参麦注射液治疗膝关节骨关节炎的网络药理学分析及临床对照验证

Network pharmacology analysis and clinical verification of Shenmai injection in the treatment of knee osteoarthritis

目的阐述参麦注射液治疗膝关节骨关节炎(knee osteoarthritis,KOA)的物质基础及分子机制并进行临床验证。方法通过数据库筛选有效化学成分。使用Cytoscape软件构建参麦注射液(shenmai injection,SMI)关键靶点-KOA复合目标网络。对关键靶点进行基因本体论生物功能分析及基因和基因组的京都百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。对核心关键靶点进行分子对接验证。选取2017年5月至2019年11月在我院治疗的KOA患者130例。根据随机数字表法分为观察组及对照组,每组各65例。观察组采用SMI膝关节腔内注射治疗。对照组采用透明质酸钠注射液膝关节腔内注射治疗。采用McMaster大学骨关节炎指数(the western ontario and McMaster universities,WOMAC)、健康调查简表(the MOS item short from health survey,SF-36)评估两组患者治疗后6周、12周、6个月及12个月后效果及生活质量。结果(1)共获得25个SMI抗KOA关键靶点,麦冬中麦冬酮A、鸟苷、豆甾醇、麦冬A及尿苷。红参中β-谷固醇、人参皂苷rh2是复合目标网络中主要有效化学成分。拓扑分析得到胱天蛋白酶1(caspase 1,CASP1)、CASP3、CASP8、CASP9、核因子-κB抑制因子α(nuclear factor-κB inhibitorα,NFKBIA)、雌激素受体1(estrogen receptor 1,ESR1)、过氧化氢酶(catalase,CAT)核心关键靶点。KEGG富集分析结果显示细胞凋亡(apoptosis)富集程度最高。CASP1、CASP3、CASP9、NFKBIA、ESR1、CAT与SMI有效活性成分结合能均<-5.0 kcal/mol,至少存在1个氢键。豆甾醇对接CAT存在4个氢键。(2)观察组患者中3例没有按时完成SMI腔内注射,1例随访丢失;对照组患者中1例没有按时完成透明质酸钠注射液腔内注射,1例随访丢失。治疗后6周、12周、6个月、12个月,观察组WOMAC中疼痛、僵硬、功能评分低于对照组(P<0.05);观察组SF-36中躯体健康评分高于对照组(P<0.05)。结论SMI治疗KOA具有多成分、多途径、多靶点协同干预作用特点,能明显改善KOA的临床症状及提高生活质量并延缓KOA病程进展。

Objective To explore the material basis and molecular mechanism of Shenmai injection(SMI)in the treatment of knee osteoarthritis(KOA)and to conduct clinical verification.Methods Traditional Chinese Medicine Systems Pharmacology(TCMSP)database and bioinformatics analysis tool for molecular mechanism of traditional Chinese medicine(BATMAN)database were used to screen the effective chemical constituents.Using Cytoscape software to construct SMI key target koa composite target network.Gene ontology(Go)biological function analysis and kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were performed for key targets.The key target was verified by molecular docking.From May 2017 to November 2019,130 patients with koa who were treated in our hospital were selected.All were randomly divided into observation group and control group,65 cases in each group.The observation group was treated with intra-articular injection of SMI.The control group was treated with intra-articular injection of sodium hyaluronate.McMaster University Osteoarthritis Index(WOMAC)and short form of Health Survey(SF-36)were used to evaluate the effect and quality of life after 6 weeks,12 weeks,6 months and 12 months.Results(1)A total of 25 SMI anti koa key targets were obtained,including ophiopogon a,guanosine,stigmasterol,ophiopogon A and uridine.β-sitosterol and ginsenoside Rh2 in red ginseng were the main effective components in the complex target network.The key targets of CASP1,CASP3,CASP8,CASP9,NFKBIA,ESR1 and CAT were obtained by topological analysis.KEGG enrichment analysis showed that apoptosis was the most abundant.The binding energies of CASP1,CASP3,CASP9,NFKBIA,ESR1 and CAT with SMI active components were less than-5.0 kcal/mol,and there was at least one hydrogen bond.There were 4 hydrogen bonds between stigmasterol and CAT.(2)In the observation group,3 patients did not complete the SMI intracavitary injection on time and 1 patient lost the follow-up;in the control group,1 patient did not complete the sodium hyaluronate intracavitary injection on time and 1 patient lost the follow-up.After 6 weeks,12 weeks,6 months and 12 months of treatment,scores of pain,stiffness and function in WOMAC of the observation group were lower than those of the control group(P<0.05);scores of physical health in SF-36 of the observation group were higher than those of the control group(P<0.05).Conclusions SMI realizes multi-component,multi-channel and multi-target collaborative intervention to significantly improve the clinical symptoms and quality of life,while delays the progression of KOA.