[1]王伟伟,欧志学,章晓云,等.补肾壮筋汤治疗膝骨关节炎作用机制的网络药理学研究[J].中医正骨,2022,34(07):6-14,26.
　WANG Weiwei,OU Zhixue,ZHANG Xiaoyun,et al.Mechanism of Bushen Zhuangjin Tang(补肾壮筋汤)in treatment of knee osteoarthritis:a network pharmacology-based study[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2022,34(07):6-14,26.
点击复制

补肾壮筋汤治疗膝骨关节炎作用机制的网络药理学研究()
分享到：

《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第34卷
期数:
2022年07期
页码:
6-14,26
栏目:
基础研究
出版日期:
2022-07-20

文章信息/Info

Title:
Mechanism of Bushen Zhuangjin Tang(补肾壮筋汤)in treatment of knee osteoarthritis:a network pharmacology-based study
作者:
王伟伟1; 欧志学2; 章晓云1; 李统1
(1.广西中医药大学附属瑞康医院,广西 南宁 530011; 2.桂林市中医医院,广西 桂林 541002)
Author(s):
WANG Weiwei1; OU Zhixue2; ZHANG Xiaoyun1; LI Tong1
1.Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530011,Guangxi,China 2.Guilin Municipal Hospital of Traditional Chinese Medicine,Guilin 541002,Guangxi,China
关键词:
骨关节炎; 膝;  补肾壮筋汤;  网络药理学;  药物作用机制
Keywords:
osteoarthritis; knee;  Bushen Zhuangjin Tang;  network pharmacology;  mechanism of drug action
摘要:
目的:采用网络药理学方法探讨补肾壮筋汤治疗膝骨关节炎(knee osteoarthritis,KOA)的作用机制。方法:使用中药系统药理学数据库与分析平台、中药分子机制的生物信息学分析工具,检索和筛选补肾壮筋汤的活性成分,然后借助PubChem和Swiss TargetPrediction数据库检索和筛选活性成分对应的靶点,预测补肾壮筋汤的作用靶点。在GeneCard数据库中检索KOA的靶点。将检索到的KOA靶点和补肾壮筋汤靶点取交集,获取补肾壮筋汤治疗KOA的作用靶点。通过String数据库和Cytoscape3.7.2软件分析补肾壮筋汤治疗KOA的作用靶点,构建补肾壮筋汤治疗KOA靶点蛋白互作网络,筛选补肾壮筋汤治疗KOA的关键靶点。筛选与补肾壮筋汤治疗KOA的作用靶点有关的活性成分,并将其与补肾壮筋汤治疗KOA的作用靶点、补肾壮筋汤组成中药一同输入Cytoscape3.7.2软件,构建补肾壮筋汤治疗KOA“中药-活性成分-靶点”网络,分析补肾壮筋汤治疗KOA的主要活性成分。利用DAVID数据库和R3.6.2软件进行补肾壮筋汤治疗KOA靶点基因GO富集分析和KEGG富集分析。运用Autodock vina软件将补肾壮筋汤治疗KOA的主要活性成分与关键靶点进行分子对接,对其结合能进行评估,并通过Pymol软件将结果可视化。结果:共筛选到补肾壮筋汤的活性成分99个、预测到对应的靶点768个,通过与检索到的989个KOA靶点取交集,共获得补肾壮筋汤治疗KOA的靶点172个。通过分析共获得15个补肾壮筋汤治疗KOA的关键靶点[促分裂原活化的蛋白激酶(mitogen activated protein kinase,MAPK)1、Akt激酶(akt kinase,AKT)1、淀粉样前体蛋白、Harvey大鼠肉瘤病毒致癌基因同源物、MAPK8、白细胞介素(interleukin,IL)-6、激肽原1、血管内皮生长因子A、热激蛋白90AA1、JUN原癌基因、肿瘤坏死因子(tumor necrosis factor,TNF)、MAPK14、IL-2、表皮生长因子受体、凝血因子Ⅱ],5个补肾壮筋汤治疗KOA的主要活性成分(汉黄芩素、啤酒甾醇、槲皮素、黄芩素、山奈酚)。GO富集分析结果显示,补肾壮筋汤治疗KOA主要涉及RNA聚合酶Ⅱ启动子转录的调控、凋亡过程的负调控、信号传导、调控细胞增殖、转录等生物过程,蛋白质结合、ATP结合、锌离子结合、相同蛋白质结合、酶结合等分子功能,以及细胞内质膜、细胞质、细胞核、胞质溶胶、核质等细胞组分; KEGG富集分析共筛选出磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)-AKT、MAPK、TNF等3条主要信号通路。分子对接结果显示,5个主要活性成分与15个关键靶点均有良好的结合能力,结合能均≤-5.0 kJ·mol-1; 分子对接图显示,活性成分大多嵌入靶点活性口袋的疏水区,结合模式多以π-π作用和氢键为主,提示其具有较好的结合活性,结合后构象稳定。结论:补肾壮筋汤治疗KOA的作用机制可能是通过汉黄芩素、啤酒甾醇、槲皮素、黄芩素、山奈酚等活性成分,参与调控RNA聚合酶Ⅱ启动子转录、凋亡过程、信号传导、细胞增殖、转录,影响PI3K-AKT、MAPK、TNF等信号通路。
Abstract:
Objective:To explore the mechanism of Bushen Zhuangjin Tang(补肾壮筋汤,BSZJT)in treatment of knee osteoarthritis(KOA)by using network pharmacological approach.Methods:The active ingredients of BSZJT were screened via retrieving traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP)using the bioinformatics analysis tool for molecular mechanism of traditional Chinese medicine(BATMAN-TCM),and their corresponding targets were screened out from PubChem database and Swiss TargetPrediction database for predicting the action targets of BSZJT.The action targets of BSZJT against KOA were obtained through overlapping the targets of BSZJT with KOA that retrieved from the GeneCard database,and they were analyzed by using String database,furthermore,the target protein-protein interaction(PPI)network of BSZJT against KOA was built by using Cytoscape3.7.2 software for