蓬莪术醋制前后UFLC-Q-TOF-MS色谱峰与抑制小鼠尾血栓形成作用的相关性研究
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摘要
蓬莪术的生品及醋制品均为中医临床的常用药物,抑制小鼠尾血栓形成是反映中药活血化瘀作用的重要指标之一。传统炮制理论认为醋性味酸苦微温,作为辅料渗入莪术进行炮制后,可改变药物的活血化瘀作用。为此,该实验使用了中药谱效学的研究方法,通过超快速液相色谱-四极杆飞行时间质谱(UFLC-Q-TOF-MS)法建立蓬莪术生品与醋制品抑制小鼠尾血栓形成作用的谱效关系,以探讨莪术醋制后活血化瘀作用发生改变的原因。实验以蓬莪术生品与醋制品醇提物的UFLC-Q-TOF-MS图谱的峰面积及其抑制小鼠尾血栓形成药效为基础,采用OPLS-DA法对UFLC-Q-TOF-MS图谱和抑制小鼠尾血栓形成的药效进行相关性分析。经Simca-P软件分析表明指纹图谱中的16,24(去氢木香内酯),3,11,22(α-脱氢姜黄烯),19[(R)-(-)-α-姜黄烯],23,10号峰对区分蓬莪术生品及醋制品贡献较大。该研究从抑制小鼠尾血栓形成的药效角度,用谱效关系的技术手段找到了导致蓬莪术生品与醋制品药效变化的指标性成分,为阐明中药在炮制前后产生活血化瘀作用变化提供新的依据。
Both raw and vinegar products of the rhizome of Curcuma phaeocaulis are common drugs for promoting blood circulation and removing blood stasis in traditional Chinese medicine,which could be reflected in the inhibition of tail thrombosis in mice. As the traditional processing theory instructs,vinegar tastes sour and bitter,but can activate blood circulation and remove stasis after being infiltrated into the rhizome of C. phaeocaulis as an excipient. In this study,under the help of the ultrafast liquid chromatography-quadrupole time-offlight mass spectrometry( UFLC-Q-TOF-MS),the spectrum-effect relationship between the inhibition of tail thrombosis in mice and the rhizome of C. phaeocaulis both before and after the vinegar processing,were established to explore the functional changes of blood circulation and stasis after vinegar process. Based on the peak area from the fingerprint of UFLC-Q-TOF-MS of the alcohol extracts from the raw and vinegar-processed rhizome of C. phaeocaulis and their efficacy for inhibiting tail thrombosis,the correlation between the chromatography of UFLC-Q-TOF-MS and the inhibition of tail thrombosis in mice were analyzed by orthogonal partial least squares discriminant analysis( OPLS-DA) method. The results,produced by Simca-P software,showed that effective components consisted of eight peaks 16,24( aromadendrene oxide),3,11,22( dehydro-α-curcumene),19[( R)-(-)-α-curcumene],23 and 10 from the fingerprint,making great contribution to distinguish C. phaeocaulis raw products and the corresponding vinegar processed products. Therefore,from the perspective of inhibiting the formation of tail thrombosis in mice,the marker components could be found through the spectrum-effect relationship to distinguish C.phaeocaulis raw and vinegar products. This study provided new basis to explain the difference between the raw and the processed products of traditional Chinese medicine in the functional change of promoting blood circulation and removing blood stasis.
Both raw and vinegar products of the rhizome of Curcuma phaeocaulis are common drugs for promoting blood circulation and removing blood stasis in traditional Chinese medicine,which could be reflected in the inhibition of tail thrombosis in mice. As the traditional processing theory instructs,vinegar tastes sour and bitter,but can activate blood circulation and remove stasis after being infiltrated into the rhizome of C. phaeocaulis as an excipient. In this study,under the help of the ultrafast liquid chromatography-quadrupole time-offlight mass spectrometry( UFLC-Q-TOF-MS),the spectrum-effect relationship between the inhibition of tail thrombosis in mice and the rhizome of C. phaeocaulis both before and after the vinegar processing,were established to explore the functional changes of blood circulation and stasis after vinegar process. Based on the peak area from the fingerprint of UFLC-Q-TOF-MS of the alcohol extracts from the raw and vinegar-processed rhizome of C. phaeocaulis and their efficacy for inhibiting tail thrombosis,the correlation between the chromatography of UFLC-Q-TOF-MS and the inhibition of tail thrombosis in mice were analyzed by orthogonal partial least squares discriminant analysis( OPLS-DA) method. The results,produced by Simca-P software,showed that effective components consisted of eight peaks 16,24( aromadendrene oxide),3,11,22( dehydro-α-curcumene),19[( R)-(-)-α-curcumene],23 and 10 from the fingerprint,making great contribution to distinguish C. phaeocaulis raw products and the corresponding vinegar processed products. Therefore,from the perspective of inhibiting the formation of tail thrombosis in mice,the marker components could be found through the spectrum-effect relationship to distinguish C.phaeocaulis raw and vinegar products. This study provided new basis to explain the difference between the raw and the processed products of traditional Chinese medicine in the functional change of promoting blood circulation and removing blood stasis.
