当归地上部分补血作用的谱效关系研究
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摘要
采用液相色谱质谱(LC-MS)方法建立10个不同产地的当归地上部分(AAS)色谱指纹图谱;用乙酰苯肼(APH)复制血虚小鼠模型,观察AAS水提液对血虚小鼠G_6PD,GR,IL-2的影响;通过偏最小二乘(partial least squares,PLS)回归分析法将LC-MS色谱指纹图谱中各共有峰的相对含量和补血药效数据相关联,研究谱效相关性。结果显示,AAS高、中、低3个剂量组均有一定的补血作用(P<0.05),高剂量组补血效果最好(P<0.01)。根据PLS回归系数大小确定了AAS中各共有峰对补血作用的贡献程度。其中,7个共有色谱峰对补血作用的贡献较大,分别为P17(未知),P18(未知),P19(未知),P28(23-乙酰泽泻醇B或其异构体),N5(木犀草素),N11(1-咖啡酰奎宁酸)及N14(未知)。该实验研究了AAS的LC-MS色谱指纹图谱与补血作用之间的相关性,确定了AAS补血的有效成分,为当归资源的综合开发利用奠定了一定的理论基础。
Ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to establish the chromatography fingerprint for aerial parts of Angelica sinenis(AAS) from 10 different places. Acetyl-phenyl-hydrazine(APH) was used to duplicate the mouse model of blood deficiency. Then partial least squares regression was used to investigate the spectrum-effect relationship between the relative contents and the data of enriching blood pharmacodynamics efficacy. The results showed that the three groups of high, medium and low doses of AAS had certain enriching blood activities(P<0.05), and the high dose group had the best effect(P<0.01). The contribution degree of the AAS to enriching blood activities of each common peaks were determined by PLS regression coefficient. Among them, 7 common peaks, including P17(unknown), P18(unknown), P19(unknown), P28(alisol B 23-acetate or its isomer), N5(luteolin), N11(1-caffeoylquinicacid,1-O-caffeoylquinic acid) and N14(unknown), contributed significantly to the effect of enriching blood activities. This paper dealed with the investigation on the spectrum-effect relationship between enriching blood activities and LC-MS chromatography fingerprint of AAS, and determination of the effective compositions of AAS with enriching blood activities. It provided theoretical foundation for the comprehensive development and utilization of AAS.
Ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to establish the chromatography fingerprint for aerial parts of Angelica sinenis(AAS) from 10 different places. Acetyl-phenyl-hydrazine(APH) was used to duplicate the mouse model of blood deficiency. Then partial least squares regression was used to investigate the spectrum-effect relationship between the relative contents and the data of enriching blood pharmacodynamics efficacy. The results showed that the three groups of high, medium and low doses of AAS had certain enriching blood activities(P<0.05), and the high dose group had the best effect(P<0.01). The contribution degree of the AAS to enriching blood activities of each common peaks were determined by PLS regression coefficient. Among them, 7 common peaks, including P17(unknown), P18(unknown), P19(unknown), P28(alisol B 23-acetate or its isomer), N5(luteolin), N11(1-caffeoylquinicacid,1-O-caffeoylquinic acid) and N14(unknown), contributed significantly to the effect of enriching blood activities. This paper dealed with the investigation on the spectrum-effect relationship between enriching blood activities and LC-MS chromatography fingerprint of AAS, and determination of the effective compositions of AAS with enriching blood activities. It provided theoretical foundation for the comprehensive development and utilization of AAS.
引文
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[24] 杨晓,杨钊,李新荣. LC-MS/MS法同时测定维C银翘片中6种成分[J].中成药,2013,35(10):2154.
[25] 张丽君,杨敏,刘君怡,等.LC-MS/MS同时测定桑杏汤水提液中5种成分的含量[J].时珍国医国药,2016,27(3):561.
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[27] 朱云祥,陈璐琳,龚婧如,等.LC-Q-TOF-MS及LC-IT-MS分析酸枣仁汤的化学成分[J].中国中药杂志,2014,39(3):459.