screening the key targets of BSZJT against KOA.The active ingredients related to the action targets of BSZJT against KOA were screened out and were input into the Cytoscape3.7.2 software together with the action targets of BSZJT against KOA and the Chinese materia medica(CMM)in BSZJT,and the herb-active ingredient-target network of BSZJT against KOA was constructed for analysing the main active ingredients of BSZJT against KOA.The Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were performed on the target genes of BSZJT against KOA by using DAVID database and R3.6.2 software to dig biological functions and pathways of the target genes.Moreover,the molecular docking was conducted between the main active ingredients of BSZJT against KOA and its key targets by using Autodock vina software,and their binding energy was evaluated with the results visualized by Pymol software.Results:Ninety-nine active ingredients of BSZJT were screened out,and 768 corresponding targets were predicted,meanwhile,172 targets of BSZJT against KOA were obtained through overlapping the 989 KOA targets with the 768 predicted BSZJT targets.As revealed by the analysis,15 key targets(mitogen activated protein kinase(MAPK)1,akt kinase(AKT)1,amyloid precursor protein(APP),harvey rat sarcoma virus oncogene homolog(HRAS),MAPK8,interleukin(IL)-6,kininogen 1(KNG1),vascular endothelial growth factor A(VEGFA),heat shock protein 90AA1(HSP90AA1),JUN proto-oncogene(JUN),tumor necrosis factor(TNF),MAPK14,IL-2,epidermal growth factor receptor(EGFR)and blood coagulation factorⅡ(F2))and 5 main active ingredients(wogonin,cerevisterol,quercetin,baicalein and kaempferol)of BSZJT against KOA were obtained.The results of GO function enrichment analysis indicated that the target genes of BSZJT against KOA mainly participated in such biological processes(BPs)as regulation of transcription from RNA polymeraseⅡpromoter,negative regulation of apoptotic process,signal transduction,regulation of cell proliferation and transcription,the molecular functions(MFs)as protein binding,ATP binding,zinc ion binding,identical protein binding,enzyme binding,and the cellular components(CCs)as intracellular plasma membrane,cytoplasm,nucleus,cytosol,nucleoplasm.The results of KEGG pathway enrichment analysis showed that the target genes of BSZJT against KOA mainly acted on 3 major signaling pathways,such as signaling pathways of phosphatidylinositol 3-kinase(PI3K)-AKT,MAPK and TNF.The results of molecular docking showed that the 5 main active ingredients had good binding ability to the 15 key targets with the binding energy≤-5.0 kJ/mol.The molecular docking diagram illustrated that the active ingredients were mostly embedded in the hydrophobic region of the active pocket in targets with the binding modes mainly as π-π interaction and hydrogen bonds,which suggested they had good binding activity and the conformation was stabilized after binding.Conclusion:The mechanism of BSZJT against KOA may be that it participate in the BPs including regulation of transcription from RNA polymeraseⅡpromoter,apoptotic process,signal transduction,cell proliferation and transcription to affect the signaling pathways of PI3K-AKT,MAPK and TNF through the active ingredients of wogonin,cerevisterol,quercetin,baicalein and kaempferol.

