补阳还五汤治疗腰椎间盘突出症作用机制的网络药理学研究

Mechanism of action of Buyang Huanwu Tang(补阳还五汤)for treatment of lumbar disc herniation:a net-work pharmacology study

目的:采用网络药理学方法探讨补阳还五汤治疗腰椎间盘突出症(lumbar disc herniation,LDH)的作用机制。方法:通过检索中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)和文献筛选补阳还五汤的活性成分和靶点。利用在线人类孟德尔遗传(online Mendelian inheritance in man,OMIM)数据库、Gene-Cards数据库、Disgenet数据库检索LDH的靶点,通过与补阳还五汤的靶点取交集,获取补阳还五汤治疗LDH的靶点。利用Cytoscape 3.8.0软件构建补阳还五汤治疗LDH的“活性成分-靶点”网络及靶点蛋白互作网络,筛选补阳还五汤治疗LDH的主要活性成分和关键靶点,并进行分子对接。同时利用R软件进行靶点基因GO分析和KEGG分析。结果:从TCMSP和文献中共筛选到补阳还五汤的活性成分47个、靶点265个。从OMIM数据库、GeneCard数据库、Disgenet数据库共检索到LDH的靶点392个。将补阳还五汤靶点和LDH靶点取交集后得到补阳还五汤治疗LDH的靶点46个。补阳还五汤治疗LDH的“活性成分-靶点”网络和靶点蛋白互作网络显示,槲皮素、木犀草素、山柰酚及黄芩素为补阳还五汤治疗LDH的主要活性成分,JUN原癌基因(JUN proto-oncogene,JUN)、肿瘤坏死因子(tumor necrosis factor,TNF)、AKT丝氨酸/苏氨酸激酶1(AKT serine/threonine kinase 1,AKT1)、丝裂原活化蛋白激酶1(mitogen-activated protein kinase 1,MAPK1)、肿瘤蛋白P53(tumor protein P53,TP53)、基质金属蛋白酶9(matrix metalloproteinase 9,MMP9)为补阳还五汤治疗LDH的关键靶点。补阳还五汤治疗LDH的靶点基因GO分析和KEGG分析结果显示,补阳还五汤治疗LDH的分子生物学机制主要涉及晚期糖基化终末产物(advanced glycation end product,AGE)-晚期糖基化终末产物受体(advanced glycation end product receptor,RAGE)信号通路、白细胞介素17(interleukin 17,IL-17)信号通路、TNF信号通路,参与对营养水平、氧化应激、脂多糖以及细菌起源分子的反应等生物过程。分子对接结果显示,4个主要活性成分(槲皮素、木犀草素、山柰酚、黄芩素)与6个关键靶点(JUN、TNF、AKT1、MAPK1、TP53、MMP9)均具有良好的结合能力。结论:补阳还五汤可能是通过调控AGE-RAGE、IL-17及TNF等信号通路,减轻机体氧化应激反应、抑制炎症因子表达,发挥对LDH的治疗作用;槲皮素、木犀草素、山柰酚及黄芩素可能是补阳还五汤治疗LDH的主要活性成分,JUN、TNF、AKT1、MAPK1、MMP9、TP53是对应的关键靶点。

Objective:To explore the mechanism of action of Buyang Huanwu Tang(补阳还五汤,BYHWT)in treatment of lumbar disc hermniation(LDH)by using network pharmacology approach.Methods:The active ingredients and action targets of BYHWT were screened by retrieving traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP)and articles.The action targets of LDH were searched out from online Mendelian inheritance in man(OMIM)database,GeneCards database and Disgenet database.The targets of BYHWT for treatment of LDH were obtained through overlapping the targets of LDH with BYHWT.The active ingredient-target network and protein-protein interaction(PPI)network of BYHWT for treatment of LDH were built by using Cytoscape3.8.0 software for screening the main active ingredients and key targets of BYHWT for treatment of LDH,and the molecular docking was conducted.The GO function and KEGG pathway enrichment analysis were performed on key target genes respectively by using R software to dig biological functions and pathways of the key targets.Results:Forty-seven active ingredients and 265 targets of BYHWT were screened out from TCMSP and articles,and 392 targets of LDH were obtained from OMIM database,GeneCards database and Disgenet database.Forty-six targets of BYH-WT for treatment of LDH were obtained through overlapping the targets of BYHWT with LDH.The active ingredient-target network dia-gram and PPI network diagram of BYHWT for treatment of LDH illustrated that quercetin,luteolin,kaempferol and baicalein were the main active ingredients of BYHWT for treatment of LDH,and JUN proto-oncogene(JUN),tumor necrosis factor(TNF),AKT serine/threonine kinase 1(AKT1),mitogen-activated protein kinase 1(MAPK1),tumor protein P53(TP53)and matrix metalloproteinase 9(MMP9)were the key targets of BYHWT for treatment of LDH.The results of KEGG signaling pathway enrichment analysis suggested that the molecular biological mechanism of BYHWT for treatment of LDH mainly related to advanced glycation end product(AGE)-advanced glycation end product receptor(RAGE)signaling pathway,interleukin 17(IL-17)signaling pathway and TNF signaling pathway.The results of GO func-tion enrichment analysis indicated that the biological processes of BYHWT for treatment of LDH were mainly involved in the responses to.nutrient level,oxidative stress,lipopolysaccharides and bacteria-derived molecules.The results of molecular docking showed that all of the 4 main active ingredients(quercetin,luteolin,kaempferol and baicalein)could bind well to the 6 key targets(JUN,TNF,AKT1,MAPK1,TP53 and MMP9).Conclusion:BYHWT may reduce oxidative stress response and inhibit the expression of inflammatory factors through regulating AGE-RAGE,IL-17 and TNF signaling pathways for treatment of LDH.Quercetin,luteolin,kaempferol and baicalein may be the main active ingredients and JUN,TNF,AKT1,MAPK1,MMP9 and TP53 are the corresponding key targets of BYHWT in treatment of LDH.