基因测序联合网络药理学探讨补肾通蚀丸治疗酒精性股骨头坏死的作用机制  

Exploration of the mechanism of Bushen Tongshi pill in the treatment alcoholic femoral head necrosis by gene sequencing and network pharmacology

作　　者：陈财 曾平[2] CHEN Cai;ZENG Ping

机构地区：[1]广西中医药大学,广西南宁530299 [2]广西中医药大学第一附属医院,广西南宁530023

出　　处：《中医临床研究》2024年第18期115-122,共8页Clinical Journal Of Chinese Medicine

基　　金：国家自然科学基金项目:基于METTL3调控HIST1H3F的m6A甲基化修饰探讨三七总皂苷在酒精性股骨头坏死中的保护作用与机制,基金编号:82160913,项目负责人:曾平。

摘　　要：目的:通过网络药理学联合基因测序技术探讨补肾通蚀丸治疗酒精性股骨头坏死的潜在分子作用机制。方法:从中药系统药理数据库和分析平台(TCMSP)与BATMAN数据库中寻找补肾通蚀丸组成中药的活性有效成分和作用靶点。提取3名酒精性股骨头坏死患者和3名健康志愿者外周血进行基因测序,利用R语言软件中的“DESeq2”包筛选具有差异表达的信使核糖核酸(Msessenger RNA,mRNA)。通过perl软件将基因测序获得的差异基因与药物靶基因取交集,剔除重复有效成分,获得本研究的关键基因及其对应的有效成分。将关键基因及其对应的有效成分导入Cytoscape3.8.0软件中构建成分-靶点网络。利用STRING在线数据库和R语言软件筛选出度(Degree)值得分前10的关键基因。对获得的关键基因进行基因本体论(GO)功能富集和京都基因与基因组百科全书(KEGG)通路富集分析。Degree值前2的目标基因与其对应的有效成分进行分子对接验证。结果:基因测序共获得921个差异表达基因。补肾通蚀丸共获得483个药物靶基因,74个不重复有效成分,两者交集共获得27个关键基因。半胱氨酸蛋白酶(Caspase,CASP)9和胰岛素样生长因子结合蛋白3(Insulin Like Growth Factor Binding Protein 3,IGFBP3)是本次实验获得的目标基因,β-谷甾醇、槲皮素、β-胡萝卜素、木犀草素和川陈皮素是其对应的有效成分。GO结果表明关键基因生物学过程涉及白细胞凋亡过程、血小板活化和对皮质类固醇的反应;KEGG结果表明关键基因富集在卵巢类固醇生成信号通路。分子对接显示CASP9和IGFBP3与其对应的有效成分结合良好。结论:补肾通蚀丸是通过β-谷甾醇、槲皮素、木犀草素和川陈皮素等有效成分,抑制CASP9和IGFBP3靶点蛋白的表达,调控卵巢类固醇生成通路,参与骨代谢,调节骨稳态,促进血管生成和抑制炎症因子表达,发挥治疗酒精性股骨头坏死的�OObjective:To explore the potential molecular mechanism of Bushen Tongshi pill(补肾通蚀丸)in the treatment of alcoholic femoral head necrosis by network pharmacology and gene sequencing.Methods:The active ingredients and action targets of Bushen Tongshi pill were found from TCM System Pharmacological Database and Analysis Platform(TCMSP)and BATMAN database.Peripheral blood of 3 patients with alcoholic femoral head necrosis and 3 healthy volunteers were extracted for gene sequencing,and the differentially expressed mRNA was screened by using the“DESeq2”package in R language software.Through Perl software,the differential genes obtained by gene sequencing were intersected with drug target genes,and the duplicate active components were removed to obtain the key genes and their corresponding active components in this study.Key genes and their corresponding active components were introduced into Cytoscape3.8.0 software to construct the component-target network.10 key genes of degree score were screened using STRING online database and R language software.The key genes were enriched by GO function and KEGG pathway.The target genes with the top 2 degrees were verified by molecular docking with their corresponding active components.Results:A total of 921 differentially expressed genes were obtained by gene sequencing.A total of 483 target genes and 74 non-repetitive active components were obtained from Bushen Tongshi pill,and 27 key genes were obtained from their intersection.CASP9 and IGFBP3 were the target genes obtained in this experiment,andβ-sitosterol,quercetin,β-carotene,luteolin and tangerine were the corresponding effective components.GO results showed that the key gene biological processes involved leukocyte apoptosis,platelet activation and response to corticosteroids.KEGG results showed that key genes were enriched in the ovarian steroidogenic signaling pathway.Molecular docking showed that CASP9 and IGFBP3 were well bound to their corresponding active components.Conclusion:Bushen Tongshi pill inhi

关 键 词：酒精性股骨头坏死 补肾通蚀丸 基因测序 网络药理学 

分 类 号：R322.72[医药卫生—人体解剖和组织胚胎学]