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[32] 刘立,段金廒,唐于平.当归-红花配伍化学成分UHPLC-Q-TOF-MS分析及配伍协同增效作用研究[J].中华中医药杂志,2017,32(2):999.
[33] 张倩.基于LC-MSn技术的清开灵注射液绿原酸类成分研究[D].北京:北京中医药大学,2013.
[34] 元超,王鸿发,胡璇,等.UPLC-Q-TOF-MSE技术快速定性艾纳香抗菌有效部位化学成分[J].天然产物研究与开发,2018,30(11):1904.
[35] 李伟霞,唐于平,尚尔鑫,等.当归、川芎和归芎药对中主要芳香酸类成分在血虚大鼠体内的代谢研究[J].中华中医药杂志,2013,28(5):1212.
[36] 彭贤,黄舒,郦皆秀,等.泽泻属植物化学成分与药理活性[J].国外医药·植物药分册,2000(6):245.
[37] 靳如娜,齐晓甜,孙蓉,等.经典名方当归补血汤的药效物质基础和质量控制研究进展[J].中国实验方剂学杂志,doi:10.13422/j.cnki.syfjx.20190648.
[38] 杨英来,崔方,胡芳,等.当归补血、活血作用的谱效关系研究[J].中国中药杂志,2013,38(22):3923.
[2] 秦甜,刘建新,周小青,等.从血虚证探讨证的认识论特征[J].环球中医药,2014, 7(8):612.
[3] 张文卓,董慧,黄晓巍.血虚证中医药研究进展[J].中国当代医药,2013,20(1):16.
[4] 钱宏梁,潘志强.中医血虚证及其动物模型制备方法评析[J].广州中医药大学学报,2018, 35(1):176.
[5] 谢东杰,王爱迪,刘宝山.当归补血汤在血液疾病中作用机制的研究进展[J].中华中医药杂志,2018,33(6):2489.
[6] 黄丽萍,陈耀辉,吴素芬,等.当归补血汤含药血清对血虚小鼠骨髓造血功能的影响[J].中药药理与临床,2014,30(2):18.
[7] 陈耀辉,黄丽萍,吴素芬,等.当归补血汤与人参当归方、灵芝阿胶方的补气生血作用比较[J].中药药理与临床,2015,31(1):25.
[8] 贾梅,郑传柱,张丽,等.胶艾汤与四物汤对血虚模型大鼠补血功效的比较研究[J].中草药,2015,46(11):1641.
[9] 杨明会,马增春,窦永起,等.四物汤对血虚证患者血清蛋白质的影响[J].中国中药杂志,2008,33(4):420.
[10] 何瑶,傅超美,毛茜,等.四物汤不同提取工艺对血虚模型小鼠造血功能的影响[J].中国实验方剂学杂志,2012,18(12):199.
[11] 黄琪,雷鹏,李新中,等.四物汤配方颗粒汤剂治疗血虚证临床疗效观察[J].时珍国医国药,2015,26(1):127.
[12] 周鸿,黄含含,张静泽,等.川芎-当归药对研究进展[J].中成药,2015,37(1):184.
[13] 黄美艳,尚尔鑫,唐于平,等.基于药物相互作用对当归-川芎配伍养血补血效应研究[J].中国中西医结合杂志,2013,33(4):520.
[14] 李伟霞,唐于平,郭建明,等.当归-川芎药对及其组成药味对3种血虚模型补血作用的比较研究[J].中国中药杂志,2011,36(13):1813.
[15] 李淑娇,唐于平,李伟霞,等.当归-红花不同配比对血虚小鼠补血作用的比较[J].中国药理学通报,2014,30(6):877.
[16] 马霞.当归不同部位的化学成分与补血作用研究[D].兰州:甘肃中医药大学, 2018:84.
[17] 曾令军,林兵,宋洪涛.中药谱效关系研究进展及关键问题控讨[J].中国中药杂志,2015,40(8):1425.
