基于网络药理学探讨地榆抗菌活性成分研究及初步实验验证  

Study on the Antibacterial active ingredients of Sanguisorba officinalis based on network pharmacology and preliminary experimental verification

作　　者：朱洁[1] 张斌[1] 王英英 陈勇[1] 褚以文[1] 倪彩霞[1] Zhu Jie;Zhang Bin;Wang Ying-ying;Chen Yong;Chu Yi-wen;Ni Cai-xia(Antibiotics Research and Re-evaluation Key Laboratory of Sichuan Province,Sichuan Industrial Institute of Antibiotics,School of Pharmacy,Chengdu University,Chengdu 610106)

机构地区：[1]抗生素研究与再评价四川省重点实验室,四川抗菌素工业研究所,药学院,成都大学,成都610106

出　　处：《中国抗生素杂志》2022年第11期1162-1171,共10页Chinese Journal of Antibiotics

基　　金：国家重大新药创制项目(No.2019ZX09721001-006-002)。

摘　　要：目的采用网络药理学方法探讨地榆抗菌活性成分,并结合抗菌药效学实验进行验证。方法基于TCMSP数据库获取地榆化学成分及其对应靶点,通过Gene cards和OMIM数据库获取细菌感染靶点。通过Cytoscape构建药物-疾病-靶点网络图,利用STRING数据库构建蛋白互作网络,采用DAVID数据库开展预测靶点的GO和KEGG分析。采用琼脂二倍稀释法对预测的活性成分进行体外最低抑菌浓度(MIC)测定,进一步利用临床分离菌耐甲氧西林金黄色葡萄球菌(MRSA)和铜绿假单胞菌(Pseudomonas aeruginosa,PA)建立秀丽隐杆线虫感染和药物救治模型,观察线虫的存活情况,计算存活率。结果共筛选出8个潜在活性成分,其中山奈酚和槲皮素Degree值高于平均Degree值,得到17个关键靶点;GO和KEGG富集分析显示地榆抗细菌感染主要涉及细胞增殖、细胞凋亡、免疫应答等生物学过程,主要作用与调控Toll样受体、TNF、MAPK等信号通路相关。山奈酚对革兰阳性菌MRSA和MSSA的MIC_(50)和MIC_(90)均为128μg/mL。山奈酚、槲皮素对革兰阴性菌铜绿假单胞菌的MIC_(50)和MIC_(90)均为128μg/mL。与菌对照组比较,山奈酚(2.56 mg/mL)对MRSA和铜绿假单胞菌感染的秀丽隐杆线虫存活率明显升高(P<0.05),槲皮素(5.12 mg/mL)可明显提高铜绿假单胞菌感染线虫的存活率(P<0.05)。结论该研究初步揭示了山奈酚和槲皮素可能是地榆抗菌作用的物质基础之一,其作用机制可能与网络药理学预测的调节机体免疫和炎症反应等有关,为进一步实验研究奠定一定基础。Objective Based on the results of preliminary research in the laboratory,network pharmacology methods were used to explore the antibacterial active components and verified it with antibacterial pharmacodynamic experiments.Methods Based on the TCMSP database to obtain the chemical constituents and corresponding targets of Sanguisorba officinalis,obtain bacterial infection targets through Gene cards and OMIM database,and map the drugs and disease targets to obtain the predicted targets of the chemical composition of Sanguisorba officinalis.The Cytoscape software was used to construct a component-target-disease network diagram,STRING database was used to build a protein interaction network,GO and KEGG analysis was constructed by DAVID database.The agar double dilution method was used to determine the minimum inhibitory concentration(MIC)of the predicted active ingredients.The clinical isolates of methicillin-resistant Staphylococcus aureus(MRSA)and Pseudomonas aeruginosa(PA)were used to infect Caenorhabditis elegans(C.elegans).Established infection and drug treatment models under solid conditions,and verified the antibacterial activity of the predicted ingredients by observing the survivals of Caenorhabditis elegans.Results Sanguisorba officinalis could have eight potential active ingredients,the degree values of kaempferol and quercetin were higher than the average degree value.There were 88 targets that may be related to the anti-drug resistant bacteria,while 17 key targets were obtained;GO and KEGG enrichment analysis showed that the resistance to bacterial infection of Sanguisorba officinalis was mainly involved in biological processes such as cell proliferation,cell apoptosis,and immune response,and the main effects were related to the regulation of Toll-like receptor,TNF,MAPK,and other signaling pathways.The results of MIC determination showed that the MIC_(50) and MIC_(90) of kaempferol against Gram-positive bacteria MRSA and MSSA were both 128μg/mL.The MIC_(50) and MIC_(90) of kaempferol and quercetin again

关 键 词：地榆 抗菌活性成分 网络药理学 初步实验验证 

分 类 号：R978.1[医药卫生—药品]