| 霍艳杰,何超平,全文娟,陈煜,庹勤慧.基于网络药理学初探川续断皂苷乙的药理作用.[J].中南医学科学杂志.,2021,(1):51-57. |
| 基于网络药理学初探川续断皂苷乙的药理作用 |
| Study on the pharmacological action of dipsacoside B by network pharmacology |
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| DOI：10.15972/j.cnki.43-1509/r.2021.01.010 |
| 中文关键词: 川续断皂苷乙 网络药理学 拓扑异构酶2a 血管平滑肌细胞增殖 动脉粥样硬化 |
| 英文关键词:dipsacoside B network pharmacology TOP2a VSMCs proliferation atherosclerosis |
| 基金项目: |
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| 摘要点击次数: 596 |
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| 中文摘要: |
| 目的利用网络药理学与分子生物学方法探究川续断皂苷乙(DB)的药理作用。 方法在ChemMapper数据库上预测DB的靶标,通过iGEMDOCK软件进行化合物与靶标蛋白对接,根据评分选出关键靶标蛋白,关键靶标蛋白通过STRING数据库构建蛋白之间的相互作用,GO富集分析与KEGG通路分析关键靶标蛋白调控蛋白网络的生物过程与通路。血管紧张素Ⅱ(AngⅡ)诱导血管平滑肌细胞(VSMCs)增殖,通过MTT法与EdU法检测DB对VSMCs的细胞活性及增殖的影响。流式细胞术检测VSMCs周期,Western blot法与免疫荧光法检测关键靶标蛋白表达。 结果DB具有4个靶标蛋白,根据分子对接能量值选取DNA拓扑异构酶2a(TOP2a)后续研究对象,TOP2a相关的蛋白进行预测,得到相关蛋白10种,经过GO富集分析与KEGG通路分析,TOP2a可能参与VSMCs的增殖调控。MTT法结果显示DB对VSMCs活性明显抑制,具有剂量依赖性。EdU法检测DB(3、10、30 μmol/L)对VSMCs增殖有明显抑制作用。流式细胞术显示DB可能阻滞VSMCs的G1期向S期合成,进而影响细胞增殖。Western blot法结果显示DB下调蛋白TOP2a表达水平,同样免疫荧光法结果显示TOP2a表达下调。 结论DB可能通过下调TOP2a表达,调控细胞周期,抑制VSMCs的增殖。 |
| 英文摘要: |
| To explore the pharmacological effects of dipsacoside B (DB) via network pharmacology and molecular biology. MethodsChemMapper database was used to predict the target proteins of DB. The compounds were docked with the target proteins by iGEMDOCK software, and the key target proteins were selected according to the score. STRING database was constructed to analyze the interaction of key target proteins with others. GO enrichment analysis and KEGG pathway analysis were used to analyze the regulatory effects of key target protein on the biological processes and pathways of the protein network. Proliferation model of vascular smooth muscle cells (VSMCs) was induced by angiotensinogen Ⅱ (AngⅡ). MTT and EdU methods were used to detect the effects of DB on the activity and proliferation of VSMCs. The cell cycle of VSMCs was measured by flow cytometry, and the expression of key target proteins was determined by western blot and immunofluorescence. ResultsDB had four target proteins, while DNA topisomerase 2a (TOP2a) was selected for further study according to the score and 10 related proteins were obtained. MTT results showed that DB presented a dominant inhibitory effect on VSMCs activity in a dosage-depend manner. EdU results showed that the significant inhibition dosage was at 3,10, 30 μmol/L. Flow cytometry results showed that DB can block the synthesis of VSMCs throughout G1 phase to S phase, thus affecting cell proliferation. Western blot and immunofluorescence results both showed that DB down-regulated the protein expression of TOP2a. ConclusionDB may inhibit the proliferation of VSMCs by down-regulating the protein expression of TOP2a and regulating cell cycle. |
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