基于网络药理学和分子对接分析赤芍抗氧化应激作用的活性成分及其机制

doi:10.16431/j.cnki.1671-7236.2023.03.037

摘要/Abstract

摘要： 【目的】利用网络药理学及分子对接技术分析赤芍抗氧化应激作用的有效活性成分及其分子机制。【方法】运用TCMSP数据库搜集赤芍的活性成分和靶点,借助UniProt数据库对靶点进行基因名转化;通过GeneCards和OMIM数据库检索氧化应激的相关基因,利用Veeny在线平台获得赤芍与氧化应激的交集靶点,将交集靶点上传至STRING数据库和Cytoscape 3.8.0软件建立蛋白互作(PPI)网络图和赤芍-靶点-氧化应激可视化网络并进行拓扑学分析,筛选关键靶点;运用Metascape数据库对关键靶点进行GO功能和KEGG通路富集分析,确定赤芍发挥抗氧化应激作用的活性成分和信号通路;运用AutoDock和PyMol软件对赤芍发挥抗氧化应激作用的核心靶点进行分子对接验证。【结果】共筛选到12个赤芍活性成分,包括黄芩素、β-谷甾醇、鞣花酸、豆甾醇、(+)-儿茶素等;赤芍活性成分与氧化应激靶点分别有76和4 873个,其中赤芍与氧化应激交集靶点共69个,包括蛋白激酶B(AKT1)、转录因子AP-1(JUN)、细胞肿瘤抗原p53(TP53)、肿瘤坏死因子(TNF)和半胱氨酸蛋白酶-3(CASP3)等;GO功能和KEGG通路富集结果提示,赤芍抗氧化应激与磷脂酰肌醇-3-激酶/蛋白激酶B(PI3K-Akt)、核因子κB(NF-κB)等多个信号通路有关;分子对接结果表明,赤芍主要活性成分黄芩素、β-谷甾醇与核心靶点有较好的结合能力。【结论】赤芍可能通过黄芩素、β-谷甾醇、鞣花酸等活性成分调控PI3K-Akt、NF-κB、白细胞介素-17(IL17)等信号通路中的关键靶点来发挥抗氧化应激作用。

关键词: 赤芍; 抗氧化应激; 作用机制; 网络药理学; 分子对接

Abstract: 【Objective】 The purpose of this experiment was to analyze the effective active ingredients of Radix Paeoniae Rubra on antioxidative stress and its molecular mechanism using network pharmacology and molecular docking technology.【Method】 Active ingredients and related targets of Radix Paeoniae Rubra were collected from TCMSP database and then converted into gene names using UniProt database.Targets related to oxidative stress were collected from GeneCards and OMIM databases,and the intersection targets of Radix Paeoniae Rubra and oxidative stress were obtained by Veeny online platform.Subsequently,protein-protein interaction (PPI) network map and a Radix Paeoniae Rubra-target-oxidative stress visualization network for topological analysis were established by STRING database and Cytoscape 3.8.0 software,and then key targets were identified.The active ingredients and antioxidant signaling pathways of Radix Paeoniae Rubra were determined by GO function and KEGG pathway enrichment analysis with Metascape database.AutoDock and PyMol softwares were used to perform molecular docking verification of the core target of Radix Paeoniae Rubra.【Result】 12 active ingredients in Radix Paeoniae Rubra were screened,including baicalein,beta-sitosterol,ellagic acid,stigmasterol,(+)-catechin,etc.There were 76 active ingredients targets in Radix Paeoniae Rubra and 4 873 oxidative stress targets,including 69 intersection targets,such as protein kinase B (AKT1),activator protein 1 (JUN),cellular tumor antigen p53 (TP53),tumor necrosis factor (TNF) and caspase-3 (CASP3), etc.The results of GO function and KEGG pathway enrichment analysis revealed that antioxidative stress of Radix Paeoniae Rubra might be related to the phosphatidylinositol-3-kinase-threonine-protein kinase (PI3K-Akt),neurofibromin-κB (NF-κB),and other signaling pathways.Moreover,the results of molecular docking showed that the main active ingredients of Radix Paeoniae Rubra,baicalein and beta sitosterol,had good binding ability to the core targets.【Conclusion】 The active ingredients such as baicalein,beta sitosterol and ellagic acid in Radix Paeoniae Rubra might play an antioxidative stress role by regulating key targets in PI3K-Akt,NF-κB,interleukin-17 (IL17) and other signaling pathways.

