Abstract：
BACKGROUND: Dengzhan Shengmai capsule (DZSM) is a traditional herb medicine used by Dai, an ethnic-minority community living in Xishuang banna tropical rainforest in Southwest of China. It was originally intended to treat disorders caused by insufficient brain function, characterized by gibberish, unresponsiveness, or confusion. Accumulating clinical evidences exhibited that it is effective on treating ischemic stroke (IS). However, the action of DZSM against IS needs to be further elucidated.
OBJECTIVE: To investigate the effect of DZSM and its active components against IS and the way of its action by multi-omics and network pharmacology.
METHODS: A middle cerebral artery occlusion/reperfusion (MCAO/R) rat model was established to investigate the effect of DZSM on the focal cerebral ischemia/reperfusion injury. An integrated strategy combining metabolomics, network pharmacology and transcriptomics was performed to systematically clarify the underlying mechanism of action of DZSM against IS. AutoDock Vina was applied to conduct molecular docking simulation for the binding between the potential active compounds and targets. Arachidonic acid (AA) induced platelet aggregation and lipopolysaccharide (LPS) stimulated microglial cells BV2 inflammation models were applied for the in vitro validation of effects of DZSM and its potential active compounds.
RESULTS: In MCAO/R rats, DZSM could significantly reduce the infarct volume. Putative target prediction and functional enrichment analysis based on network pharmacological indicated that the key targets and the potential active compounds played important roles in DZSM\'s treatment to IS. The targets included four common genes (PTGS1, PTGS2, NFKB1 and NR1I2) and five key TFs (NFKB1, RELA, HIF1A, ESR1 and HDAC1), whilst 22 potential active compounds were identified. Molecular docking indicated that good binding affinity have been seen between those compounds and NR1I2, NFKB1, and RELA. Multi-omics study revealed that DZSM could regulate glutamate by influencing citrate cycle and glutamate involved pathways, and have showed neuroprotection activity and anti-inflammation activity by inhibiting NF-κB pathway. Neuroprotective effects of DZSM was validated by regulating of NF-κB signaling pathway and its downstream NO, TNF-α and IL-6 cytokines contributed to the activity of DZSM and its active compounds of scutellarin, quercetin 3-O-glucuronide, ginsenoside Rb1, schizandrol A and 3, 5-diCQA, whilst the antithrombotic activity of DZSM and its active compounds of schisanhenol, apigenin and schisantherin B were screened out by anti-platelet aggregation experiment.
CONCLUSIONS: DZSM could against IS via regulating its downstream NO, TNF-α and IL-6 cytokines through NF-κB signaling pathway and alleviating thrombosis.
OBJECTIVE: To investigate the effect of DZSM and its active components against IS and the way of its action by multi-omics and network pharmacology.
METHODS: A middle cerebral artery occlusion/reperfusion (MCAO/R) rat model was established to investigate the effect of DZSM on the focal cerebral ischemia/reperfusion injury. An integrated strategy combining metabolomics, network pharmacology and transcriptomics was performed to systematically clarify the underlying mechanism of action of DZSM against IS. AutoDock Vina was applied to conduct molecular docking simulation for the binding between the potential active compounds and targets. Arachidonic acid (AA) induced platelet aggregation and lipopolysaccharide (LPS) stimulated microglial cells BV2 inflammation models were applied for the in vitro validation of effects of DZSM and its potential active compounds.
RESULTS: In MCAO/R rats, DZSM could significantly reduce the infarct volume. Putative target prediction and functional enrichment analysis based on network pharmacological indicated that the key targets and the potential active compounds played important roles in DZSM\'s treatment to IS. The targets included four common genes (PTGS1, PTGS2, NFKB1 and NR1I2) and five key TFs (NFKB1, RELA, HIF1A, ESR1 and HDAC1), whilst 22 potential active compounds were identified. Molecular docking indicated that good binding affinity have been seen between those compounds and NR1I2, NFKB1, and RELA. Multi-omics study revealed that DZSM could regulate glutamate by influencing citrate cycle and glutamate involved pathways, and have showed neuroprotection activity and anti-inflammation activity by inhibiting NF-κB pathway. Neuroprotective effects of DZSM was validated by regulating of NF-κB signaling pathway and its downstream NO, TNF-α and IL-6 cytokines contributed to the activity of DZSM and its active compounds of scutellarin, quercetin 3-O-glucuronide, ginsenoside Rb1, schizandrol A and 3, 5-diCQA, whilst the antithrombotic activity of DZSM and its active compounds of schisanhenol, apigenin and schisantherin B were screened out by anti-platelet aggregation experiment.
