地菍止血作用的谱效关系研究及体外凝血活性初探

Study on the Spectral Effect Relationship of Hemostatic Action of Melastoma dodecandrum Herb and In Vitro Coagulation Activity

目的:建立地菍50%乙醇提取物中正丁醇部位的HPLC指纹图谱,探讨其与止血作用的谱效关系及体外凝血活性,阐明其药效物质基础。方法:采用HPLC法建立10批地菍50%乙醇提取物中正丁醇部位的指纹图谱;以毛细玻管法测定小鼠凝血时间,大鼠断尾法测定体外凝血效果;用灰色关联度分析法和偏最小二乘回归分析法关联药理作用数据与指纹图谱共有峰,分析地菍的谱效关系。结果:地菍50%乙醇提取物中正丁醇部位能明显缩短凝血时间,并具有体外凝血作用;建立的指纹图谱有14个共有峰,指认了没食子酸、牡荆素、芦丁3个成分;各共有峰与止血作用灰色关联分析结果的大小顺序为色谱峰5>9>4>1>7>6>14>10>13>12>2>11>3>8;偏最小二乘回归分析中共有峰1、3~13与止血时间呈负相关,且色谱峰1、3、4、7~9、11、12的VIP值>1。结论:该研究为地菍质量标准提升以及进一步药效物质研究提供参考。

Objective:To establish HPLC fingerprint of n-butanol extract from the 50%ethanol extract of Melastoma dodecandrum herb,to explore the spectrum-effect relationship between it and hemostasis and in vitro coagulation activity,clarify the pharmacodynamic substance basis.Methods:HPLC was used to establish the fingerprint of 10 batches of n-butanol extract from the 50%ethanol extract of Melastoma dodecandrum herb.The coagulation time of mice was determined by capillary tube method and the coagulation effect in vitro was determined by cutting tail in rats.Grey relational analysis and partial least square regression analysis were used to correlate the pharmacological action data and common peaks of fingerprint,and to analyze the spectral effect relationship of Melastoma dodecandrum herb.Results:The n-butanol extract from the 50%ethanol extract of Melastoma dodecandrum herb could significantly shorten the coagulation time and had the effect of coagulation in vitro.There were 14 common peaks in HPLC fingerprints and gallic acid,vitexin and rutin were identified.The order of grey correlation analysis between common peaks and hemostasis was chromatographic peak 5>9>4>1>7>6>14>10>13>12>2>11>3>8.Partial least square regression analysis showed that peaks 1,3 to 13 were negatively correlated with hemostatic time,and the VIP values of chromatographic peaks 1,3,4,7 to 9,11,12 were greater than 1.Conclusion:This study provides reference for the improvement of local quality standards and further research on pharmacodynamic substances of Melastoma dodecandrum.