Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, PR China.
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, PR China.
Phytomedicine. 2018 Oct 1;49:83-94. doi: 10.1016/j.phymed.2018.06.026. Epub 2018 Jun 22.
Xin-Ke-Shu (XKS), a patent medicine consisting of five commonly used traditional Chinese herbs, is used for the treatment of coronary heart diseases. A previous study showed that XKS has protective effects for ameliorating myocardial ischemia/reperfusion (I/R) injury.
This study was aimed to deeply understand the mechanisms and compatible principle of XKS against hypoxia/reoxygenation (H/R) injury and the contribution of each single herb to the efficacy of XKS.
An H/R model in H9c2 cardiomyocytes was applied to mimic I/R injury observed in vivo. The cell viability, the levels of LDH, MDA, SOD, and apoptosis were determined to evaluate the cardioprotection of XKS and its subtracted formula (knocked out one herb) in H/R injury. Cell metabolomics, combined with western blot analysis, was performed to uncover the inert molecular mechanism of XKS against H/R injury.
Significant protective effects of XKS against oxidative stress and apoptosis induced by H/R injury were found in the pharmacodynamic evaluation. Moreover, the metabolic profile deviation of the H/R group from the control group was mainly ascribed to thirteen metabolites involved in four aberrant pathways, in which sphingolipid metabolism was revealed as the most relevant pathway involved in H/R injury (impact > 0.1). Notably, the accumulation of phytosphingosine (VIP = 5.84) was considered the most likely characteristic in H/R injury, which is well known to promote the opening of the mitochondrial permeability transition pore (mPTP) and activate cell apoptosis. Furthermore, XKS ameliorated all the abnormalities of the metabolic network in response to H/R injury. In agreement with this, a western blot analysis showed that XKS markedly regulated the over-expression of CaMK II and cleaved caspase-3. However, the subtracted formula showed no significant difference in comparison with the XKS group on protecting H/R injury except for QDS (subtracted Dan-Shen from XKS).
The roots of Salvia miltiorrhiza Bge. (Dan-Shen) play an important role in the regulation of Ca overloading, oxidative stress and apoptosis in H/R injury. Our study enabled information from holistic cell metabolomics to be used for mechanism and compatibility rule elucidations of TCMs.
心可舒（XKS）是一种由五种常用中药组成的专利药物，用于治疗冠心病。先前的研究表明，XKS 具有改善心肌缺血/再灌注（I/R）损伤的保护作用。
本研究旨在深入了解 XKS 抗缺氧/复氧（H/R）损伤的机制和配伍原则，以及每种单味中药对 XKS 疗效的贡献。
应用 H9c2 心肌细胞 H/R 模型模拟体内 I/R 损伤。测定细胞活力、LDH、MDA、SOD 水平和细胞凋亡，评价 XKS 及其减去配方（敲除一种草药）对 H/R 损伤的心脏保护作用。结合 Western blot 分析，进行细胞代谢组学研究，揭示 XKS 抗 H/R 损伤的潜在分子机制。
药效学评价发现 XKS 对 H/R 损伤诱导的氧化应激和细胞凋亡有显著的保护作用。此外，与对照组相比，H/R 组的代谢谱偏差主要归因于 13 种代谢物，涉及四个异常途径，其中鞘脂代谢被认为是与 H/R 损伤最相关的途径（影响>0.1）。值得注意的是，植物神经鞘氨醇（VIP=5.84）的积累被认为是 H/R 损伤最可能的特征，它被认为可促进线粒体通透性转换孔（mPTP）的开放，并激活细胞凋亡。此外，XKS 改善了代谢网络对 H/R 损伤的所有异常。Western blot 分析也显示，XKS 显著调节 CaMK II 的过度表达和 cleaved caspase-3。然而，与 XKS 组相比，减去配方（减去 XKS 中的丹参）除了 QDS（丹参）之外，在保护 H/R 损伤方面没有显著差异。
丹参的根在调节 H/R 损伤中的 Ca 超载、氧化应激和细胞凋亡方面发挥着重要作用。我们的研究使整体细胞代谢组学的信息能够用于中药的机制和配伍规律的阐明。