Key words: Radix Paeoniae Rubra; antioxidative stress; mechanism of action; network pharmacology; molecular docking

中图分类号:

S853.74

豆佳红, 王梓颖, 杨娟, 周炜炜, 张同存, 何红鹏, 戴小枫, 李秀梅. 基于网络药理学和分子对接分析赤芍抗氧化应激作用的活性成分及其机制[J]. 中国畜牧兽医, 2023, 50(3): 1229-1240.

DOU Jiahong, WANG Ziying, YANG Juan, ZHOU Weiwei, ZHANG Tongcun, HE Hongpeng, DAI Xiaofeng, LI Xiumei. Analysis of the Active Ingredients and Mechanism of Radix Paeoniae Rubra Antioxidative Stress Based on Network Pharmacology and Molecular Docking[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1229-1240.

参考文献

[1] NYSKA A,KOHEN R.Oxidation of biological systems:Oxidative stress phenomena,antioxidants,redox reactions,and methods for their quantification[J].Toxicologic Pathology,2002,30(6):620-650.
[2] SIES H.Oxidative stress:A concept in redox biology and medicine[J].Society for Redox Biology and Medicine,2015,4:180-183.
[3] ARUOMA O I,GROOTVELD M,BAHORUN T.Free radicals in biology and medicine:From inflammation to biotechnology[J].Biofactors,2006,27(1-4):1-3.
[4] BERG H V D,FAULKS R,GRANADO H F,et al.The potential for the improvement of carotenoid levels in foods and the likely systemic effects[J].Agricultural and Food Chemistry,2000,80(7):880-912.
[5] 安丽凤,鲁光宝,刘海洲,等.藁本内酯对H₂O₂诱导的B16黑素瘤细胞氧化损伤的保护作用[J].现代生物医学进展,2017,17(11):2032-2036. AN L F,LU G B,LIU H Z,et al.Protective effect of Z-ligustilide on the oxidation damage of B16 melanoma cells induced by H₂O₂[J].Progress in Modern Biomedicine,2017,17(11):2032-2036.(in Chinese)
[6] 陈松,滕勇,陈宝炳.氧化应激与2型糖尿病患者发病程度的相关性探讨[J].中国卫生检验杂志,2017,27(23):3446-3448. CHEN S,TENG Y,CHEN B B.Correlation between oxidative stress and the incidence of type 2 diabetes mellitus[J].Chinese Health Laboratory Technology,2017,27(23):3446-3448.(in Chinese)
[7] 陈渝春,冯兰超.癌症患者抗氧化态与氧化应激的变化[J].四川医学,2008,29(7):844-846. CHEN Y C,FENG L C.Changes of antioxidant status and oxidative stress in cancer patients[J].Sichuan Medical Journal,2008,29(7):844-846.(in Chinese)
[8] 邓映,钟建桥.皮肤光损伤与氧化应激研究进展[J].临床皮肤科杂志,2017,46(9):671-673. DENG Y,ZHONG J Q.Advances in skin photodamage and oxidative stress[J].Journal of Clinical Dermatology,2017,46(9):671-673.(in Chinese)
[9] 樊春荔,吴嘉,汪俊军.心血管疾病氧化应激反应相关microRNAs的研究进展[J].临床检验杂志,2015,33(10):779-781. FAN C L,WU J,WANG J J.Research progress of oxidative stress related microRNAs in cardiovascular diseases[J].Chinese Journal of Clinical Laboratory Science,2015,33(10):779-781.(in Chinese)
[10] 毛文星,李冰,王志梅,等.氧化应激、血管内皮功能障碍与高血压[J].现代生物医学进展,2014,14(19):3770-3774. MAO W X,LI B,WANG Z M,et al.Oxidative stress and endothelial dysfunction in hypertension[J].Progress in Modern Biomedicine,2014,14(19):3770-3774.(in Chinese)
[11] 潘婕,刘增援,梁政,等.赤芍水提物与抗菌药联合对耐药大肠杆菌抑菌作用的研究[J].中国畜牧兽医,2015,42(11):3080-3086. PAN J,LIU Z Y,LIANG Z,et al.Study on the anti-bacterial effects of aqueous extract of Radix Paeoniae Rubra combined with antibiotics against drug-resistant E.coli[J].China Animal Husbandry & Veterinary Medicine,2015,42(11):3080-3086.(in Chinese)
[12] 吴玲芳,王子墨,赫柯芊,等.赤芍的化学成分和药理作用研究概况[J].中国实验方剂学杂志,2021,27(18):198-206. WU L F,WANG Z M,HE K Q,et al.Chemical constituents and pharmacological effects of Paeoniae Radix Rubra:A review [J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(18):198-206.(in Chinese)
