广金钱草挥发油基于TRP通道的抗炎作用研究
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摘要
目的筛选广金钱草挥发油抗炎作用的关键化学成分及靶标蛋白。方法采用水蒸气蒸馏法提取广金钱草中的挥发油,通过气相色谱-质谱联用法鉴定其化学成分,采用峰面积归一化法测定其化学成分的相对含量。基于中药系统药理学技术平台(Traditional Chinese Medicine Systems Pharmacology,TCMSP)建立小分子配体库,借助Swiss Target Prediction在线进行反向靶标预测,通过KOBAX 3.0筛选抗炎通路,采用分子对接技术(SYBYL 2.1)将关键小分子与瞬时受体电位(TRP)通路中的靶标蛋白进行能量匹配,基于Cytoscape3.5.1构建化学成分-靶标网络模型。结果从广金钱草挥发油中共检测出48个色谱峰,通过质谱库及文献检索确定33种化合物结构,占挥发油总量的90.1%。关键化学成分17个,共筛选出88个靶标蛋白,TRP通路包括11个潜在靶标。分子对接发现叶绿醇、三十一烷、法尼基丙酮和角鲨烯等为广金钱草挥发油发挥抗炎作用的关键化学成分。TPRV1、PRKCB和PRKCD为抗炎关键靶标蛋白。结论初步筛选了广金钱草挥发油抗炎的关键靶点及活性成分,为其产品的开发和应用提供理论依据。
Objective To screen out the key chemical constituents and target protein of essential oil of Desmodium styracifolium for its anti-inflammatory effect. Methods Steam distillation method was used to extract the volatile oils from D. styraci folium, and its chemical constituents were identified by GC-MS, and the relative content of chemical constituents was determined by peak area normalization. The small molecule ligand library was established based on Traditional Chinese Medicine Systems Pharmacology(TCMSP). Reverse target prediction was conducted online using Swiss Target Prediction, the anti-inflammatory pathways were screened by KOBAX 3.0, conducting energy match between the key small molecular and the target protein in the TRP channels by molecular docking(SYBYL2.1). Construction of chemical constituents-targets network model was based on Cytoscape 3.5.1. Results A total of 48 chromatographic peaks were detected from D. styracifolium volatile oils, and 33 kinds of compound structure were determined by searching in mass spectral database and document retrieval, which account for 90.1% of total volatile oils. There were 17 key chemical constituents, and 88 target proteins were selected. TRP channels included 11 potential targets. Through molecular docking, we found that the phytol, hentriacontane, farnesyl acetone, and squalene were the key anti-inflammatory chemical constituents of D. styraci folium volatile oils. TPRV1(transient receptor potential cation channel, subfamily V, member 1), PRKCB(protein kinase C, beta), and PRKCD(protein kinase C, delta)(degree > 10) are the key anti-inflammatory target protein. Conclusion We preliminarily select the key anti-inflammatory target and active constituents of D. styraci folium volatile oils from this study, and this research provides the theoretical basis for the development and application of its products.
Objective To screen out the key chemical constituents and target protein of essential oil of Desmodium styracifolium for its anti-inflammatory effect. Methods Steam distillation method was used to extract the volatile oils from D. styraci folium, and its chemical constituents were identified by GC-MS, and the relative content of chemical constituents was determined by peak area normalization. The small molecule ligand library was established based on Traditional Chinese Medicine Systems Pharmacology(TCMSP). Reverse target prediction was conducted online using Swiss Target Prediction, the anti-inflammatory pathways were screened by KOBAX 3.0, conducting energy match between the key small molecular and the target protein in the TRP channels by molecular docking(SYBYL2.1). Construction of chemical constituents-targets network model was based on Cytoscape 3.5.1. Results A total of 48 chromatographic peaks were detected from D. styracifolium volatile oils, and 33 kinds of compound structure were determined by searching in mass spectral database and document retrieval, which account for 90.1% of total volatile oils. There were 17 key chemical constituents, and 88 target proteins were selected. TRP channels included 11 potential targets. Through molecular docking, we found that the phytol, hentriacontane, farnesyl acetone, and squalene were the key anti-inflammatory chemical constituents of D. styraci folium volatile oils. TPRV1(transient receptor potential cation channel, subfamily V, member 1), PRKCB(protein kinase C, beta), and PRKCD(protein kinase C, delta)(degree > 10) are the key anti-inflammatory target protein. Conclusion We preliminarily select the key anti-inflammatory target and active constituents of D. styraci folium volatile oils from this study, and this research provides the theoretical basis for the development and application of its products.
