Ma Yi, Wu Yi, Xia Zhengchao, Li Jingyi, Li Xiaorong, Xu Pingxiang, Zhou Xuelin, Xue Ming
Department of Pharmacology, Beijing Laboratory for Biomedical Detection Technology and Instrument, School of Basic Medical Sciences, Capital Medical University, Beijing, China.
Beijing Tropical Medicine Research Institute, Beijing Friendship Hospital, Capital Medical University, Beijing, China.
Front Pharmacol. 2019 Nov 12;10:1356. doi: 10.3389/fphar.2019.01356. eCollection 2019.
The health supplement of () is well known for its effective properties against hypoxia. However, the mechanisms of its anti-hypoxic action were still unclear. The objective of this work was to evaluate the molecular mechanisms of RC extract against hypoxia in a hypoxic zebrafish model through metabonomics and network pharmacology analysis. The hypoxic zebrafish model in the environment with low concentration (3%) of oxygen was constructed and used to explore the anti-hypoxic effects of RC extract, followed by detecting the changes of the metabolome in the brain through liquid chromatography-high resolution mass spectrometry. An network for metabolite-protein interactions was further established to examine the potential mechanisms of RC extract, and the mRNA expression levels of the key nodes were validated by real-time quantitative PCR. As results, RC extract could keep zebrafish survive after 72-h hypoxia improving lactate dehydrogenase, citrate synthase, and hypoxia-induced factor-1α in brains. One hundred and forty-two differential metabolites were screened in the metabonomics, and sphingolipid metabolism pathway was significantly regulated after RC treatment. The constructed protein-metabolites network indicated that the HIF-related signals were recovered, and the mRNA level of AMPK was elevated. In conclusion, RC extract had markedly anti-hypoxic effects in zebrafish changing sphingolipid metabolism, HIF-related and AMPK signaling pathways.
（）的健康补充剂以其抗缺氧的有效特性而闻名。然而，其抗缺氧作用的机制仍不清楚。这项工作的目的是通过代谢组学和网络药理学分析，在缺氧斑马鱼模型中评估RC提取物抗缺氧的分子机制。构建了低氧浓度（3%）环境下的缺氧斑马鱼模型，用于探究RC提取物的抗缺氧作用，随后通过液相色谱-高分辨率质谱检测大脑中代谢组的变化。进一步建立了代谢物-蛋白质相互作用网络，以研究RC提取物的潜在机制，并通过实时定量PCR验证关键节点的mRNA表达水平。结果显示，RC提取物可使斑马鱼在缺氧72小时后存活，提高大脑中的乳酸脱氢酶、柠檬酸合酶和缺氧诱导因子-1α水平。代谢组学筛选出142种差异代谢物，RC处理后鞘脂代谢途径受到显著调节。构建的蛋白质-代谢物网络表明，HIF相关信号得以恢复，且AMPK的mRNA水平升高。总之，RC提取物在斑马鱼中具有显著的抗缺氧作用，改变了鞘脂代谢、HIF相关和AMPK信号通路。
