通脉方中异黄酮类成分清除DPPH自由基的活性研究

摘要

Objective:Tongmai formula(TMF)is a drug combination of three components including Puerariae Lobatae Radix [roots of Pueraria lobata(Willd.)Ohwi],Salviae Miltiorrhizae Radix(roots of Salvia miltiorrhiza Bge.)and Chuanxiong Rhizoma(rhizomes of Ligusticum chuanxiong Hort.) in a weight ratio of 1 ∶ 1 ∶ 1.The scavenging activity against DPPH free-radical by isoflavonoid compounds isolated from TMF were studied in this paper.The assayed isoflavonoid compounds include daidzein,formononetin,5-hydroxylononin,ononin,daidzin,3'-methoxypuerarin,genistin,puerarin,formononetin8-C-β-D-apiofuranosyl-(1→6)-O-β-D-glucopyranoside,formononetin-7-O-β-D-apiofuranosyl-(1→6)-O-β-D-glucopyranoside,lanceolarin,kakkanin,daidzein-7,4'-di-O-β-D-glucopyranoside,mirificin,3'-hydroxypuerarin,3′-methoxydaidzin,form-ononetin-8-C-β-D-xylopyranosyl-(1→ 6)-O-β-D-glucopyranoside,genistein-8-C-β-D-apiofuranosyl-(1→ 6)-O-β-D-glucopyranoside,genistein-7-O-β-D-apiofuranosyl-(1 → 6)-O-β-D-glucopyranoside (ambocin),3'-hydroxymirificin,6"-O-β-D-xylosylpuerarin,biochanin-A-8-C-β-D-apiofuranosyl-(1 → 6)-O-β-D-glucopyranoside,3'-methoxydaidzein-7,4'-di-O-β-D-glucopyranoside,daidzein-7-O-β-D-glucopyranosyl-(1 →4)-O-β-D-glucopyranoside,and daidzein-7-O-α-D-glucopyranosyl-(1 →4)-O-β-D-glucopyranoside.Methods:The in vitro antioxidant potentials of above-mentioned 25 isoflavonoid compounds were evaluated by DPPH(2,2'-diphenyl-1-picrylhydrazyl)free-radical scavenging test.Results:In vitro antioxidant studies showed that 3'-hydroxypuerarin,3'-hydroxymirificin,3'-methoxypuerarin,and 3'-methoxydaidzin in TMF possessed notable antioxidant effects on DPPH free-radical scavenging in a concentration-dependent manner and their values of IC50 were (9.9 ± 0.6),(13.7 ± 1.4),(57.3 ± 1.2),and(66.4 ±2.3) μmol ·L-1,respectively.While the IC50 values of butylated hydroxytoluene and ascorbic acid used as the positive control under the same condition were (75.4 ± 6.9) and (16.5 ± 1.6) μmol · L-1,respectively.Conclusion:The study result could predict that 3′-hydroxypuerarin,3′-hydroxymirificin,3'-methoxypuerarin,and 3'-methoxydaidzin may be an important part of the bioactive material basis of TMF.%目的:通脉方是由葛根、丹参和川芎3味药按质量1∶1∶1组成的复方.本文研究通脉方中3′-羟基葛根素、3′-甲氧基葛根素、3′-甲氧基大豆苷、3′-羟基泰国野葛根素等25种异黄酮类化合物清除DPPH(2,2′-diphenyl-1-picrylhydrazyl)自由基的活性.方法:通过DPPH自由基清除实验,对25个异黄酮类化合物的体外抗氧化活性进行测定.结果:体外抗氧化研究表明,通脉方中的3′-羟基葛根素、3′-羟基泰国野葛根素、3′-甲氧基葛根素和3'-甲氧基大豆苷具有清除DPPH自由基的活性,呈浓度依赖性,它们的IC50值分别为(9.9±0.6)、(13.7±1.4)、(57.3±1.2)和(66.4±2.3) μmol·L-1.在同样条件下,阳性对照药丁羟甲苯和抗坏血酸的IC50值分别为(75.4±6.9)和(16.5±1.6) μmol ·L-1.结论:本研究结果能够预测3′-羟基葛根素、3'-羟基泰国野葛根素、3′-甲氧基葛根素和3′-甲氧基大豆苷可能是通脉方生物活性物质基础的重要组成部分.