[18] 吕邵娃,董书羽,郭玉岩,等.数据分析技术在中药谱效关系中的应用进展[J].中国实验方剂学杂志,2015,21(5):228.
[19] Shi M J, Zhang Y, Song M M, et al. Screening the marker components in Psoralea corylifolia L. with the aids of spectrum-effect relationship and component knock-out by UPLC-MS2[J]. Molecules, 2018, 19(11): 3439.
[20] 王国华,张虎,魏岳嵩.偏最小二乘回归在SPSS软件中的实现[J].统计与决策,2017(7):67.
[21] Wang J M, Peng L N, Shi M J, et al. Spectrum effect relationship and component knock-out in Angelica dahurica radix by high performance liquid chromatography-Q exactive hybrid quadrupole-orbitrap mass spectrometer[J]. Molecules, 2017, 22(27): 1231.
[22] 金若敏,宁炼,陈长勋,等.血虚模型动物制备及当归补血汤的作用研究[J].中成药,2001,23(4):270.
[23] Zhang X F, Chen J,Yang J L, et al. UPLC-MS/MS analysis for antioxidant components of Lycii Fructus based on spectrum-effect relationship[J]. Talanta, 2018, 180: 392.
[24] 杨晓,杨钊,李新荣. LC-MS/MS法同时测定维C银翘片中6种成分[J].中成药,2013,35(10):2154.
[25] 张丽君,杨敏,刘君怡,等.LC-MS/MS同时测定桑杏汤水提液中5种成分的含量[J].时珍国医国药,2016,27(3):561.
[26] 王翌臣,王焕军,张玲,等.大狼把草的化学成分液质联用快速鉴定分析[J].中国实验方剂学杂志,2018,24(17):82.
[27] 朱云祥,陈璐琳,龚婧如,等.LC-Q-TOF-MS及LC-IT-MS分析酸枣仁汤的化学成分[J].中国中药杂志,2014,39(3):459.
[28] 杨帆,肖远胜,章飞芳,等.当归化学成分的HPLC-MS/MS分析[J].药学学报,2006,41(41):1080.
[29] 牛研,王书芳.LC-Q-TOF-MS和LC-IT-MSn分析当归芍药散中化学成分[J].中草药,2014,45(8):1059.
[30] Chen L L, Qi J, Chang Y X, et al. Identification and determination of the major constituents in traditional Chinese medicinal formula Danggui-Shaoyao-San by HPLC-DAD-ESI-MS/MS[J]. J Pharm Biomed Anal, 2009, 50: 130.
[31] 孙倩.基于液质联用技术的大小蓟多组分分析与黄酮类成分的药物代谢动力学研究[D].石家庄:河北医科大学,2013.
[32] 刘立,段金廒,唐于平.当归-红花配伍化学成分UHPLC-Q-TOF-MS分析及配伍协同增效作用研究[J].中华中医药杂志,2017,32(2):999.
[33] 张倩.基于LC-MSn技术的清开灵注射液绿原酸类成分研究[D].北京:北京中医药大学,2013.
[34] 元超,王鸿发,胡璇,等.UPLC-Q-TOF-MSE技术快速定性艾纳香抗菌有效部位化学成分[J].天然产物研究与开发,2018,30(11):1904.
[35] 李伟霞,唐于平,尚尔鑫,等.当归、川芎和归芎药对中主要芳香酸类成分在血虚大鼠体内的代谢研究[J].中华中医药杂志,2013,28(5):1212.
[36] 彭贤,黄舒,郦皆秀,等.泽泻属植物化学成分与药理活性[J].国外医药·植物药分册,2000(6):245.
[37] 靳如娜,齐晓甜,孙蓉,等.经典名方当归补血汤的药效物质基础和质量控制研究进展[J].中国实验方剂学杂志,doi:10.13422/j.cnki.syfjx.20190648.
[38] 杨英来,崔方,胡芳,等.当归补血、活血作用的谱效关系研究[J].中国中药杂志,2013,38(22):3923.