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    基于网络药理学和实验验证的三黄膏治疗痔疮的作用机制研究

    Mechanism of Sanhuang Ointment in Treatment of Haemorrhoids Based on Network Pharmacology and Experimental Verification

      摘要
    • 摘要:
      目的 基于网络药理学对三黄膏治疗痔疮的核心成分、靶点及信号通路进行预测，并借助分子对接和实验验证，对其作用机制进行初步探讨。
      方法 经过文献研究和数据库筛选，得到三黄膏活性成分、靶点及痔疮相关靶点，通过Cytoscape软件构建关联网络并筛选核心基因，然后对基因进行功能分析和通路富集分析。使用Autodock Vina进行分子对接验证，并通过酶联免疫吸附法(ELISA)和免疫组化验证三黄膏对核心靶点的调控作用。
      结果 收集到三黄膏治疗痔疮活性成分77个及其潜在靶点61个，KEGG显示三黄膏可调控钙信号通路、神经活性配体-受体相互作用、cGMP-PKG信号通路等，影响CGRP、NK-1R、TRPV1、VEGF、IL-6和TNF-α 6个核心靶点，分子对接显示三黄膏与靶点蛋白间有较好的亲和性，动物实验验证了三黄膏可抑制IL-6、CGRP、P物质等的表达，促进VEGF的表达。
      结论 三黄膏可能作用于CGRP、NK-1R等靶点，通过调控钙信号通路及PI3K-Akt等通路，发挥镇痛抗炎、活血化瘀作用来治疗痔疮。

       

      Abstract:
      OBJECTIVE To predict the core ingredients, targets and signaling pathways of Sanhuang ointment in the treatment of haemorrhoids based on network pharmacology, and preliminarily discuss the mechanism of action preliminarily by means of molecular docking and experimental verification.
      METHODS Active ingredients, targets and haemorrhoid related targets of Sanhuang ointment were obtained through literature research and database screening. Association networks were constructed and core targets were screened by Cytoscape, and gene function analysis and pathway enrichment analysis were conducted. Autodock Vina was used for molecular docking verification, and the regulatory effects of Sanhuang ointment on core targets were verified by ELISA and immunohistochemistry.
      RESULTS The 77 active ingredients and 61 potential targets of Sanhuang ointment for treating haemorrhoids were collected. KEGG showed that Sanhuang ointment could regulate calcium signaling pathways, neuroactive ligand receptor interactions, cGMP-PKG pathway, etc. Affect 6 core targets of CGRP, NK-1R, TRPV1, VEGF, IL-6 and TNF-α. Molecular docking showed that Sanhuang ointment had a good affinity with target protein, and animal experiments verified that Sanhuang ointment could inhibit the expression of IL-6, CGRP, substance P, and promote the expression of VEGF.
      CONCLUSION Sanhuang ointment may act on CGRP, NK-1R and other targets, regulate Calcium signaling pathway and PI3K-Akt pathways, and exert analgesic and anti-inflammatory effects, promoting blood circulation and removing blood stasis to treat haemorrhoids.

       

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