摘要
目的 利用网络药理学与动物实验探索鼠妇治疗癌性疼痛的作用机制.方法 通过检索中国知网、万方数据知识服务平台和PubMed数据库中相关文献获取鼠妇的化学成分,检索时间为建库至2023年12月.利用PubChem获得各成分的SMILES结构后,进一步利用SwissADME进行筛选,继由SwissTarget Prediction平台预测药物作用靶点;由GeneCards数据库构建癌性疼痛疾病靶点库,并与鼠妇药物靶点进行映射;构建"药物-成分-共有靶点"和蛋白质-蛋白质相互作用网络,用Cytoscape软件进行网络拓扑分析和可视化;利用R包clusterProfiler对靶点和基因进行KEGG通路富集分析.选取清洁级健康雄性C57BL/6小鼠40只,6～8周龄,体重为20～25 g.其中10只设为假手术组,30只造模.采用胫骨内肺癌细胞注射构建癌痛模型.造模成功后按照随机数字表法将其分为模型组、高剂量组(200mg/kg)组、低剂量组(50mg/kg),各10只.Western blot法检测所筛选关键靶点的蛋白表达.结果 共筛选得到483个鼠妇靶点,2 940个癌性疼痛靶点,260个药物-疾病共有靶点.白蛋白、白细胞介素-6(IL-6)、血管内皮生长因子A(VEGFA)、丝裂原活化蛋白激酶(MAPK)3、EGFR、SRC、MAPK 1和ESR1被筛选为关键基因,这些靶点主要富集于PI3K-Akt信号通路.模型组小鼠脊髓中IL-6、VEGFA、MAPK3和MAPK1蛋白表达水平高于假手术组,差异有统计学意义(P＜0.05).高剂量组脊髓中IL-6、MAPK3、VEGFA和MAPK1蛋白表达水平低于模型组,差异有统计学意义(P＜0.05),低剂量组脊髓中IL-6、MAPK3和MAPK1蛋白表达水平低于模型组,VEGFA蛋白表达水平高于模型组,差异有统计学意义(P＜0.05).高剂量组脊髓中IL-6、VEGFA、MAPK1和MAPK3蛋白表达水平低于低剂量组,差异有统计学意义(P＜0.05).结论 鼠妇对癌性疼痛的小鼠的镇痛效应,其机制可能与抑制IL-6、MAPK3、VEGFA和MAPK1表达的异常增加,调节调控PI3k/Akt信号通路有关.
Abstract
Objective To explore the mechanism of Armadillidium vulgare in the treatment of cancer pain by using network pharmacology and ani-mal experiments.Methods The chemical constituents of were obtained by searching relevant literatures from CNKI,Wanfang Data,and PubMed database,and the retrieval time was from the establishment of the database to December 2023.After the SMILES structure of each component was obtained by PubChem,SwissADME was further used for screening,and the drug target was predicted by SwissTarget Prediction platform.The can-cer pain target database was constructed from GeneCards database and mapped with the drug target of Armadillidium vulgare.The"drug-composi-tion-common target"and protein-protein interaction network was constructed,and network topology analysis and visualization with Cytoscape soft-ware were carried out;KEGG pathway enrichment analysis for targets and genes using the R-package clusterProfiler.Forty clean male C57BL/6 mice,aged 6-8 weeks and weighing 20-25 g,were selected.Of these,ten were set up as sham surgery group and 30 were moulded.The cancer pain model was constructed by injection of intratibial lung cancer cells.After successful modeling,they were divided into model group,high dose group(200 mg/kg)group,and low dose group(50 mg/kg),according to random number table method with ten in each group.Western blot assay was used to detect the protein expression of the selected key targets.Results A total of 483 mouse female targets,2 940 cancer pain targets,and 260 drug-disease common targets were identified.ALB,interleukin-6(IL-6),vascular endothelial growth factor A(VEGFA),mitogen-activated protein kinase(MAPK)3,EGFR,SRC,MAPK1,and ESR1 were selected as key genes,and these targets were mainly enriched in the PI3K-Akt signaling pathway.The levels of IL-6,VEGFA,MAPK3,and MAPK1 in the spinal cord of mice in the model group were higher than those in the sham opera-tion group,and the differences were statistically significant(P＜0.05).The expression levels of IL-6,MAPK3,VEGFA,and MAPK1 in the spinal cord of high dose group were lower than those of model group,the differences were statistically significant(P＜0.05).The expression levels of IL-6,MAPK3,and MAPK1 in the spinal cord of low-dose group were lower than those of model group,while the expression levels of VEGFA were higher than those of model group,the differences were statistically significant(P＜0.05).The protein expression levels of IL-6,VEGFA,MAPK1 and MAPK3 in spinal cord of high dose group were lower than those of lowdose group,the differences were statistically significant(P＜0.05).Conclu-sion The results of this study suggest that Armadillidium vulgare have analgesic effects on cancer pain in mice,and the mechanism may be related to inhibiting the abnormal increase of IL-6,MAPK3,VEGFA,and MAPK1 expression and regulating the PI3K/Akt signaling pathway.
基金项目
北京市科技计划"十病十药"研发项目(Z161100001816014)
NETL
NSTL
万方数据