【作者】 李健；

【导师】 郭玉海；

【作者基本信息】 广州中医药大学 ， 中医骨伤科学（专业学位）， 2023， 硕士

【摘要】 目的:运用数据挖掘技术研究现代医家治疗气滞血瘀型腰椎间盘突出症(lumbar disc herniation,LDH)的用药规律,使用相关算法建立中药治疗气滞血瘀型LDH的核心药物组合,并对该药物组合展开网络药理学分析,预测药物活性成分发挥疗效的相关药理学机制。为临床治疗气滞血瘀型LDH提供新的诊疗思路,同时进一步优化中药内服治疗气滞血瘀型LDH的诊疗方案。方法:利用文献检索的方法,在中国知网(China national knowledge internet,CNKI)、万方、维普中文期刊数据库搜索自建库以来至2023年2月10日发表的关于中医药方剂内服治疗气滞血瘀型LDH的临床试验文献。对筛选后得到的文献进行方剂名称、药物组成、药物归经及功效分类等数据进行统计及规范化处理。将所得到的数据导入SPSS软件进行频次分析、聚类分析及关联规则分析,挖掘出核心药物组合。在TCMSP数据库获取药物活性成分及靶点;通过Gene Cards疾病数据库获取LDH相关靶点,筛选出疾病与药物的共同靶点。使用Cytoscape软件构建“中药—成分—靶点网络图”并寻找核心药物组合治疗气滞血瘀型LDH的关键中药成分。利用STRING平台构建PPI网络并筛选出关键靶点。将共有靶点上传至 DAVID 数据库(database for annotation,visualization and integrated discovery,DAVID)进行GO功能富集分析和KEGG通路富集分析。设置P<0.05,分别选取排名前10的结果,导入微生信在线网站进行可视化。将关键靶点蛋白与其对应中药活性成分导入AutoDock Vina软件进行分子对接。结果:检索到文献525篇,通过阅读文献题目及摘要初步筛选出文献109篇,根据文献纳排标准精读文献全文后最终纳入文献101篇,共收录处方104首。共涉及中药103味,累计使用频次达1279次。药物频次排名前十位的分别是当归、川.芎、红花、桃仁、甘草、地龙、牛膝、没药、香附、秦艽。共涉及17个中药功效分类,其中用药频次排名前五的分别是活血化瘀药、补虚药、祛风湿药、理气药、平肝熄风药。所有归经总频次为3219次,药物归经频数排名前五的分别:肝经、脾经、心经、肾经、胃经。高频药物聚类分析得到三类药物,第一类:当归、川芎、红花、桃仁、甘草、地龙、牛膝、没药、香附、秦艽、五灵脂、羌活;第二类:桑寄生、续断、土鳖虫、赤芍、黄芪、杜仲;第三类:白芍、延胡索。关联规则分析得到置信度和规则支持度排名前三的药物组合为:香附一当归、牛膝—桃仁—红花、牛膝—桃仁—川芎—红花。结合关联规则分析结果与中药使用频次统计情况,挖掘到临床治疗气滞血瘀型LDH核心药物组合为:香附、当归、牛膝、川芎、桃仁、红花。在TCMSP数据库中共检索到核心药物组合相关活性成分77个,筛选出关键活性成分10个,包括槲皮素、木犀草素、β-谷甾醇、山柰酚、黄芩素、β-胡萝卜素、豆甾醇、异鼠李素、金圣草黄素、汉黄芩素。检索到核心药物组合活性成分相关靶点208个,LDH相关靶点545个,两者共同靶点55个。构建PPI网络筛选出关键靶点为IL6、IL1B、ALB、AKTl、MMP9、VEGFA。KEGG分析结果共得到相关信号通路125条,主要包括IL-17信号通路、AGE-RAGE信号通路、TNF信号通路、PI3K-Akt信号通路、MAPK信号通路等。分子对接结果显示,关键靶点与其对应化合物具有强烈的结合活性。结论:本研究结果显示,中药治疗气滞血瘀型LDH的常用方剂为身痛逐瘀汤。使用频率较高的中药包括当归、川芎、红花、桃仁、甘草、地龙、牛膝、没药、香附、秦艽等;常用的药物组合包括香附一当归、牛膝一桃仁—红花、牛膝—桃仁—川芎—红花等。结合关联规则分析结果与中药使用频次统计情况,本研究得到临床治疗气滞血瘀型LDH的核心药物组合为香附、当归、牛膝、川芎、桃仁、红花。网络药理学研究发现,香附、当归、牛膝、川弯、桃仁、红花治疗气滞血瘀型LDH的主要活性成分包括槲皮素、木犀草素、β-谷甾醇、山柰酚、黄芩素、β-胡萝卜素、豆甾醇、异鼠李素、金圣草黄素、汉黄芩素等;可能通过作用于IL6、IL1B、ALB、AKT1、MMP9、VEGFA 等关键靶点蛋白,调控 IL-17、AGE-RAGE、TNF、PI3K-Akt、MAPK等信号通路,从调节细胞凋亡、降低炎症水平、调节机体免疫反应、延缓椎间盘退行性变等多个方面治疗气滞血瘀型LDH。同时也反映出香附、当归、牛膝、川芎、桃仁、红花治疗气滞血瘀型LDH是一个涉及多成分、多靶点、多通路的复杂生物学过程。更多还原

【Abstract】 ObjectiveData mining technology was used to study the medication rules of modern doctors in the treatment of lumbar disc herniation(LDH)of qi stagnation and blood stasis type.Related algorithms were used to establish the core drug combination of traditional Chinese medicine for the treatment of LDH of qi stagnation and blood stasis type,and the core drugs were analyzed by the method of network pharmacology.To predict the pharmacological mechanisms underlying the therapeutic effects of drug active ingredients.It provides a new diagnosis and treatment idea for clinical treatment of LDH of qi stagnation and blood stasis type,and further optimizates the diagnosis and treatment plan of oral Chinese medicine for LDH of qi stagnation and blood stasis type.MethodsLiterature retrieval method was used to search the clinical trials of LDH of qi stagnation and blood stasis type with traditional Chinese medicine