参考文献/References:

[1] 中国中医药研究促进会骨伤科分会.膝骨关节炎中医诊疗指南(2020年版)[J].中医正骨,2020,32(10):1-14.
[2] LIPINSKI C A.Rule of five in 2015 and beyond:target and ligand structural limitations,ligand chemistry structure and drug discovery project decisions[J].Adv Drug Deliv Rev,2016,101:34-41.
[3] ABUSARAH J,BENABDOUNE H,SHI Q,et al.Elucidating the role of protandim and 6-gingerol in protection against osteoarthritis[J].J Cell Biochem,2017,118(5):1003-1013.
[4] CHOWDHURY N S,SOHRAB M H,RANA M S,et al.Cytotoxic naphthoquinone and azaanthraquinone derivatives from an endophytic fusarium solani[J].J Nat Prod,2017,80(4):1173-1177.
[5] LEYVA-LPEZ N,GUTIERREZ-GRIJALVA E P,AMBRIZ-PEREZ D L,et al.Flavonoids as cytokine modulators:a possible therapy for inflammation-related diseases[J].Int J Mol Sci,2016,17(6):921.
[6] GUIMARÃES G C,PIVA H,ARAUJO G C,et al.Binding investigation between M2-1protein from hRSV and acetylated quercetin derivatives:1H NMR,fluorescence spectroscopy,and molecular docking[J].Int J Biol Macromol,2018,111:33-38.
[7] KIM J R,YOO J J,KIM H A.Therapeutics in osteoarthritis based on an understanding of its molecular pathogenesis[J].Int J Mol Sci,2018,19(3):674.
[8] ZHANG X H,XU X X,XU T.Ginsenoside ro suppresses interleukin-1β-induced apoptosis and inflammation in rat chondrocytes by inhibiting NF-κB[J].Chin J Nat Med,2015,13(4):283-289.
[9] 罗晓平,于之伦,邓超,等.黄芩素对2,4,6-三硝基苯磺酸诱导的小鼠实验性肠炎的作用和机制[J].中国药理学与毒理学杂志,2017,31(6):541-546.
[10] 杨波,龙慧,王郑钢,等.黄芩素通过促进miR-29a-3p抑制MAP2K6改善骨性关节炎模型大鼠软骨细胞退化[J].中药药理与临床,2019,35(6):32-37.
[11] IMRAN M,RAUF A,SHAH Z A,et al.Chemo-preventive and therapeutic effect of the dietary flavonoid kaempferol:a comprehensive review[J].Phytother Res,2019,33(2):263-275.
[12] KADIOGLU O,NASS J,SAEED M E,et al.Kaempferol is an anti-inflammatory compound with activity towards NF-κB pathway proteins[J].Anticancer Res,2015,35(5):2645-2650.
[13] LI Q,WU M,FANG G,et al.MicroRNA-186-5p downregulation inhibits osteoarthritis development by targeting MAPK1[J].Mol Med Rep,2021,23(4):253.
[14] YANG Y,GU Y,ZHAO H,et al.Loganin attenuates osteoarthritis in rats by inhibiting IL-1β-induced catabolism and apoptosis in chondrocytes via regulation of phosphatidylinositol 3-kinases(PI3K)/Akt[J].Med Sci Monit,2019,25:4159-4168.
[15] WU X Y,LIU W T,WU Z F,et al.Identification of HRAS as cancer-promoting gene in gastric carcinoma cell aggressiveness[J].Am J Cancer Res,2016,6(9):1935-1948.
[16] CHEN Y T,HUANG Z Y,TANG H H,et al.Pterostilbene sensitizes cisplatin-resistant human bladder cancer cells with oncogenic HRAS[J].Cancers(Basel),2020,12(10):2869.
[17] LÜ G,LI L,WANG B,et al.LINC00623/miR-101/HRAS axis modulates IL-1β-mediated ECM degradation,apoptosis and senescence of osteoarthritis chondrocytes[J].Aging(Albany NY),2020,12(4):3218-3237.
[18] JIANG H,DAI J,ZHANG C,et al.Circ_0045714 alleviates TNF-α-induced chondrocyte injury and extracellular matrix degradation through miR-218-5p/HRAS axis[J].J Bioenerg Biomembr,2021,53(1):97-107.
[19] YUDUSHKIN I.Control of Akt activity and substrate phosphorylation in cells[J].IUBMB Life,2020,72(6):1115-1125.
[20] 颜春鲁,李盛华,安方玉,等.右归丸通过PI3K/Akt/mTOR信号通路对膝骨关节炎模型鼠软骨组织保护作用的研究[J].中国骨质疏松杂志,2020,26(3):318-322.
[21] CHU X, YU T, HUANG X,et al.Tomatidine suppresses inflammation in primary articular chondrocytes and attenuates cartilage degradation in osteoarthritic rats[J].Aging(Albany NY),2020,12(13):12799-12811.
[22] 谢文鹏,杨灵森,马亮,等.基于p38 MAPK信号通路探讨苍膝通痹胶囊对KOA大鼠关节软骨的保护作用[J].中国实验方剂学杂志,2020,26(18):70-76.