CONCLUSIONS: DZSM could against IS via regulating its downstream NO, TNF-α and IL-6 cytokines through NF-κB signaling pathway and alleviating thrombosis.
摘要：
背景：灯盏生脉胶囊（DZSM）是Dai使用的传统草药，生活在中国西南部西双版纳热带雨林的少数民族社区。它最初旨在治疗由脑功能不足引起的疾病，以胡言乱语为特征，反应迟钝,或混乱。积累的临床证据表明，它对治疗缺血性中风（IS）有效。然而,DZSM对IS的作用需要进一步阐明。
目的：通过多组学和网络药理学研究DZSM及其活性成分对IS的作用及其作用方式。
方法：建立大脑中动脉阻塞/再灌注（MCAO/R）大鼠模型，研究DZSM对局灶性脑缺血/再灌注损伤的影响。结合代谢组学的综合策略，进行了网络药理学和转录组学,以系统地阐明DZSM抗IS的潜在作用机制.利用AutoDockVina对潜在活性化合物与靶标之间的结合进行分子对接模拟。花生四烯酸（AA）诱导的血小板聚集和脂多糖（LPS）刺激的小胶质细胞BV2炎症模型用于体外验证DZSM及其潜在活性化合物的作用。
结果：在MCAO/R大鼠中，DZSM可显著减少梗死体积。基于网络药理学的推定靶点预测和功能富集分析表明,关键靶点和潜在活性化合物在DZSM治疗IS中起着重要作用。目标包括四个常见基因（PTGS1，PTGS2，NFKB1和NR1I2）和五个关键TF（NFKB1，RELA，HIF1A,ESR1和HDAC1），同时鉴定了22种潜在的活性化合物。分子对接表明在那些化合物与NR1I2、NFKB1和RELA之间已经看到良好的结合亲和力。多组学研究表明，DZSM可以通过影响柠檬酸盐周期和谷氨酸参与通路来调节谷氨酸，并通过抑制NF-κB通路表现出神经保护活性和抗炎活性。通过调节NF-κB信号通路及其下游NO,验证了DZSM的神经保护作用。TNF-α和IL-6细胞因子有助于DZSM及其灯盏乙素活性化合物的活性，槲皮素3-O-葡糖苷酸，人参皂苷Rb1，五味子醇A和3，5-diCQA，而DZSM及其五酚活性化合物的抗血栓形成活性，通过抗血小板聚集实验筛选出芹菜素和五味子素B。
结论：DZSM可以通过调节其下游NO来对抗IS，TNF-α和IL-6细胞因子通过NF-κB信号通路缓解血栓形成。
目的：通过多组学和网络药理学研究DZSM及其活性成分对IS的作用及其作用方式。
方法：建立大脑中动脉阻塞/再灌注（MCAO/R）大鼠模型，研究DZSM对局灶性脑缺血/再灌注损伤的影响。结合代谢组学的综合策略，进行了网络药理学和转录组学,以系统地阐明DZSM抗IS的潜在作用机制.利用AutoDockVina对潜在活性化合物与靶标之间的结合进行分子对接模拟。花生四烯酸（AA）诱导的血小板聚集和脂多糖（LPS）刺激的小胶质细胞BV2炎症模型用于体外验证DZSM及其潜在活性化合物的作用。
结果：在MCAO/R大鼠中，DZSM可显著减少梗死体积。基于网络药理学的推定靶点预测和功能富集分析表明,关键靶点和潜在活性化合物在DZSM治疗IS中起着重要作用。目标包括四个常见基因（PTGS1，PTGS2，NFKB1和NR1I2）和五个关键TF（NFKB1，RELA，HIF1A,ESR1和HDAC1），同时鉴定了22种潜在的活性化合物。分子对接表明在那些化合物与NR1I2、NFKB1和RELA之间已经看到良好的结合亲和力。多组学研究表明，DZSM可以通过影响柠檬酸盐周期和谷氨酸参与通路来调节谷氨酸，并通过抑制NF-κB通路表现出神经保护活性和抗炎活性。通过调节NF-κB信号通路及其下游NO,验证了DZSM的神经保护作用。TNF-α和IL-6细胞因子有助于DZSM及其灯盏乙素活性化合物的活性，槲皮素3-O-葡糖苷酸，人参皂苷Rb1，五味子醇A和3，5-diCQA，而DZSM及其五酚活性化合物的抗血栓形成活性，通过抗血小板聚集实验筛选出芹菜素和五味子素B。
结论：DZSM可以通过调节其下游NO来对抗IS，TNF-α和IL-6细胞因子通过NF-κB信号通路缓解血栓形成。