[13] 马秀凤,马浩如,钱翠萍,等.赤芍治疗肺心病的临床观察和防治肺动脉高压的实验研究[J].中西医结合杂志,1988,11:660-662. MA X F,MA H R,QIAN C P,et al.Experimental and clinical studies on treatment of pulmonary hear disease and pulmonary hypertension with Paeonia lactiflora[J].Chinese Journal of Integrated Traditional and Western Medicine,1988,11:660-662.(in Chinese)
[14] 何丽娜,何素冰,江怡,等.赤芍总苷对大鼠皮层神经细胞钙超载损伤的保护作用[J].中药新药与临床药理,2001,3:212-215. HE L N,HE S B,JIANG Y,et al.Profective effect of total pcaony glycoside on calium-overloading injury of nerve cells in rat cortex[J].Traditional Chinese Drug Research and Clinical Pharmacology,2001,3:212-215.(in Chinese)
[15] 林彦君,章津铭,瞿燕,等.赤芍总苷对实验性大鼠胃溃疡模型的影响[J].中国实验方剂学杂志,2010,16(6):215-217. LIN Y J,ZHANG J M,QU Y,et al.The impact of total paeony glycoside on experimental gastric ulcer in rat model[J].Chinese Journal of Experimental Traditional Medical Formulae,2010,16(6):215-217.(in Chinese)
[16] 郭春燕.几种中药活性成分对H₂O₂诱导SH-SY5Y细胞氧化损伤的影响及其机制研究[D].石家庄:河北医科大学,2013. GUO C Y.The effect and mechanism of several active components of traditional Chinese medicine on oxidative damage of SH-SY5Y cells induced by H₂O₂[D].Shijiazhuang: Hebei Medical University,2013.(in Chinese)
[17] 许文平,王艳艳,孟笑玮,等.芍药苷对ApoE^-/-小鼠血脂及主动脉斑块的影响[J].天津中医药大学学报,2014,33(4):210-212. XU W P,WANG Y Y,MENG X W,et al.Effect of paeoniflorin on blood lipids and atherosclerotic plaque in mice with apolipoprotein E deficient[J].Journal of Tianjin University of Traditional Chinese Medicine,2014,33(4):210-212.(in Chinese)
[18] JIANG F J,ZHAO Y L,WANG J B,et al.Comparative pharmacokinetic study of paeoniflorin and albiflorin after oral administration of Radix Paeoniae Rubra in normal rats and the acute cholestasis hepatitis rats[J].Fitoterapia,2012,83(2):415-421.
[19] LUO C,WANG H,CHEN X,et al.Protection of H9c2 rat cardiomyoblasts against oxidative insults by total paeony glucosides from Radix Paeoniae Rubrae[J].Phytomedicine,2013,21(1):20-24.
[20] ZHENG Q,ZHAO Y,ZHOU G,et al.Paeoniflorin protects against ANIT-induced cholestasis by ameliorating oxidative stress in rats[J].Food and Chemical Toxicology,2013,8(58):242-248.
[21] PARK S Y,SEETHARAMAN R,KO M J,et al.Ethyl linoleate from garlic attenuates lipopolysaccharide-induced pro-inflammatory cytokine production by inducing heme oxygenase-1 in RAW264.7 cells[J].International Immunopharmacology,2014,19(2):253-261.
[22] 高书锋,雷平,李咏梅,等.黄芩素对22~63日龄湘黄肉鸡生长性能、免疫功能、抗氧化能力、血脂状态及肉品质的影响[J].动物营养学报,2022,34(1):215-227. GAO S F,LEI P,LI Y M,et al.Effects of baicalein on growth performance,immune function,antioxidant ability,blood lipid status and meat quality of Hunan Yellow chickens during 22 to 63 days of age[J].Chinese Journal of Animal Nutrition,2022,34(1):215-227.(in Chinese)
[23] 张冬阳,郭雪松.薏米中β谷甾醇的分离纯化及其抗氧化活性的研究[J].食品工业科技,2018,39(11):205-210. ZHANG D Y,GUO X S.Isolation,purification and antioxidant activity of β-sitosterol from Coix chinensis Tod.[J].Science and Technology of Food Industry,2018,39(11):205-210.(in Chinese)
