基于网络药理学的鹿茸治疗骨关节炎作用的机制  

Mechanism of Cervi Cornu Pantotrichum in Treatment of Osteoarthritis Based on Network Pharmacology

作　　者：郭子奕 杨扬[1] 何忠梅[1] 时坤[1] 李健明[1] 陈维佳 宗颖[1] 杜锐[1] GUO Zi-yi;YANG Yang;HE Zhong-mei;SHI Kun;LI Jian-ming;CHEN Wei-jia;ZONG Ying;DU Rui(Jilin Agricultural University,Changchun 130000,China)

机构地区：[1]吉林农业大学,长春130000

出　　处：《中国实验方剂学杂志》2022年第4期194-204,共11页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家重点研发计划项目(2018YFC1706603-02);吉林省科技发展计划项目(20200101006FG);吉林省重点技术研发项目(20180201010YY-1)。

摘　　要：目的:利用网络药理学方法探索鹿茸(Cervi Cornu Pantotrichum)在治疗骨关节炎中的作用机制。方法:利用BATMAN-TCM数据库筛选出鹿茸的有效活性成分,挖掘对应的靶点信息。通过GeneCards,在线人类孟德尔遗传数据库(OMIM)得到骨关节炎相关靶点信息。将成分靶点和骨关节炎靶点取交集,使用STRING平台建立蛋白质-蛋白质相互作用(PPI)网络。使用DAVID数据库对鹿茸治疗骨关节炎作用靶点进行基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)通路分析,利用R x643.6.3软件绘制GO,KEGG富集分析高级气泡图,利用Cytoscape 3.7.2软件绘制"中药-成分-靶点-通路"网络。体外实验分别检测鹿茸对产生氧化损伤和炎症的RAW 264.7细胞的细胞活力和肿瘤坏死因子(TNF)-α水平。结果:共计得到鹿茸活性成分20种,其中神经酰胺和6’-O-β-D-葡糖基龙胆苦苷、脑苷、橄榄苦苷、鞘磷脂、胆固醇阿魏酸酯没有达到筛选条件的得分,所以共筛选出14种有效活性成分。共计303个有效成分靶点,共得到3093个骨关节炎作用靶点。将成分靶点与骨关节炎作用靶点取交集,得到活性成分-骨关节炎作用靶点92个。GO富集分析和KEGG通路分析显示,鹿茸涉及的生物过程主要有氧化还原过程,RNA聚合酶Ⅱ启动子转录的正调控、炎症反应、蛋白质合成、破骨细胞分化,TNF信号通路、癌症通路、哺乳动物雷帕霉素靶蛋白(mTOR)信号通路、丝裂原活化蛋白激酶(MAPK)信号通路、环磷酸腺苷(cAMP)信号通路等生物过程和信号通路。体外实验结果显示,一定浓度的鹿茸蛋白显著增加了过氧化氢(H2O2)诱导的氧化损伤RAW 264.7细胞的细胞活力(P<0.05,P<0.01),并且降低了脂多糖(LPS)诱导炎症RAW 264.7细胞的TNF-α水平(P<0.05)。结论:基于网络药理学方法阐释了鹿茸多成分、多靶点、多途径的治疗OA作用机制,证实了鹿茸具有抗炎及抗氧化活性,为深入研究�Objective:To explore the mechanism of Cervi Cornu Pantotrichumin in the treatment of osteoarthritis by network pharmacology.Method:The active ingredients and the corresponding targets of Cervi Cornu Pantotrichumin were screened out by a Bioinformatics Analysis Tool of Molecular mechANism of Traditional Chinese Medicine(BATMAN-TCM).The targets related to osteoarthritis were obtained through GeneCards and Online Mendelian Inheritance in Man(OMIM).The targets corresponding to the active ingredients and those related to osteoarthritis were intersected to reveal the common targets,and STRING was adopted to build a protein-protein interaction(PPI)network.DAVID was used for gene ontology(GO)annotation and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment on the antiosteoarthritis targets of Cervi Cornu Pantotrichumin,and R x643.6.3 was employed to produce the advanced bubble charts of GO terms and KEGG pathways.Cytoscape 3.7.2 was used to establish the“Chinese medicinal herb-active ingredient-target-signaling pathway”network.In vitro experiments were performed to detect the viability of RAW 264.7 cells exposed to oxidative stress and the tumor necrosis factor(TNF)-αlevel in RAW264.7 cells with inflammation under the treatment by Cervi Cornu Pantotrichumin.Result:A total of 20 active ingredients of Cervi Cornu Pantotrichum were obtained,of which ceramide,6’-O-β-D-glucosylgentiopicroside,cerebroside,oleuropein,sphingomyelin,and cholesterol ferulate did not meet the screening conditions.Therefore,a total of 14 active ingredients were finally screened out,and 303 and 3093 targets of active ingredients and osteoarthritis were respectively obtained.The two target sets were taken to intersect,which revealed 92 common targets.GO annotation and KEGG pathway enrichment showed that the targets were mainly involved in redox process,positive regulation of RNA polymeraseⅡpromoter transcription,inflammatory response,protein synthesis,osteoclast differentiation,TNF signaling pathway,signaling pathways in cancer,

关 键 词：鹿茸 骨关节炎 网络药理学 作用机制 信号通路 氧化应激 炎症 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学]