球蛋白轻链4（MYL4），从而缓解大鼠ISO-iCM症状，但蒙古黄芪并未显著提升播娘蒿汤剂的药理效能。尤为重要的是，本研究揭示黄芪甲苷IV可通过互补逆转肌球蛋白重链MYH6/7的表达，并进一步下调NPPA与MYL4的水平，从而显著增强播娘蒿汤剂的药理效能。与之相反，黄芪多糖可通过上调NPPA与MYL4的表达，降低播娘蒿汤剂的药理效能。本研究结果不仅为阐明中医复方配伍原则提供了全新视角，同时也凸显了质谱蛋白质组学策略联合常规病理学方法在中药研究中的应用价值。