青翘不同极性部位抗炎作用的谱效关系分析

Spectrum-effect Relationship of Anti-inflammatory Effect of Different Polar Parts of Unripe Forsythiae Fructus

目的:建立青翘水提物及其不同极性部位的指纹图谱,阐明各部位共有峰所代表的化学成分对抗炎药效贡献的大小,揭示其药效物质基础。方法:采用HPLC建立青翘各部位的指纹图谱,流动相乙腈-0.1%甲酸水溶液梯度洗脱,检测波长235 nm。通过青翘不同部位样品对急性肺损伤模型大鼠3个药效评价指标肿瘤坏死因子-α(TNF-α),白细胞介素-6(IL-6)及IL^(-1)β的抑制作用来反映其抗炎活性,利用灰色关联与双变量相关分析方法建立谱效关系。结果:灰色关联分析得出7个对药效贡献较大的峰,分别为23,25,31,29,24,26,30号峰。双变量相关分析显示23号峰含量变化与TNF-α呈极显著负相关;18,13号峰含量变化与IL-6呈极显著和显著性负相关;22号峰含量变化与IL^(-1)β呈极显著性负相关;7,13和6号等峰含量变化与IL^(-1)β呈显著负相关。这些色谱峰所代表的化学成分含量越高,降低TNF-α,IL-6及IL^(-1)β的效果越显著,发挥的抗炎效果越好。结论:23,13,25和31号等色谱峰对青翘抗炎药效的贡献最大。建立的HPLC指纹图谱方法简便、重复性良好。初步证明青翘的抗炎药效物质主要集中在乙酸乙酯部位,为中等极性的成分,抗炎作用是这些成分共同作用的结果。

Objective： To establish high performance liquid chromatography （HPLC） fingerprints of aqueous extract of unripe Forsythiae Fructus and its different polar parts, to clarify the contribution of fingerprints peaks in different parts of unripe Forsythiae Fructus to inhibit inflammation, and to reveal the pharmacodynamic material basis. Method： HPLC was introduced to analyze fingerprint of different parts of unripe Forsythiae Fructus with the mobile phase consisted of acetonitrile and water including 0. 1% formic acid for gradient elution. The anti- inflammatory activity of different polar parts of unripe Forsythiae Fructus was evaluated via the variety of tumor necrosis factor-c~ （TNF-a）, interleukin-6 （IL-6） and IL-I~ concentrations of the lipopolysaccharide-induced acute lung injury model in rats. Grey relational analysis and bivariate correlation analysis were used to establish the spectrum-effect relationship. Result： Seven components with greater contribution rate of effect were obtained by grey relational analysis, the different peaks ranked as No. 23, 25, 31, 29, 24, 26 and 30. Bivariate correlation analysis showed that there was a significant negative correlation between the No. 23 peak content and TNF-α, between the No, 18 and 13 peaks contents with IL-6, between No. 22 peak content with IL-1β, between the No. 7, 13 and 6 peaks contents with IL-1β. That was to say, the higher contents of chemical components of these peaks, the lower of TNF-α, IL-6 and IL-1β, and the better was the anti-inflammatory effect. Conclusion： According to the results of two kinds of analysis, it is concluded that No. 23, 13, 25, 31 and other peaks have the greatest contribution to the anti-inflammatory effect of unripe Forsythiae Fructus. The established HPLC fingerprint method is simple and reproducible. The anti-inflammatory substances of unripe Forsythiae Fructus mainly concentrate in ethyl acetate part, which is the middle polar composition. Moreover, the anti-inflammatory effect is the combined action of these components.