基于分子对接技术与实验验证探究熟地锁阳方治疗特发性肺纤维化的疗效及其作用靶点  

The Therapeutic Effect and Targets of Shudi Suoyang Formula(熟地锁阳方)on Idiopathic Pulmonary Fibrosis were Preliminarily Explored Based on the Molecular Docking Technology and Experimental Verification

作　　者：张宁 郭亚丽[1] 彭艳茹[1,2] 王玉光 ZHANG Ning;GUO Yali;PENG Yanru;WANG Yuguang(Beijing Hospital of Traditional Chinese Medicine,Capital Medical University,Beijing 100010,China;Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]首都医科大学附属北京中医医院,北京100010 [2]北京中医药大学,北京100029

出　　处：《中医药导报》2023年第8期19-25,共7页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：首都卫生发展科研专项项目(首发2020-2-2233)。

摘　　要：目的:运用分子对接技术与动物实验验证相结合的方法,对熟地锁阳方治疗特发性肺纤维化(IPF)的疗效及其可能的作用靶点进行探究。方法:应用TCMSP结合BATMAN-TCM数据库收集熟地锁阳方的有效成分及其作用靶点,通过Degree评分选取分数较高的化合物作为配体进行分子对接。应用GeneCards、PharmGkb及OMIM等数据库获得疾病靶基因,将药物潜在作用靶基因与疾病靶基因取交集,利用CytoNCA进行拓扑分析,筛选获得核心作用靶点。根据核心靶点及文献研究选取相关靶点作为大分子受体,应用Autodock-Vina软件进行分子对接。同时建立肺纤维化小鼠模型,通过体内动物实验进行验证,运用EMMS eSpira Forced Maneuvers有创肺功能仪检测小鼠的用力肺活量(FVC)、吸气能力(IC),收集小鼠的肺组织、称重并计算肺系数,分别运用HE和Masson法观察小鼠的肺组织病理学变化,Western blotting法检测α-SMA蛋白相对表达量,实时荧光定量PCR检测VEGFA mRNA、EGFR mRNA相对表达量。结果:共获得115个有效化合物,Degree≥110的化合物共有5个,共筛选得到VEGFA、EGFR、MAPK1、TP53、AKTI、MAPK14、TNF、FOS、JUN 9个核心作用靶点。分子对接结果显示,VEGFA、EGFR与上述化合物均存在结合活性,且绝大部分靶点与有效化合物的结合活性较强。动物实验结果表明,与对照组比较,模型组小鼠肺功能FVC、IC均明显降低(P<0.01),肺系数及肺组织中α-SMA蛋白相对表达量均明显升高(P<0.01),肺组织病理学显示结构明显被破坏、炎症细胞渗出及胶原沉积增多,肺组织中VEGFA mRNA、EGFR mRNA相对表达量均明显升高(P<0.01);与模型组比较,熟地锁阳方组小鼠肺功能FVC、IC均明显升高(P<0.01),肺系数及肺组织中α-SMA蛋白相对表达量均明显降低(P<0.01),肺组织结构改善明显、炎症细胞渗出及胶原纤维沉积明显减少,肺组织中VEGFA mRNA、EGFR mRNA相对表达量均明显降低(P<0.01)。�Objective:To explore the efficacy and possible targets of Shudi Suoyang Formula in the treatment of idiopathic pulmonary fibrosis(IPF)by using molecular docking technology and animal experimental verification.Methods:TCMSP combined with BATMAN-TCM database was used to collect the effective components and targets of Shudi Suoyang Formula,and compounds with higher scores were selected as ligands for molecular docking by Degree score.GeneCards,PharmGkb and OMIM databases were used to obtain disease target genes.The potential target genes of drugs were intersected with disease target genes,and CytoNCA was used for topological analysis to screen out the core targets.According to the core target and literature research,relevant targets were selected as macromolecular receptors,and Autodock-Vina software was used for molecular docking.At the same time,a mouse model of pulmonary fibrosis was established and verified by in vivo animal experiments.The forced vital capacity(FVC)and inspiratory capacity(IC)of mice were detected by EMMS eSpira Forced Maneuvers invasive lung function instrument.The lung tissue of mice was collected,weighed and the lung coefficient was calculated.The pathological changes of lung tissue in mice were observed by HE and Masson methods,and the relative expression ofα-SMA protein was detected by Western blotting.Real-time fluorescence quantitative PCR was used to detect the relative expression of VEGFA mRNA and EGFR mRNA.Results:A total of 115 effective compounds were obtained,and 5 compounds with Degree≥110 were obtained.A total of 9 core targets of VEGFA,EGFR,MAPK1,TP53,AKTI,MAPK14,TNF,FOS and JUN were screened.The results of molecular docking showed that VEGFA and EGFR had binding activity with the above compounds,and most of the targets had strong binding activity with the effective compounds.The results of animal experiments showed that compared with the control group,the FVC and IC of the lung function in the model group were significantly decreased(P<0.01),the lung coefficient and the rela

关 键 词：特发性肺纤维化 熟地锁阳方 分子对接技术 疗效 作用靶点 小鼠 

分 类 号：R285.5[医药卫生—中药学]