引文
[1]曾南,王建,夏厚林,等.芳香开窍药药理作用研究进展[J].中药药理与临床,2008,24(1):76-79.
[2]Campbell S T,Franks C E,Borne A L,et al.Chemoproteomic discovery of a ritanserin-targeted kinase network mediating apoptotic cell death of lung tumor cells[J].Mol Pharmacol,2018,94(5):1246-1255.
[3]程彬峰,侯媛媛,姜民,等.基于网络药理学的清肺消炎丸抗炎机制的初步研究[J].药学学报,2013,48(5):686-693.
[4]刘学,崔健,陈新.广金钱草现代研究进展[J].长春中医药大学学报,2006,22(4):84-85.
[5]刘茁,董焱,王宁,等.金钱草的化学成分[J].沈阳药科大学学报,2005,22(6):422-437.
[6]陈丰连,王术玲,徐鸿华.广金钱草挥发油的气相色谱-质谱分析[J].广州中医药大学学报,2015,22(4):302-303.
[7]刘瑞来,刘俊劭,林志銮,等.不同方法提取的金钱草叶挥发油化学成分的GC-MS分析[J].安徽农业科学,2012,40(12):7036-7037.
[8]Spitzer R,Jain A N.Surflex-Dock:Docking benchmarks and real-world application[J].J Comput Aided Mol Des,2012,26(6):687-699.
[9]Xie C,Mao X,Huang J,et al.KOBAS 2.0:A web server for annotation and identification of enriched pathways and diseases[J].Nucleic Acids Res,2011,doi:10.1093/nar/gkr483.
[10]Dainaa A,Michielin O,Zoete V.SwissADME:A free web tool to evaluate pharmacokinetics,drug-likeness and medicinal chemistry friendliness of small molecules[J].Sci Rep,2017,doi:10.1038/srep42717.
[11]林桂源,姚华聪,郑细娜,等.基于分子对接技术的常用降糖中药有效成分虚拟筛选[J].中国实验方剂学杂志,2015,21(15):202-206.
[12]Berman H M,Westbrook J,Feng Z,et al.The protein data bank[J].Nucleic Acids Res,2000,28(1):235-242.
[13]Lishko P V,Procko E,Jin X,et al.The ankyrin repeats of trpv1 bind multiple ligands and modulate channel sensitivity[J].Neuron,2007,54(6):905-918.
[14]Inada H,Procko E,Sotomayor M,et al.Structural and biochemical consequences of disease-causing mutations in the ankyrin repeat domain of the human TRPV4channel[J].Biochemistry,2012,51(31):6195-6206.
[15]Mc Cleverty C J,Koesema E,Patapoutian A,et al.Crystal structure of the human TRPV2 channel ankyrin repeat domain[J].Protein Sci,2006,15(9):2201-2206.
[16]Sutton R B,Sprang S R.Structure of the protein kinase Cbeta phospholipid-binding C2 domain complexed with Ca2+[J].Structure,1998,6(11):1395-1405.
[17]Shannon P,Markiel A,Ozier O,et al.Cytoscape:Asoftware environment for integrated models of biomolecular interaction networks[J].Genome Res,2003,13(11):2498-2504.
[18]Holzer P.Transient receptor potential(TRP)channels as drug targets for diseases of the digestive system[J].Pharmacol Ther,2011,131(1):142-170.
[19]Steinberg S F.Cardiac actions of protein kinase Cisoforms[J].Physiology,2012,27(3):130-139.
[20]Frazier W J,Xue J,Luce W A,et al.MAPK signaling drives inflammation in LPS-stimulated cardiomyocytes:The route of crosstalk to G-protein-coupled receptors[J].PLoS One,2012,7(11):e50071.
[21]Starowicz K,Makuch W,Osikowicz M,et al.Spinal anandamide produces analgesia in neuropathic rats:Possible CB1-and TRPV1-mediated mechanisms[J].Neuropharmacology,2012,62(4):1746-1755.
[22]Nabissi M,Morelli M B,Santoni M,et al.Triggering of the TRPV2 channel by cannabidiol sensitizes glioblastoma cells to cytotoxic chemotherapeutic agents[J].Carcinogenesis,2013,34(1):48-57.
[23]Alessandri-Haber N,Yeh J J,Boyd A E,et al.Hypotonicity induces TRPV4-mediated nociception in rat[J].Neuron,2003,39(3):497-511.