引文
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[15]刘彦霞,赵肖萌,温宗妍,等.角叉菜胶两种给药方式制备小鼠尾部血栓模型比较[J].中西医结合心脑血管病杂志,2013,11(7):849.
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[19] Wongravee K,Heinrich N,Holmboe M,et al. Variable selection using iterative reformulation of training set models for discrimination of samples:application to gas chromatography/mass spectrometry of mouse urinary metabolites[J]. Anal Chem,2009,81:5204.
[2]赖艳妮,严一文,徐培平.基于系统药理学探索莪术有效成分的药理作用机制[J].中国实验方剂学杂志,2017,23(14):177.
[3]李林,陆兔林,卞慧敏,等.莪术活血化瘀有效物质研究[J].上海中医药大学学报,2004,18(3):40.
[4]张妲,金城,骆骄阳,等.姜黄素类化合物体外抗凝血与抗血栓作用研究[J].中草药,2011,42(10):2070.
[5]王秀,夏泉,许杜娟,等.莪术中莪术二酮抗凝血和抗血栓作用的实验研究[J].中成药,2012,34(3):550.
[6]霍伟,段文丽,柳军,等.β-榄香烯抗凝血溶血栓活性研究[J].亚太传统医药,2013,9(8):30.
[7]唐泽耀,宗成国,林原.莪术醇的活血化瘀活性实验研究[J].中药药理与临床,2003,19(5):15.
[8]刘会珍,陆兔林,毛春芹,等.莪术不同炮制品中倍半萜及姜黄素类成分的比较[J].中成药,2014,36(4):804.
[9]覃葆,杨海玲,蒋文兰,等.广西莪术不同炮制品挥发油的含量测定及气相色谱-质谱联用分析[J].时珍国医国药,2010,21(9):2284.
[10]陆兔林,杨光明,宋珅,等.气质联用法分析炮制对莪术挥发油成分的影响[J].中成药,2003,25(10):810.
[11]曾令军,林兵,宋洪涛.中药谱效关系研究进展及关键问题探讨[J].中国中药杂志,2015,40(8):1425.
[12]刘英,徐永莉,李力.中药谱效关系研究思路与应用概况[J].时珍国医国药,2018,29(3):677.
[13]顾黎云,杜沙莉,张磊,等.基于成分及药效差异的江苏省莪术临床应用调查分析[J].中国药房,2017,28(28):4019.
[14]顾黎云,杜沙莉,张磊,等.UFLC-Q-TOF-MS法分析蓬莪术有效成分[J].中成药,2018,40(1):137.
[15]刘彦霞,赵肖萌,温宗妍,等.角叉菜胶两种给药方式制备小鼠尾部血栓模型比较[J].中西医结合心脑血管病杂志,2013,11(7):849.
[16]罗妮妮.川产道地药材蓬莪术醋制前后药学对比研究[D].成都:成都中医药大学,2015.
[17]周雯,张丽艳,谢立敏,等.超高效液相色谱-飞行时间质谱联用分析苗药头花蓼醇提物及水提物化学成分[J].中国中药杂志,2017,42(18):3557.
[18]阿基业.代谢组学数据处理方法——主成分分析[J].中国临床药理学与治疗学,2010,15(5):481.
[19] Wongravee K,Heinrich N,Holmboe M,et al. Variable selection using iterative reformulation of training set models for discrimination of samples:application to gas chromatography/mass spectrometry of mouse urinary metabolites[J]. Anal Chem,2009,81:5204.