基于主成分分析和灰色关联分析方法评价雷公藤配伍金钱草相杀减毒作用的化学基础

摘要

Objective To investigate the chemical bases and mechanism of mutual detoxification of the compatibility of Tripterygium wilfordii (TW) with Lysimachia christina (LC).Methods This study was conducted in October 2015.Dry extracts of ethyl acetate extract (EAE),4/1 EAE,2/1 EAE,1/1 EAE,1/2 EAE,and 1/4 EAE were prepared,and then the corresponding test solutions were made up.After the linearity of triptolide (TP) and celastrol (CEL),the precision of the instrument,the stability of the test solutions,and the good reproducibility of the methods were determined,the contents of TP and CEL were calculated based on their standard curves,respectively,and the HPLC chemical characteristics (characteristic peaks) before and after compatibility of TW and LC were identified.The TP and CEL contents of each group before and after compatibility,and the peak area integral values of the unknown differential characteristic peaks were calculated by the PCA method to obtain the Z value of the chemical score.The GCA method was used to analyze the grey correlation between the peaks of TP,CEL and other chemical differences and the Z value of chemical scores,and ranked according to the gray correlation coefficient.The degree of influence of each chemical characteristic peak on the chemical score was compared.Results A total of 12 distinctive peaks were identified under the TP and CEL conditions,naming as TP,CEL,and 1-10 peaks.After being dimensionally reduced by PCA,two principal components were produced.Then the weighted content of TP-CEL was obtained by weighting the TP and CEL content based on the extracted information,that is,the common factor variance.The TP-CEL of 4/1 EAE,2/1 EAE,1/1 EAE,1/2 EAE,and 1/4 EAE was lower than that of EAE (P＜0.05);the TP-CEL of 2/1 EAE was lower than that of 4/1 EAE,1/1 EAE and 1/2 EAE (P＜0.05);the TP-CEL of 1/4 EAE was lower compared with that of 4/1 EAE,2/1 EAE,1/1 EAE,and 1/2 EAE (P＜0.05).The Z value of the chemical scores of 4/1 EAE,2/1 EAE,1/1 EAE, 1/2 EAE,and 1/4 EAE was lower than that of EAE (P＜0.05);the chemical score Z of 2/1 EAE was lower compared with that of 4/1 EAE,1/ 1 EAE and 1/2 EAE (P＜0.05);1/4 EAE group had lower chemical score Z than 4/1 EAE,2/1 EAE, 1/1 EAE and 1/2 EAE (P＜0.05).Based on the GCA analysis performed on the differential characteristic peaks and the Z value of the chemical scores,the gray correlation coefficients of the characteristic peaks were obtained and ranked according to the grey correlation coefficient.Ranked No.1 was No.3 characteristic peak,and TP and CEL were ranked as No.8 and No.9 of the 12 characteristic peaks,respectively.Conclusion The 12 differential chemical components found were the main chemical basis for the mutual detoxification of the compatibility of TW with LC.The mass ratio of 4/1 to 1/4 can be used to attenuate the toxicity by reducing the chemical score,and especially 1/4 is the best ratio,and its attenuation may be related to the combined effects of the chemical constituents of TP,CEL,No.1-10 characteristic peaks,in particular,the No.3 characteristic component has the greatest influence.%目的 阐释雷公藤(LGT)与金钱草(JQC)配伍相杀减毒的化学基础,初步评价其相杀减毒作用.方法 2015年10月,首先制备乙酸乙酯提取物(EAE)、4/1 EAE、2/1 EAE、1/1 EAE、1/2 EAE、1/4 EAE的干浸膏以配置相应的供试品溶液,在确定雷公藤甲素(TP)和雷公藤红素(CEL)线性、仪器精密度、供试品溶液稳定性、方法重复性良好的基础上,根据TP和CEL标准曲线分别计算TP和CEL含量,寻找LGT与JQC配伍前后高效液相色谱法(HPLC)化学差异特征成分(特征峰),将配伍前后各组的TP和CEL含量以及未知差异特征峰峰面积积分值,经过主成分分析(PCA)计算化学得分Z值.将TP、CEL及其他化学差异特征峰均与化学得分Z值进行灰色关联分析(GCA)方法,并按照灰色关联度大小进行排序,比较各化学差异特征峰对化学得分的影响程度.结果 TP和CEL测定条件下差异特征峰共有12个,分别为TP、CEL、1～10号峰.将12个差异特征峰经PCA降维,产生2个主成分.根据提取的信息量即公因子方差对TP和CEL含量进行加权,可得二者的加权含量(TP-CEL).4/1 EAE、2/1 EAE、1/1 EAE、1/2 EAE、1/4 EAE的TP-CEL均低于EAE(P＜0.05);2/1 EAE的TP-CEL低于4/1 EAE组、1/1 EAE组、1/2 EAE(P＜0.05);1/4 EAE的TP-CEL低于4/1 EAE、2/1 EAE、1/1 EAE、1/2 EAE(P＜0.05).4/1 EAE、2/1 EAE、1/1 EAE、1/2 EAE、1/4 EAE化学得分Z值低于EAE(P＜0.05);2/1 EAE化学得分Z值低于4/1 EAE、1/1 EAE、1/2 EAE(P＜0.05);1/4 EAE组化学得分Z值低于4/1 EAE、2/1 EAE、1/1 EAE、1/2 EAE(P＜0.05).将差异特征峰与化学得分Z值进行GCA分析,得各特征峰的灰色关联系数,并按灰色关联系数进行排序.排在第1的是3号特征峰,TP和CEL分别排在12个特征峰中的第8和第9.结论 寻找到的12个差异化学成分为LGT配伍JQC相杀减毒的主要化学基础,二者在质量配比4/1～1/4均可通过降低化学得分从而发挥相杀减毒作用,其中尤以1/4为最佳配比,其减毒作用可能与TP、CEL、1～10号特征化学成分的综合作用有关,其中尤以3号特征成分的影响最大.