基于谱效关系的中药虎杖抗炎活性成分探讨
|
摘要
目的:基于谱效关系研究虎杖中潜在的抗炎活性成分。方法:应用脂多糖(lipopolysaccharide,LPS)诱导小鼠腹腔巨噬细胞(RAW264. 7细胞)建立炎症模型,对虎杖95%乙醇提取物及其大孔树脂洗脱部位(30%,60%,95%乙醇洗脱物)的抗炎活性进行筛选;结合超高效液相色谱-飞行时间质谱(ultra performance liquid chromatography coupled with quadrupole timeof-flight mass spectrometry,UPLC-Q-TOF/MS)技术建立特征指纹图谱;利用偏最小二乘法(partial least squares,PLS)将各部位特征成分谱峰面积与一氧化氮(NO)抑制率进行谱效关系研究,根据变异权重系数(variable important in projection,VIP)来辨识其抗炎活性成分。结果:虎杖醇提物大孔树脂60%乙醇洗脱部位对LPS诱导的细胞炎症模型中一氧化氮生成的抑制能力最强,且表现出一定的剂量依赖关系。谱效关系研究发现,虎杖醇提物及其大孔树脂洗脱部位中3个成分包括大黄素-8-O-β-D-葡萄糖苷(E-8-G),大黄素-1-O-β-D-葡萄糖苷和大黄素-8-O-(6'-O-丙二酰基)-葡萄糖苷具有潜在的抗炎活性,并从细胞层面和分子反向寻靶研究对E-8-G的抗炎活性进行了验证。结论:本研究基于谱效关系的研究方法,从虎杖中筛选到3个潜在的抗炎活性成分,该方法有助于快速准确地发现中药活性成分(群),为研究中药药效物质基础提供新的研究思路和方法。
Objective: To study the potential anti-inflammatory active ingredient of Chinese herbal Polygoni Cuspidati Rhizoma et Radix. Method: The inflammatory model of peritoneal macrophages( RAW264. 7 cells) induced by lipopolysaccharide( LPS) in mice was used to screen out the anti-inflammatory activity of 95%ethanol extract of Polygoni Cuspidati Rhizoma et Radix and its macroporous resin elution site( 30%,60%,95%ethanol eluting site). The characteristic fingerprints were established by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry( UPLC-Q-TOF/MS) technology,and then partial least squares method( partial least squares,PLS) was used to study the spectrum-effect relationship between the peak area of the characteristic components and the inhibition rate of nitric oxide( NO),and potential anti-inflammatory active ingredients were identified according to variable important in projection( VIP). Result: The ethanol extract macroporous resin 60% ethanol elution site of Polygoni Cuspidati Rhizoma et Radix had the strongest inhibition ability of nitric oxide formation, with a certain dose-dependent relationship. The study of spectrum-effect relationship showed that 3 components had potential anti-inflammatory activity,namely Emodin-8-O-β-D-glucoside( E-8-G),Emodin-1-O-β-D-giucoside,and Emodin-8-O-( 6'-O-malonyl)-glucoside. In addition,the antiinflammatory activity of E-8-G was further validated at the cell level through molecular docking analysis.Conclusion: Three potential anti-inflammatory active ingredients were found base on the spectrum-effect relationship. This study strategy is helpful to find the active ingredients( group) of traditional Chinese medicine,and provides new research ideas and methods for studying the material basis of Chinese herbal medicine.
Objective: To study the potential anti-inflammatory active ingredient of Chinese herbal Polygoni Cuspidati Rhizoma et Radix. Method: The inflammatory model of peritoneal macrophages( RAW264. 7 cells) induced by lipopolysaccharide( LPS) in mice was used to screen out the anti-inflammatory activity of 95%ethanol extract of Polygoni Cuspidati Rhizoma et Radix and its macroporous resin elution site( 30%,60%,95%ethanol eluting site). The characteristic fingerprints were established by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry( UPLC-Q-TOF/MS) technology,and then partial least squares method( partial least squares,PLS) was used to study the spectrum-effect relationship between the peak area of the characteristic components and the inhibition rate of nitric oxide( NO),and potential anti-inflammatory active ingredients were identified according to variable important in projection( VIP). Result: The ethanol extract macroporous resin 60% ethanol elution site of Polygoni Cuspidati Rhizoma et Radix had the strongest inhibition ability of nitric oxide formation, with a certain dose-dependent relationship. The study of spectrum-effect relationship showed that 3 components had potential anti-inflammatory activity,namely Emodin-8-O-β-D-glucoside( E-8-G),Emodin-1-O-β-D-giucoside,and Emodin-8-O-( 6'-O-malonyl)-glucoside. In addition,the antiinflammatory activity of E-8-G was further validated at the cell level through molecular docking analysis.Conclusion: Three potential anti-inflammatory active ingredients were found base on the spectrum-effect relationship. This study strategy is helpful to find the active ingredients( group) of traditional Chinese medicine,and provides new research ideas and methods for studying the material basis of Chinese herbal medicine.
引文
[1]PENG W,QIN R,LI X,et al. Botany,phytochemistry,pharmacology,and potential application of Polygonum cuspidatum Sieb. et Zucc[J]. J Ethnopharmacol,2013,148(3):729-745.
[2]欧水平,任丽,王玉和,等.虎杖提取物中虎杖苷及白藜芦醇在大鼠胃肠道吸收的动力学分析[J].中国实验方剂学杂志,2018,24(3):30-35.
