目的 阐明藏药长毛风毛菊的药效及化学成分，探讨其有效成分、关键靶点和相关通路，明确其治疗肾性水肿的作用机制。方法 将SD大鼠分为空白组、阿霉素诱导肾性水肿模型组，长毛风毛菊给药低剂量组、高剂量组。测定血清生化指标，探讨各组总蛋白(total protein，TP)、尿素氮(blood u-rea nitrogen，BUN)、血肌酐(serum creatinine，SCr)、甘油三酯(triglyceride，TG)、总胆固醇(serum total cholestero，TC)、低密度脂蛋白(low density lipoprotein，LDL-C)、高密度脂蛋白(high density lipopro-tein，HDL-C)指标的情况。采用UPLC-Q-TOF-MS/MS技术对藏药长毛风毛菊的化学成分进行分析;在SwissTargetPrediction、GeneCards数据库分别获取长毛风毛菊化学成分靶点和肾性水肿靶点，取交集获得治疗肾性水肿作用靶点;通过STRING数据库进行蛋白互作分析;利用Metascape平台进行gene ontology(GO)功能和京都基因和基因组百科全书(kyoto encyclopedia of genes and genomes，KEGG)通路分析;将药物、靶点等导入到Cytoscape3。7。2构建"药物-成分-靶点-疾病-通路"网络图。筛选出排名靠前的6个基因和12个有效成分，通过分子对接技术验证网络药理预测的准确性，揭示长毛风毛菊治疗肾性水肿的作用机制。结果 大鼠血清中TP、HDL-C指标上调，BUN、SCr、TC、TG和LDL-C指标均下调。从藏药长毛风毛菊提取物中共鉴定了 71个化合物，包括7个苯丙素类、16个绿原酸类和39个黄酮类等成分。通过预测得出75个长毛风毛菊治疗肾性水肿靶点，GO富集共涉及生物过程、细胞组分、分子功能3个方面，KEGG富集结果显示PI3K/AKT、AGE-RAGE、MAPK等是显著通路。分子对接结果显示，有效成分与关键靶点结合能Vina评分均小于0，高效验证了网络药理预测的准确性。结论 该研究基于药效学实验，结合UPLC-Q-TOF-MS/MS鉴定、网络药理学分析和分子对接技术，表明长毛风毛菊能够治疗肾性水肿，可能通过调控PI3K/AKT、AGE-RAGE、MAPK信号通路中TP53、PI3K、AKT等相关靶点来发挥作用。
Study on the chemical constituents and mechanism of Saussurea hyracoids Hook.f.extract in the treatment of renal edema
Objective To illuminate the pharmacodynamics and chemical constituents of Tibetan medicine Saussurea hieracioides Hook.f.,explore their effective components,key targets and related pathways,and to clarify its mechanism in the treatment of renal edema;Methods SD rats were assigned into blank group,doxorubicin-induced renal edema model group,low-dose group and high-dose group.The serum biochemical indexes were measured,and the indexes of total protein(TP),blood urea nitrogen(BUN),serum creatinine(SCr),triglyceride(TG),serum total cholesterol(TC),low density lipoprotein(LDL-C)and high density lipoprotein(HDL-C)in each group were discussed.To analyze the chemical components of Tibetan medicine Saussurea hieracioides Hook.f.by UPLC-Q-TOF-MS/MS technology.The chemical component targets and renal edema targets of Saussurea hieracioides Hook.f.were obtained in SwissTargetPrediction and GeneCards databases,respectively,and the common targets were selected as the targets in the treatment of renal edema.Protein-protein interaction was analyzed via STRING.Gene ontology(GO)annotation and kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment were carried out via Metascape.The drugs and targets were imported into Cytoscape 3.7.2 to construct the"drug-component-target-disease-pathway"network diagram.The top-ranked 6 genes and 12 effect components were selected,and the accuracy of network pharmacological prediction was verified by molecular docking technology,to reveal the mechanism of Saussurea hieracioides Hook.f.in the treatment of renal edema.Results The indexes of TP and HDL-C in serum of rats were up-regulated,and the indexes of BUN,SCr,TC,TG and LDL-C were down-regulated.A total of 71 compounds,including 7 phenylpropanoids,16 chlorogenic acids and 39 flavonoids,were identified from the extract of Tibetan medicine Saussurea hieracioides Hook.f..A total of 75 targets for the treatment of renal edema were predicted.GO enrichment involved three aspects:biological process(BP),cell components(CC)and molecular function(MF).The enrichment results of Kyoto Encyclopedia of Genes and Genomes(KEGG)showed that PI3K/AKT,AGE-RAGE and MAPK were significant pathways.The results of molecular docking showed that the binding energy Vina scores of effect components and key targets were less than 0,which effectively verified the accuracy of network pharmacological prediction.Conclusion Based on pharmacodynamic experiments,combined with UPLC-Q-TOF-MS/MS identification,network pharmacology analysis and molecular docking technology,this study indicating that Saussurea hieracioides Hook.f.had a significant effect on renal edema,which may play a role by regulating TP53,PI3K,AKT and other related targets in PI3K/AKT,AGE-RAGE,MAPK signaling pathway.
万方数据
维普