[24] 崔珊珊,毕凯媛,吴杰,等.响应面法优化树莓鞣花酸提取工艺及其体外抗氧化活性[J].食品工业科技,2019,40(1):149-155. CUI S S,BI K Y,WU J,et al.Optimization of extraction technology and antioxidative activity of ellagic acid in raspberry by response surface methodology[J].Science and Technology of Food Industry,2019,40(1):149-155.(in Chinese)
[25] 饶夙,陈祥贵,刘振平,等.鞣花酸和石榴皮多酚提取物抗氧化活性的比较[J].食品工业科技,2012,33(12):111-113. RAO S,CHEN X G,LIU Z P,et al.Comparative study of antioxidant activities of ellagic acid and pomegranate peel extract[J].Science and Technology of Food Industry,2012,33(12):111-113.(in Chinese)
[26] 张正平.β-arrestin调控细胞氧化应激凋亡中ASK1信号通路的研究[D].南京:南京师范大学,2009. ZHANG Z P.Study on the ASK1 signaling pathway in the regulation of oxidative stress apoptosis by β-arrestin[D].Nanjing:Nanjing Normal University 2009.(in Chinese)
[27] KIM A H,KHURSIGARA G,SUN X,et al.AKT phosphorylates and negatively regulates apoptosis signal-regulating kinase1[J].Molecular and Cellular Biology,2001,21(3):893-901.
[28] 李芊芊.氧化应激诱导细胞凋亡通路的研究进展[J].世界最新医学信息文摘,2019,19(12):63-64. LI Q Q.Advances in oxidative stress-induced apoptosis pathway[J].World Latest Medicine Information, 2019,19(12):63-64.(in Chinese)
[29] SEN T,SEN N,NOORDHUIS M,et al.OGDHL is a modifier of AKT-dependent signaling and NF-κB function [J].PLoS One 2012,7(11):e48770.
[30] 窦彩霞,王倩,安建勇,等.Keap1-Nrf2/ARE信号通路及其在畜禽抗氧化中的研究进展[J].中国畜牧兽医,2019,46(9):2567-2574. DOU C X,WANG Q,AN J Y,et al.Keap1-Nrf2/ARE signaling pathway and its research progress in antioxidation of livestock and poultry[J].China Animal Husbandry & Veterinary Medicine,2019,46(9):2567-2574.(in Chinese)
[31] DANDAN H,WEI C,XIAOLONG R S,et al.Cytoprotective effect of chlorogenic acid against hydrogen peroxide-induced oxidative stress in MC3T3-E1 cells through PI3K/Akt-mediated Nrf2/HO-1 signaling pathway[J].Oncotarget,2017,8(9):14680-14692.
[32] ZHUANG Y,WU H,WANG X,et al.Resveratrol attenuates oxidative stress-induced intestinal barrier injury through PI3K/Akt-mediated Nrf2 signaling pathway[J].Oxidative Medicine and Cellular Longevity,2019,2019:7591840.
[33] FENG Y,YU Y H,WANG S T,et al.Chlorogenic acid protects D-galactose-induced liver and kidney injury via antioxidation and anti-inflammation effects in mice[J].Pharmaceutical Biology,2016,54(6):1027-1034.
[34] 王丽雪,解玉怀,杨维仁,等.植物多糖通过核因子κB及丝裂原活化蛋白激酶/核因子E2相关因子2信号通路发挥抗氧化作用的机制[J].动物营养学报,2018,30(10):3857-3863. WANG L X,XIE Y H,YANG W R,et al.Plant polysaccharides:Mechanism of anti-oxidant action through nuclear factor κB and mitogen-activated protein kinase/nuclear factor erythroid 2-related factor 2 signaling pathways [J].Chinese Journal of Animal Nutrition,2018,30(10):3857-3863.(in Chinese)
[35] GANG D W,JIAN L W,XIU Z F,et al.Protein adducts of malondialdehyde and 4-hydroxynonenal contribute to trichloroethene-mediated autoimmunity via activating Th17 cells:Dose- and time-response studies in female MRL^+/+ mice[J].Toxicology,292(2-3):113-122.
[36] 李强,田杰,陈紫君,等.氧化应激上调IL-17水平在Graves眼病中的意义[J].第三军医大学学报,2011,33(10):1068-1070. LI Q,TIAN J,CHEN Z J,et al.Oxidative stress up-zegnlale IL-17 in patients with Graves’ ophthalmopathy[J]. Journal of Army Medical University,2011,33(10):1068-1070.(in Chinese)