乌苏里瓦韦及其再生植株的利尿作用研究

Study on the Diuretic Effect of Lepisorus Ussuriensis (Regel et Maack) Ching and Its Regenerated Plants

关键词:  网络药理学  分子对接  乌苏里瓦韦  再生植株  利尿作用

Key words:  network pharmacology  molecular docking  Lepisorus ussuriensis (Regel et Maack) Ching  plantlet regeneration  diuretic activity

摘要:

目的 探讨乌苏里瓦韦利尿作用的机制，对比乌苏里瓦韦及其再生植株的利尿作用。方法 运用网络药理学探讨乌苏里瓦韦的利尿作用机制；利用AutoDock Vina1.1.2软件进行活性成分和核心靶点的分子对接，采用Pymol2.5.0软件对最优结果进行可视化；通过体内试验测量小鼠口服药物后4 h内尿量，评价乌苏里瓦韦及其再生植株的利尿作用。结果 从乌苏里瓦韦中的41个候选化合物中筛选得到共1 791个药物靶点，将删除重复靶点后的327个药物靶点与从GeneCards数据库中以“Diuretic”为关键词检索得到的1 324个疾病靶点作Venn图，共得到119个交集基因。PPI网络节点度值排名前6的核心蛋白为MAPK3、VEGFA、AKT1、IL6、ALB、TNF，通过拓扑分析得到度值排名前3的候选化合物分子黄芩素、香叶木素、山柰酚。GO功能分析与KEGG通路富集分析结果表明，乌苏里瓦韦活性成分利尿作用机制主要与response to toxic substance、membrane raft等生物过程有关，参与的主要信号通路有神经活性配体-受体相互作用、HIF-1信号通路、氮代谢等。分子对接结果显示黄芩素、香叶木素及山柰酚均可与核心靶点进行较好的自发结合。体内试验中乌苏里瓦韦组（9.5 g·kg^-1）尿量增加了1.26倍，K⁺增加了1.31倍，Cl^-增加了1.24倍。乌苏里瓦韦再生植株组（9.5 g·kg^-1）尿量增加了1.26倍，K⁺增加了1.29倍，Cl^-增加了1.26倍。结论 网络药理学结果表明，乌苏里瓦韦利尿作用可能是多组分、多靶点、多途径作用的结果；通过体内药理试验验证了乌苏里瓦韦及乌苏里瓦韦再生植株具有相似的利尿作用。

Abstract:

OBJECTIVE To explore the mechanism of diuretic effect of Lepisorus ussuriensis (Regel et Maack) Ching and to compare the diuretic effect of Lepisorus ussuriensis (Regel et Maack) Ching and its regenerated plants. METHODS Using network pharmacology to explore the diuretic mechanism of Lepisorus ussuriensis (Regel et Maack) Ching; AutoDock Vina1.1.2 was applied to acquire the molecular docking result of active components and core targets, then Pymol2.5.0 software was used to visualize the optimal results; the urine volume of mice was measured within 4 h after oral administration by in vivo experiments; to evaluate the diuretic effects of the Lepisorus ussuriensis (Regel et Maack) Ching and its regenerated plants. RESULTS A total of 1 791 drug targets were obtained from 41 candidate compounds of Lepisorus ussuriensis (Regel et Maack) Ching, and to map the 327 drug targets obtained after deleting repeated targets and the 1 324 disease targets retrieved from the GeneCards database with "Diuretic" as the key word. A total of 119 intersecting genes were obtained. The top 6 core proteins of PPI network nodes were MAPK3, VEGFA, AKT 1, IL6, ALB and TNF. The top three candidate compounds were baicalein(44), diosmetin(43) and kaempferol(43). The results of GO function analysis and KEGG pathway enrichment analysis showed that the diuretic effect of Lepisorus ussuriensis (Regel et Maack) Ching was mainly related to response to toxic substance, membrane raft et al, the main signal pathways involved were neuroactive ligand-receptor interaction, HIF-1 signaling pathway, nitrogen metabolism and so on. The results of molecular docking revealed that baicalein, diosmetinand, kaempferol had better spontaneous binding effect with the key target. The result of in vivo experiments showed that Lepisorus ussuriensis (Regel et Maack) Ching(9.5 g·kg^-1) increased the urinary volume(1.26-fold), as well as the excretion of K⁺(1.31-fold) and Cl^-(1.24-fold). The regenerated plants(9.5 g·kg^-1) increased the urinary volume(1.26-fold), as well as the excretion of K⁺(1.29-fold) and Cl^-(1.26-fold). CONCLUSION The mechanism of diuretic action of Lepisorus ussuriensis (Regel et Maack) Ching is studied by network pharmacology, and the characteristics of multi-component, multi-target and multi-pathway were verified. Lepisorus ussuriensis (Regel et Maack) Ching and its regenerated plants show the same diuretic activity through in vivo experiments.