formula published in the Chinanational knowledge internet(CNKI),Wanfang and VIP database from the establishment of the database to February 10,2023.After screening,the data of prescription name,drug composition,drug attribution and efficacy classification were statistically analyzed and standardized.The obtained data were imported into SPSS software for frequency analysis,cluster analysis and association rule analysis.The active ingredients and targets of the drug were obtained from the TCMSP database.LDH-related targets were obtained through Gene Cards disease database,and the common targets of diseases and drugs were screened.Cytoscape software was used to construct the "TCM components-target network diagram" and find the key TCM components of core drug combination in the treatment of LDH of qi stagnation and blood stasis type.The STRING platform was used to construct the PPI network and screen out the key targets.The consensus targets were uploaded to the database for annotation,visualization and integrated discovery(DAVID)for GO functional enrichment analysis and KEGG pathway enrichment analysis.Set P<0.05,and select the top 10 results,respectively,and import them into the wechat online website for visualization.Key target proteins were imported into AutoDock Vina software for molecular docking.ResultsA total of 525 Chinese literatures were retrieved,109 literatures were preliminarily screened by reading the title and abstract of the literature,and 101 literatures were finally included after reading the full text of the literature according to the literature inclusion and exclusion criteria,including 104 prescriptions.A total of 103 Chinese herbs were involved,with a total frequency of 1279 times.The top ten drugs were Angelica sinensis,Ligusticum chuanxiong,Safflower,peach seed,Glycyrrhizae radix,Dipterocarpus vulgaris,Achyranthes bidentata,myrrha,Radix gentiana gentiana.A total of 17 TCM efficacy categories were involved,among which the top five medication frequency were invigorating blood stasis,tonifying deficiency,dispelling wind and dampness,regulating qi,and soothing liver and extinguishing wind.The total frequency of all the meridians was 3219 times,and the top five in the frequency of drug meridians were liver meridian,spleen meridian,heart meridian,kidney meridian and stomach meridian.Three kinds of drugs were found by cluster analysis of high frequency drugs.The first type was Angelica sinensis,Ligusticum chuanxiong,Safflower,peach kernel,Licorice,Lumbricus,Achyrantha bidentata,myrrha,Xiangfu,Gentiana gentiana,Wulingzhi and Qianghuo.The second category:mulberry parasitics,Dipsacus,Earthworms,red Peony,Astragalus,Eucommia;The third category:Radix paeoniae and Rhizoma corydalis.The top three drug combinations with high confidence and high rule support were Xiangfu-Danggui(Angelica sinensis),Niuxi-Taoren-Safflower,Niuxi-taoren-Chuangxiongsafflower.Combined with