相似文献/References:

[1]樊庆阳,任凯晶.定制3D打印切模辅助全膝关节置换术治疗 膝骨关节炎合并股骨干骨折畸形愈合[J].中医正骨,2015,27(11):37.
[2]刘晓雅,孙永强,刘国杰.主动快速康复锻炼对全膝关节置换术后关节活动度的影响[J].中医正骨,2015,27(09):73.
[3]郑春松,叶蕻芝,李西海,等.透骨消痛胶囊中补肾柔肝药和活血祛风药治疗 骨关节炎作用方式的计算机模拟比较[J].中医正骨,2015,27(07):6.
　ZHENG Chunsong,YE Hongzhi,LI Xihai,et al.Comparison of the mode of action of Bushen Rougan(补肾柔肝)drugs versus Huoxue Qufeng(活血祛风)drugs contained in Tougu Xiaotong Jiaonang(透骨消痛胶囊)for the treatment of osteoarthritis:A computer simulation study[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(07):6.
[4]宋兵华,孙俊英,倪增良,等.全膝关节置换术前CT测量股骨后髁角的临床意义[J].中医正骨,2015,27(07):38.
[5]郑春松,叶蕻芝,李西海,等.独活寄生汤含药血清对白细胞介素1β诱导的 退变关节软骨细胞中基质金属蛋白酶 和环氧化酶2表达的影响[J].中医正骨,2015,27(12):1.
　ZHENG Chunsong,YE Hongzhi,LI Xihai,et al.Impact of Duhuo Jisheng Tang(独活寄生汤)medicated serum on expression of matrix metalloproteinase and cyclooxygenase 2 in degenerative articular chondrocytes induced by interleukin-1 beta[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(07):1.
[6]王金良,孙京涛,李玲,等.骨水泥联合螺钉修复全膝关节置换术中 胫骨平台内侧骨缺损[J].中医正骨,2015,27(12):55.
[7]冯荣,王平,李炳奇,等.铍针刺络拔罐结合中药口服治疗膝骨关节炎合并 原发性血小板增多症1例[J].中医正骨,2015,27(12):73.
[8]蔡云仙.围手术期耳穴按压联合平衡针疗法 在全膝关节置换术后镇痛中的应用[J].中医正骨,2015,27(06):41.
[9]张荣,王健.人工全膝关节置换术的围手术期心理护理[J].中医正骨,2015,27(05):77.
[10]喻长纯,杨明路,王战朝.不同手术方式治疗胫骨平台骨折畸形愈合的体会[J].中医正骨,2015,27(03):37.
[11]孟维娜,明立功,王新德,等.关节镜下清理联合腓骨近1/3段截骨治疗膝骨关节炎[J].中医正骨,2015,27(11):40.
[12]明立功,孟维娜,王新德,等.腓骨近端截骨治疗内侧间室膝骨关节炎的近期疗效观察[J].中医正骨,2015,27(10):25.
[13]张杰,王人彦,张玉柱.膝骨关节炎的治疗进展[J].中医正骨,2015,27(10):68.
[14]梁朝,蔡静怡,闫立,等.针刀疗法改善膝骨关节炎早期疼痛症状的疗效评价[J].中医正骨,2015,27(09):9.