[24]杨全,程轩轩,郭楚楚,等.广金钱草种子的化学成分和DPPH自由基清除活性研究[J].中草药,2015,46(17):2517-2521.
[2]Campbell S T,Franks C E,Borne A L,et al.Chemoproteomic discovery of a ritanserin-targeted kinase network mediating apoptotic cell death of lung tumor cells[J].Mol Pharmacol,2018,94(5):1246-1255.
[3]程彬峰,侯媛媛,姜民,等.基于网络药理学的清肺消炎丸抗炎机制的初步研究[J].药学学报,2013,48(5):686-693.
[4]刘学,崔健,陈新.广金钱草现代研究进展[J].长春中医药大学学报,2006,22(4):84-85.
[5]刘茁,董焱,王宁,等.金钱草的化学成分[J].沈阳药科大学学报,2005,22(6):422-437.
[6]陈丰连,王术玲,徐鸿华.广金钱草挥发油的气相色谱-质谱分析[J].广州中医药大学学报,2015,22(4):302-303.
[7]刘瑞来,刘俊劭,林志銮,等.不同方法提取的金钱草叶挥发油化学成分的GC-MS分析[J].安徽农业科学,2012,40(12):7036-7037.
[8]Spitzer R,Jain A N.Surflex-Dock:Docking benchmarks and real-world application[J].J Comput Aided Mol Des,2012,26(6):687-699.
[9]Xie C,Mao X,Huang J,et al.KOBAS 2.0:A web server for annotation and identification of enriched pathways and diseases[J].Nucleic Acids Res,2011,doi:10.1093/nar/gkr483.
[10]Dainaa A,Michielin O,Zoete V.SwissADME:A free web tool to evaluate pharmacokinetics,drug-likeness and medicinal chemistry friendliness of small molecules[J].Sci Rep,2017,doi:10.1038/srep42717.
[11]林桂源,姚华聪,郑细娜,等.基于分子对接技术的常用降糖中药有效成分虚拟筛选[J].中国实验方剂学杂志,2015,21(15):202-206.
[12]Berman H M,Westbrook J,Feng Z,et al.The protein data bank[J].Nucleic Acids Res,2000,28(1):235-242.
[13]Lishko P V,Procko E,Jin X,et al.The ankyrin repeats of trpv1 bind multiple ligands and modulate channel sensitivity[J].Neuron,2007,54(6):905-918.
[14]Inada H,Procko E,Sotomayor M,et al.Structural and biochemical consequences of disease-causing mutations in the ankyrin repeat domain of the human TRPV4channel[J].Biochemistry,2012,51(31):6195-6206.
[15]Mc Cleverty C J,Koesema E,Patapoutian A,et al.Crystal structure of the human TRPV2 channel ankyrin repeat domain[J].Protein Sci,2006,15(9):2201-2206.
[16]Sutton R B,Sprang S R.Structure of the protein kinase Cbeta phospholipid-binding C2 domain complexed with Ca2+[J].Structure,1998,6(11):1395-1405.
[17]Shannon P,Markiel A,Ozier O,et al.Cytoscape:Asoftware environment for integrated models of biomolecular interaction networks[J].Genome Res,2003,13(11):2498-2504.
[18]Holzer P.Transient receptor potential(TRP)channels as drug targets for diseases of the digestive system[J].Pharmacol Ther,2011,131(1):142-170.
[19]Steinberg S F.Cardiac actions of protein kinase Cisoforms[J].Physiology,2012,27(3):130-139.
[20]Frazier W J,Xue J,Luce W A,et al.MAPK signaling drives inflammation in LPS-stimulated cardiomyocytes:The route of crosstalk to G-protein-coupled receptors[J].PLoS One,2012,7(11):e50071.
[21]Starowicz K,Makuch W,Osikowicz M,et al.Spinal anandamide produces analgesia in neuropathic rats:Possible CB1-and TRPV1-mediated mechanisms[J].Neuropharmacology,2012,62(4):1746-1755.
[22]Nabissi M,Morelli M B,Santoni M,et al.Triggering of the TRPV2 channel by cannabidiol sensitizes glioblastoma cells to cytotoxic chemotherapeutic agents[J].Carcinogenesis,2013,34(1):48-57.
[23]Alessandri-Haber N,Yeh J J,Boyd A E,et al.Hypotonicity induces TRPV4-mediated nociception in rat[J].Neuron,2003,39(3):497-511.
[24]杨全,程轩轩,郭楚楚,等.广金钱草种子的化学成分和DPPH自由基清除活性研究[J].中草药,2015,46(17):2517-2521.