the results of association rule analysis and the statistics of the frequency of traditional Chinese medicine use,the core drug combinations for clinical treatment of LDH of qi stagnation and blood stasis type were found to be:Xiangfu,Angelica sinensis,Niuxi,Chuangxiong,peach kernel,safflower.A total of 77 active ingredients related to the core drug combination were retrieved from the TCMSP database,and 10 key active ingredients were screened,including quercetin,luteolin,β-sitosterol,kaempferol,baicalein,β-carotene,stigmasterol,isorhamnetin,eriocanthin,and wogonin.208 targets related to the active ingredients of the core drug combination,545 targets related to LDH,and 55 common targets of the two were retrieved.The key targets of PPI network were IL6,IL1B ALB,AKT1,MMP9 and VEGFA.A total of 125 related signaling pathways were obtained by KEGG analysis,mainly including IL-17 signaling pathway,AGE-RAGE signaling pathway,TNF signaling pathway,PI3K-Akt signaling pathway,MAPK signaling pathway,etc.The results of molecular docking showed that the key target proteins had strong binding activity with their corresponding compounds.ConclusionThe results of this study showed that Shentong Zhuyu decoction was the most common prescription for the treatment of LDH of qi stagnation and blood stasis type.The most frequently used herbs were Angelica sinensis,Ligusticum chuanxiong,Safflower,peach seed,Glycyrrhizae radix,Pteronyssinus,Achyranthes bidentata,myrrha,Radix gentiana gentiana,etc.The commonly used drug combinations include Xiangfu-Angelica sinensis,Niuxi-peach kernel-safflower,Niuxi-peach kernelChuanxiong-safflower,etc.Combined with the results of association rule analysis and the frequency statistics of traditional Chinese medicine use,this study obtained that the core drug combinations for clinical treatment of LDH of qi stagnation and blood stasis type were Xiangfu,Angelica sinensis,Niuxi,Chuanxiong,peach kernel and safflower.Network pharmacology study found that the main active ingredients of Xiangfu,Angelica sinensis,Achyrantha bidentata,Ligusticum,peach seed and safflower in the treatment of Qi-stasis and blood-stasis type LDH included quercetin,luteolin,β-sitosterol,kaempferol,baicalein,β-carotene,stigmasterol,isorhamnetin,eriocanthin,wogonin,etc.It may regulate IL-17,AGE-RAGE,TNF,PI3K-Akt,MAPK and other signaling pathways by acting on IL6,IL1B,ALB,AKT1,MMP9,VEGFA and other key target proteins.LDH of qi stagnation and blood stasis type can be treated from many aspects such as regulating cell apoptosis,reducing inflammation level,regulating the body’s immune response,and delaying intervertebral disc degeneration.At the same time,it also reflects that the treatment of qi stagnation and blood stasis type LDH with Xiangfu,Angelica,Niuxi,Chuangxiong,peach kernel and safflower is a complex biological process involving multiple components,multiple targets and multiple pathways.更多还原