　LIANG Zhao,CAI Jingyi,YAN Li,et al.Evaluation of the curative effect of needle-knife therapy for relieving knee pain in patients with early knee osteoarthritis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(07):9.
[15]王建武,党建军,李强,等.四联疗法治疗膝骨关节炎[J].中医正骨,2015,27(08):44.
[16]刘红娟,郭会利,郭树农.云克联合中药治疗膝骨关节炎的护理[J].中医正骨,2015,27(08):75.
[17]陈卫衡.探索建立系统的膝骨关节炎中医临床科研范式 和理论体系[J].中医正骨,2015,27(07):1.
[18]帅波,沈霖,杨艳萍,等.加味青娥丸治疗膝骨关节炎的作用机制研究[J].中医正骨,2015,27(07):15.
　SHUAI Bo,SHEN Lin,YANG Yanping,et al.Study on the mechanism of action of Jiawei Qing'e Wan(加味青娥丸)for the treatment of knee osteoarthritis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(07):15.
[19]梅其杰,袁长深,段戡,等.壮药骨痹方烫熨联合运动疗法治疗膝骨关节炎的临床研究[J].中医正骨,2015,27(07):27.
　MEI Qijie,YUAN Changshen,DUAN Kan,et al.Clinical study of the curative effect of hot compressing and rubbing with packet of Gubi Fang(骨痹方)combined with exercise therapy in the treatment of knee osteoarthritis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(07):27.
[20]王丹辉,张燕,刘丽娟,等.重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白 关节腔注射联合中药薰洗治疗膝骨关节炎的临床研究[J].中医正骨,2015,27(07):31.
　WANG Danhui,ZHANG Yan,LIU Lijuan,et al.Clinical study on intra-articular injection of TypeⅡrecombinant human tumor necrosis factor receptor-Fc fusion protein combined with Chinese herbal steaming and washing therapy for treatment of knee osteoarthritis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(07):31.

备注/Memo

备注/Memo:
基金项目:广西中医药适宜技术开发与推广项目(GZSY21-78) 通讯作者:欧志学 E-mail:ouzhixue1028@126.com
更新日期/Last Update: 1900-01-01