[3]时圣明,潘明佳,王文倩,等.虎杖的化学成分及药理作用研究进展[J].药物评价研究,2016,39(2):313-321.
[4]樊慧婷,丁世兰,林洪生.中药虎杖的药理研究进展[J].中国中药杂志,2013,38(15):2545-2548.
[5]Kim K W,Ha K T,Park C S,et al. Polygonum cuspidatum,compared with baicalin and berberine,inhibits inducible nitric oxide synthase and cyclooxygenase-2 gene expressions in RAW 264. 7macrophages[J]. Vascul Pharmacol,2007,47(2/3):99-107.
[6]Ghanim H,Sia C L, Abuaysheh S, et al. An antiinflammatory and reactive oxygen species suppressive effects of an extract of Polygonum cuspidatum containing resveratrol[J]. J Clin Endocrinol Metab,2010,95(9):E1-E8.
[7]Bralley E E,Greenspan P,Hargrove J L,et al. Topical anti-inflammatory activity of Polygonum cuspidatum extract in the TPA model of mouse ear inflammation[J].J Inflamm,2008,5(1):1-8.
[8]HAN J H,Koh W,Lee H J,et al. Analgesic and antiinflammatory effects of ethyl acetate fraction of Polygonum cuspidatum in experimental animals[J].Immunopharmacol Immunotoxicol, 2012, 34(2):191-195.
[9]张青,谭鹏,罗秋月,等. UPLC在中药及其复方分析中的应用和研究进展[J].中国实验方剂学杂志,2018,24(20):22-30.
[10]吉日木巴图,范娜,王蕊,等.基于分子对接技术探讨残黄片退黄作用机制[J].中国实验方剂学杂志,2018,24(24):1-8.
[11]董静,王弘,万乐人,等.高效液相色谱/电喷雾-离子阱-飞行时间质谱分析鉴定中药虎杖中的主要化学成分[J].色谱,2009,27(4):425-430.
[12]李菁雯,陈祥龙,孟祥智.虎杖及其提取物的研究进展[J].中医药学报,2011,39(3):103-106.
[13]徐新,李德明,徐佳薇,等.虎杖提取物的抗炎作用及对一氧化氮生成的影响[J].中国医院药学杂志,2013,33(16):1311-1315.
[14]贾玉梅,王君明,崔瑛.基于二苯乙烯类为主要活性成分的虎杖药理作用研究进展[J].中国实验方剂学杂志,2011,17(9):263-269.
[15]肖遂.基于谱效关系的中药铁苋菜抑菌物质辨识方法研究[D].北京:中国农业科学院,2013.
[2]欧水平,任丽,王玉和,等.虎杖提取物中虎杖苷及白藜芦醇在大鼠胃肠道吸收的动力学分析[J].中国实验方剂学杂志,2018,24(3):30-35.
[3]时圣明,潘明佳,王文倩,等.虎杖的化学成分及药理作用研究进展[J].药物评价研究,2016,39(2):313-321.
[4]樊慧婷,丁世兰,林洪生.中药虎杖的药理研究进展[J].中国中药杂志,2013,38(15):2545-2548.
[5]Kim K W,Ha K T,Park C S,et al. Polygonum cuspidatum,compared with baicalin and berberine,inhibits inducible nitric oxide synthase and cyclooxygenase-2 gene expressions in RAW 264. 7macrophages[J]. Vascul Pharmacol,2007,47(2/3):99-107.
[6]Ghanim H,Sia C L, Abuaysheh S, et al. An antiinflammatory and reactive oxygen species suppressive effects of an extract of Polygonum cuspidatum containing resveratrol[J]. J Clin Endocrinol Metab,2010,95(9):E1-E8.
[7]Bralley E E,Greenspan P,Hargrove J L,et al. Topical anti-inflammatory activity of Polygonum cuspidatum extract in the TPA model of mouse ear inflammation[J].J Inflamm,2008,5(1):1-8.
[8]HAN J H,Koh W,Lee H J,et al. Analgesic and antiinflammatory effects of ethyl acetate fraction of Polygonum cuspidatum in experimental animals[J].Immunopharmacol Immunotoxicol, 2012, 34(2):191-195.
[9]张青,谭鹏,罗秋月,等. UPLC在中药及其复方分析中的应用和研究进展[J].中国实验方剂学杂志,2018,24(20):22-30.
[10]吉日木巴图,范娜,王蕊,等.基于分子对接技术探讨残黄片退黄作用机制[J].中国实验方剂学杂志,2018,24(24):1-8.
[11]董静,王弘,万乐人,等.高效液相色谱/电喷雾-离子阱-飞行时间质谱分析鉴定中药虎杖中的主要化学成分[J].色谱,2009,27(4):425-430.
[12]李菁雯,陈祥龙,孟祥智.虎杖及其提取物的研究进展[J].中医药学报,2011,39(3):103-106.
[13]徐新,李德明,徐佳薇,等.虎杖提取物的抗炎作用及对一氧化氮生成的影响[J].中国医院药学杂志,2013,33(16):1311-1315.
[14]贾玉梅,王君明,崔瑛.基于二苯乙烯类为主要活性成分的虎杖药理作用研究进展[J].中国实验方剂学杂志,2011,17(9):263-269.
[15]肖遂.基于谱效关系的中药铁苋菜抑菌物质辨识方法研究[D].北京:中国农业科学院,2013.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |