基于网络药理学和分子对接探讨黄芪治疗特发性肺纤维化的分子机制

Study on the molecular mechanism of Astragali Radix in treating idiopathic pulmonary fibrosis based on network pharmacology and molecular docking

本研究旨在以黄芪活性成分为切入点,基于网络药理学及分子对接研究黄芪治疗特发性肺纤维化的分子机制。首先,通过TCMSP筛选黄芪的活性成分;利用Swiss Target Prediction预测黄芪化学成分潜在靶点;使用GeneCards和CTD筛选出特发性肺纤维化的相关基因,交集获得黄芪治疗特发性肺纤维化的潜在靶点,对潜在靶点进行生物信息学分析明确关键靶点。然后,通过采用分子对接(SYBYL 2.1.1)验证关键靶点与黄芪化学成分的结合程度。最后通过HE染色、Masson染色及ELISA实验验证网络药理学富集分析结果。最终,通过网络药理学初步筛选得到黄芪治疗特发性肺纤维化的信号通路29条(P<0.05),靶点25个,其中IL-17信号通路、EGFR信号通路和HIF-1信号通路为疾病相关通路,PTGS2、VEGFA、MMP-9、STAT3和EGFR为关键靶点;通过分子对接明确黄芪中6个化学成分与5个关键靶点均有较好的结合;HE染色和Masson染色均提示黄芪及其活性成分叶酸对博来霉素诱导的大鼠特发性肺纤维化具有治疗作用,可降低大鼠特发性肺纤维化程度;ELISA实验证明黄芪及其活性成分叶酸能够降低大鼠血清IL-17、MMP-9的表达。本研究通过网络药理学、分子对接及实验验证结果可以明确黄芪治疗特发性肺纤维化的关键靶点及主要化学成分,为开发黄芪抗特发性肺纤维化的有效成分及作用机制提供新的思路方法,为临床有效应用黄芪治疗特发性肺纤维化提供理论依据。

To study the molecular mechanism of Astragali Radix for idiopathic pulmonary fibrosis,we used the active ingredients of Astragali Radix as an entry point,based on network pharmacology and molecular docking.Firstly,the active ingredients of Astragali Radix were screened by TCMSP.The potential targets of Astragali Radix chemical components were predicted by Swiss Target Prediction.GeneCards and CTD were used to screen out the related genes of idiopathic pulmonary fibrosis,and obtain the potential targets of Astragali Radix for idiopathic pulmonary fibrosis.Bioinformatics analysis of the potential targets were conducted to clarify the key targets.Secondly,molecular docking(SYBYL 2.1.1)was used to verify the degree of binding between key targets and the chemical components of Astragali Radix.Then,HE staining,Masson staining and ELISA were used to verify the results of network pharmacological enrichment analysis.Finally,through network pharmacology preliminary screening,29 signal pathways and 25 targets of Astragali Radix were obtained for idiopathic pulmonary fibrosis,among which the IL-17 signal pathway,EGFR signal pathway and HIF-1 signal pathway were disease-related pathways.And PTGS2,VEGFA,MMP-9,STAT3 and EGFR were the key targets.Through molecular docking,we found that the six chemical components in Astragali Radix had good combination with the five key targets.Both HE staining and Masson staining suggested that Astragali Radix and its active ingredient folic acid had therapeutic effect on bleomycin-induced idiopathic pulmonary fibrosis in rats,and could reduce the degree of idiopathic pulmonary fibrosis in rats.ELISA showed that Astragali Radix and its active ingredient folic acid could reduce the expression of IL-17 and MMP-9 in serum of rats.This study clarified the key targets and main chemical components of Astragali Radix for idiopathic pulmonary fibrosis through network pharmacology,molecular docking and experimental verification,and provides new ideas and methods for the development of the effective components and mechanism of Astragali Radix for idiopathic pulmonary fibrosis and provides a theoretical basis for the clinical application of Astragali Radix for idiopathic pulmonary fibrosis.