摘要
为探讨天麻有效部位抗炎物质基础和作用机制,以天麻为原料,采用不同极性溶剂进行逐级提取,分别得到石油醚提取物、二氯甲烷提取物、乙酸乙酯提取物、正丁醇提取物和水提取物,并测定了各提取物的活性成分及生物活性,以此筛选天麻有效部位。通过超高效液相色谱-四极杆飞行时间质谱(ultra-high performance liquid chromatography-quadrupole-time of flight-mass spectrometry,UPLC-Q-TOF-MS/MS)技术对天麻有效部位进行成分鉴定,并利用网络药理学预测天麻有效部位潜在抗炎活性成分及作用靶点,将潜在活性成分与治疗炎症的核心靶点进行分子对接。结果表明:乙酸乙酯提取物中具有较高含量的活性成分,总多酚、总黄酮和多糖的含量分别为(164.69±1.57)、(64.81±3.21)mg/g和(279.64±3.11)mg/g;各提取物均具有较强的体外抗氧化和抗炎活性,其中乙酸乙酯提取物展现出最强的1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除能力和体外抗炎活性。相关性分析显示,总多酚含量与DPPH自由基清除能力半抑制浓度(half maximal inhibitory concentration,IC_(50))和2,2’-联氮双(3-乙基苯并噻唑啉-6-磺酸)阳离子自由基清除能力IC_(50)呈显著相关(P<0.05)和极显著相关(P<0.01),相关系数分别-0.587和-0.873;总黄酮含量分别与NO抑制能力IC_(50)和牛血清白蛋白变性抑制能力IC_(50)呈显著相关(P<0.05)和极显著相关(P<0.01),相关系数分别-0.780和-0.842。同时,采用UPLC-Q-TOF-MS/MS技术从天麻乙酸乙酯提取物中共鉴定出164个化学成分,网络药理学获得核心活性成分为5-(5-Methoxycarbonyl-5,8a-dimethyl-2-methylidene-3,4,4a,6,7,8-hexahydro-1H-naphthalen-1-yl)-3-methylpentanoic acid、4,4,8,10,14-Pentamethyl-17-(4,5,6-trihydroxy-6-methylheptan-2-yl)-2,5,6,7,9,15-hexahydro-1H-cyclopenta[a]phenanthrene-3,16-dione、Simvastatin等,关键靶点包括TP53、SRC、PIK3R1、PIK3CA、AKT1等,主要涉及到癌症有关通路、丝裂原活化蛋白激酶、环磷酸腺苷等信号通路,分子对接结果显示核心活性成分与关键靶点具有较好的结合活性。
This study was carried out in order to explore the anti-inflammatory constituents of the tuber of Gastrodia elata B1 and their mechanisms of action.The petroleum ether extract and its sequential fractions:dichloromethane,ethyl acetate,n-butanol and water-soluble fractions were determined for their active ingredients as well as assessed for their bioactivities.The bioactive ingredients were identified by ultra-high performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF-MS/MS),and network pharmacology was utilized to predict the potential anti-inflammatory components of the selected fraction and their targets.Furthermore,molecular docking was used to investigate the binding of the potential anti-inflammatory components to the core targets for treating inflammation.The results showed that the ethyl acetate fraction possessed the highest content of bioactive ingredients,with total polyphenol,total flavonoid and polysaccharide contents of(164.69±1.57),(64.81±3.21)and(279.64±3.11)mg/g,respectively.The ethanol extract and its fractions possessed strong in vitro antioxidant and anti-inflammatory activities,among which the ethyl acetate fraction demonstrated the strongest 1,1-diphenyl-2-picrylhydrazyl(DPPH)scavenging capacity and in vitro anti-inflammatory activity.Correlation analysis showed that the total polyphenol content was significantly(P<0.05)and highly significantly(P<0.01)correlated with the half maximal inhibitory concentration(IC50)against DPPH radical and 2,2’-biazobis(3-ethylbenzothiazoline-6-sulfonic acid)cationic radical,with correlation coefficients of-0.587 and-0.873,respectively.The total flavonoid content was significantly(P<0.05)and highly significantly(P<0.01)correlated with the IC50 values against NO and the denaturation of bovine serum albumin(BSA),with correlation coefficients of-0.780 and-0.842,respectively.Meanwhile,a total of 164 chemical constituents were identified from the ethyl acetate fraction,and core active ingredients including 5-(5-methoxycarbonyl-5,8a-dimethyl-2-methylidene-3,4,4a,6,7,8-hexahydro-1H-naphthalen-1-yl)-3-methylpentanoic acid,4,4,8,10,14-pentamethyl-17-(4,5,6-trihydroxy-6-methylheptan-2-yl)-2,5,6,7,9,15-hexahydro-1H-cyclopenta[a]phenanthrene-3,16-dione,and simvastatin and key targets including TP53,SRC,PIK3R1,PIK3CA,and AKT1,which were mainly involved in cancer-related signaling pathways as well as the mitogen-activated protein kinase(MAPK)and cyclic adenosine monophosphate(cAMP)signaling pathways,were obtained by network pharmacology.The results of molecular docking showed that the core active ingredients had good binding activity with the key targets.
作者
徐柠檬
董仕豪
秦粉
申开泽
李为兰
华燕
郭磊
XU Ningmeng;DONG Shihao;QIN Fen;SHEN Kaize;LI Weilan;HUA Yan;GUO Lei(College of Biology and Food Engineering,Southwest Forestry University,Kunming 650224,China;Yunnan Key Laboratory of Gastrodia and Fungi Symbiotic Biology,Zhaotong University,Zhaotong 657000,China;Yunnan Engineering Research Center of Green Planting and Processing of Gastrodia,Zhaotong University,Zhaotong 657000,China)
出处
《食品科学》
北大核心
2025年第4期30-43,共14页
Food Science
基金
云南省天麻与真菌共生生物学重点实验室开放课题(TMKF2024B05)
云南省科技厅农业联合专项(202101BD070001-054)
云南省科技厅基础研究计划项目(202101AT070042)。
关键词
天麻
活性成分
抗氧化活性
抗炎活性
网络药理学
分子对接
Gastrodia elata B1
active constituents
antioxidant activity
anti-inflammatory activity
network pharmacology
molecular docking
作者简介
第一作者:徐柠檬(1998-)(ORCID:0009-0002-6585-5811),男,硕士研究生,研究方向为天然产物活性成分开发与利用。E-mail:xuningmeng1007@163.com;通信作者:申开泽(1987-)(ORCID:0000-0002-0441-4257),男,讲师,硕士,研究方向为高原特色生物资源开发与利用。E-mail:ztushenkz@163.com;通信作者:郭磊(1981-)(ORCID:0000-0003-3587-742X),男,副教授,博士,研究方向为天然产物活性成分开发与利用。E-mail:guoleigift.student@sina.com。
