Abstract：
This study aims to explore the anti-inflammatory, vasodilation, and cardioprotective effects of the intestinal absorption liquids containing Xinshubao Tablets or single herbs, and to elucidate the potential mechanism based on network pharmacology. Western blot was then conducted to validate the expression changes of core proteins. Lipopolysaccharide(LPS)-stimulated RAW264.7 cells were used to observe the anti-inflammatory effect. The vasodilation activity was examined by the microvessel relaxation assay in vitro. Oxygen-glucose deprivation(OGD)-induced H9c2 cells were used to investigate the cardioprotective effect. The chemical components were retrieved from Herb databases and composition of Xinshubao Tablets drug-containing intestinal absorption solution. Drug targets were retrieved from SwissTargetPrediction databases. GeneCards was searched for the targets associated with the anti-inflammatory, vasodilation, and cardioprotective effects. The common targets shared by the drug and the effects were used to establish the protein-protein interaction(PPI) network, from which the core targets were obtained. Finally, the core targets were imported into Cytoscape 3.9.1 for Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) analyses. The anti-inflammatory experiment showed that both Xinshubao Tablets and the single herbs constituting this formula had anti-inflammatory effects. Curcumae Radix had the strongest inhibitory effect on the production of tumor necrosis factor-α(TNF-α), and Salviae Miltiorrhizae Radix et Rhizoma had the strongest inhibitory effect on the generation of interleukin-6(IL-6). Xinshubao Tablets, Curcumae Radix, and Crataegi Fructus had vasodilation effect, and Crataegi Fructus had the strongest effect. Xinshubao Tablets, Salviae Miltiorrhizae Radix et Rhizoma, Acanthopanacis Senticosi Radix et Rhizoma seu Caulis, and Paeoniae Radix Alba had cardioprotective effects, and Salviae Miltiorrhizae Radix et Rhizoma had the strongest cardioprotective effect. Network pharmacology results demonstrated that except the whole formula, Salviae Miltiorrhizae Radix et Rhizoma had the most components with anti-inflammatory effect, and Curcumae Radix had the most components with vasodilation and cardioprotective effects, followed by Salviae Miltiorrhizae Radix et Rhizoma. The nitric oxide synthase 3(NOS3) was predicted as the core target for the anti-inflammatory, vasodilation, and cardioprotective effects. Western blot results showed that Xinshubao Tablets significantly up-regulated the expression of NOS3 in OGD-induced H9c2 cells. GO enrichment analysis showed that the effects were mainly related to lipid exported from cell, regulation of blood pressure, and inflammatory response. KEGG pathway enrichment predicted AGE-RAGE and HIF-1 signaling pathways as the key pathways.
摘要：
本研究旨在探索抗炎,血管舒张,和心脏保护作用的肠道吸收液含心舒宝片或单一草药，并基于网络药理学阐明潜在的机制。然后进行Western印迹以验证核心蛋白的表达变化。用脂多糖(LPS)刺激的RAW264.7细胞观察其抗炎作用。通过体外微血管松弛测定法检查血管舒张活性。氧糖剥夺（OGD）诱导的H9c2细胞用于研究心脏保护作用。从Herb数据库中检索化学成分和心舒堡片含药肠道吸收液的成分。从SwissTargetPrediction数据库检索药物靶标。寻找GeneCards与抗炎相关的目标，血管舒张,和心脏保护作用。药物共有的共同靶标和作用被用来建立蛋白质-蛋白质相互作用（PPI）网络，从中获得核心目标。最后,将核心靶标导入Cytoscape3.9.1进行基因本体论(GO)和京都基因和基因组百科全书(KEGG)分析.抗炎实验表明，心舒宝片和构成该配方的单味中药均具有抗炎作用。姜黄对肿瘤坏死因子-α(TNF-α)的产生抑制作用最强,丹参对白细胞介素-6(IL-6)的生成抑制作用最强。心舒宝片,姜黄,山楂有血管舒张作用，山楂的药效最强。心舒宝片,丹参，刺五加，白芍有心脏保护作用，丹参对心脏的保护作用最强。网络药理学结果表明，除了整个配方，丹参具有抗炎作用的成分最多,郁金香成分最具有血管舒张和心脏保护作用，其次是丹参。一氧化氮合酶3（NOS3）被预测为抗炎的核心靶标，血管舒张,和心脏保护作用。Westernblot结果显示，心舒宝片显著上调OGD诱导的H9c2细胞中NOS3的表达。GO富集分析表明,其作用主要与细胞外输的脂类有关,调节血压,和炎症反应。KEGG通路富集预测AGE-RAGE和HIF-1信号通路为关键通路。
