基于灰色关联分析法探索芍药甘草汤对癫痫小鼠脑保护作用的谱效关系

Investigation of Spectrum-effect Relationship of Brain Protection of Shaoyao Gancaotang on Epileptic Mice Based on Grey Correlation Analysis

目的:探讨芍药甘草汤物质组分对癫痫小鼠脑组织中丙二醛(MDA),超氧化物歧化酶(SOD)和三磷酸腺苷(ATP)酶水平影响的谱效关系,揭示芍药甘草汤抗癫痫作用的物质基础。方法:利用HPLC建立15批芍药甘草汤煎液的指纹图谱,流动相乙腈(A)-0. 1%磷酸水溶液(B)梯度洗脱(0~15 min,19%A;15~45 min,19%~50%A;45~46 min,50%~90%A;46~48 min,90%A),流速1. 0 mL·min^-1,检测波长237 nm。采用青霉素诱导小鼠癫痫模型,考察15批芍药甘草汤煎液对癫痫模型小鼠脑组织中MDA,SOD和ATP酶水平的影响;利用灰色关联分析法将15批芍药甘草汤煎液指纹图谱特征峰峰面积与药效学指标MDA,SOD和ATP酶活性进行关联分析,建立不同物质组分与药效学指标之间的谱效关系数学模型。结果:芍药甘草汤煎液能够提高癫痫小鼠脑组织中ATP酶和SOD的含量,降低MDA含量,且大部分具有显著性差异(P<0. 05,P<0. 01)。在15批芍药甘草汤煎液的指纹图谱中标定出了28个共有峰,其中15个特征峰。灰色关联分析法发现对MDA,SOD和ATP酶活性影响有贡献的特征峰包括2号峰(芍药内酯苷),3号峰(芍药苷),5号峰(甘草苷)等,具体排序为3号峰>6号峰>12号峰>8号峰>2号峰>5号峰>9号峰>4号峰>10号峰>7号峰>13号峰>11号峰>15号峰>1号峰>14号峰。结论:芍药甘草汤是通过多组分协同作用来影响癫痫小鼠脑组织中SOD,MDA和ATP酶活性的。将灰色关联分析法引入中药复方组分与药效之间的谱效关系相关性评价中,能客观反映中药多组分协同发挥药效作用的本质,是筛选和预测中药药效物质组分的一种有效分析方法。

Objective:To investigate the spectrum-effect relationship of effect of material components in Shaoyao Gancaotang on the levels of malondialdehyde(MDA),superoxide dismutase(SOD)and adenosine triphosphatase(ATPase)in brain tissue of epileptic mice,and to reveal the material basis of the antiepileptic effect of Shaoyao Gancaotang. Method: HPLC was used to establish the fingerprint of 15 batches of Shaoyao Gancaotang,the mobile phase was consisted of acetonitrile(A)-0. 1% phosphoric acid aqueous solution(B)for gradient elution(0-15 min,19%A;15-45 min,19%A-50%A;45-46 min,50%A-90%A;46-48 min,90%A),the flow rate was 1. 0 mL·min^-1,and the detection wavelength was 237 nm. The mouse epilepsy model was induced by penicillin,the protective effect of 15 batches of Shaoyao Gancaotang on MDA,SOD and ATPase levels in brain tissue of epileptic mice was investigated. Grey correlation analysis was used to analyze the correlation between the peak areas of characteristic peaks in the fingerprint of 15 batches of Shaoyao Gancaotang and three pharmacodynamic indexes(the activities of MDA,SOD and ATPase),and the mathematical model of spectrum-effect relationship between different material components and pharmacodynamic indexes was established. Result: Shaoyao Gancaotang could increase the contents of ATPase and SOD and reduce the content of MDA in the brain tissue of epileptic mice,and most of the differences were significant(P<0. 05,P<0. 01). There were 28 common peaks in the fingerprint of 15 batches of Shaoyao Gancaotang,and 15 of them were characteristic peaks. Grey correlation analysis found that the characteristic peaks contributed to the activities of MDA,SOD and ATPase included peak 2(albiflorin),peak 3(paeoniflorin),peak 5(liquiritin),and so on,the specific ranking was peak 3>peak 6>peak 12>peak 8>peak 2>peak 5>peak 9>peak 4>peak 10>peak 7>peak 13>peak 11>peak 15>peak 1>peak 14. Conclusion: Shaoyao Gancaotang can affect the activities of SOD,MDA and ATPase in brain tissue of epileptic mice by multi-component synergy. Simultaneously,introducing the grey correlation analysis into the correlation evaluation of the spectrum-effect relationship between components and efficacy of Chinese herbal compounds,it can objectively reflect the essence of the synergistic action of multiple components in traditional Chinese medicine(TCM),and it is an effective analysis method for screening and predicting the pharmacodynamic components of TCM.