基于含药血清经时谱效关系的茵陈蒿汤保肝作用药效物质研究

目的：阐明茵陈蒿汤的保肝作用药效物质。方法：以SD大鼠为供体制备茵陈蒿汤灌胃给药后10个时间点含药血清，高效液相色谱法（HPLC）测定含药血清指纹图谱，采用HL-7702肝细胞损伤模型对含药血清进行保肝作用药效评价，分别采用药时曲线和时效曲线比较及双变量相关性分析的谱-效关系分析方法指认含药血清指纹图谱中与药效相关的色谱峰。结果：药时曲线和时效曲线比较发现，含药血清指纹图谱中4~7、20号峰对损伤肝细胞具有明显保护作用，8、11~19号峰对损伤肝细胞具有保护作用，但可能也具有一定肝细胞毒性有关，1~3及9号峰具有肝细胞毒性；双变量相关性分析发现，含药血清指纹图谱中1~9、15~17、20号峰对损伤肝细胞具有一定保护作用，其中4、5、20号峰活性较强，6号峰活性最强。结论：茵陈蒿汤保肝作用的药效物质主要包括来源于大黄的蒽醌类成分、来源于栀子的环烯醚萜类成分和西红花酸类成分及以上三类成分的体内代谢产物，其中1个蒽醌类成分、1个蒽醌类成分代谢产物和1个西红花酸类成分代谢产物活性较强，另1个蒽醌类成分代谢产物活性最强。但以上三类成分剂量较高时也具有一定肝细胞毒性，即具有肝细胞保护和毒性双重作用。

Hepatoprotective ingredients of Yinchenhao Decoction based on the time-spectrum-effect relationship of serum containing drugs

OBJECTIVE To clarify the hepatoprotective material of Yinchenhao decoction (YCHD). METHODS Serum containing drugs (SCDs) of YCHD was obtained from rats intragastrically received YCHD at 10 different time points. Fingerprints of these SCDs were determined by HPLC. The hepatoprotective effects of these SCDs were evaluated in injuried HL-7702 cells. The peaks in fingerprints of SCDs related to pharmacodynamic effects were identified by the analysis of spectrum-effect relationship based on comparison of concentration-time curve and time-effect curve and bivariate correlation analysis. RESULTS Compared by the concentration-time and the time-effect curves, the hepatoprotection of SCDs obviously related to peak 4 to 7, and peak 20 in fingerprints. Peak 8 and 11 to 19 in fingerprints of SCDs had certain protective effects. Peak 1 to 3 and 9 in fingerprints of SCDs were related to hepatotoxicity. According to the bivariate correlation analysis, the hepatoprotection related to peak 1 to 9, 15 to 17, and 20 in fingerprints of SCDs. The effects of peak 4, 5 and 20 were stronger, and peak 6 had the strongest. CONCLUSION The main hepatoprotective ingredients of YCHD derive from anthraquinones from Rhei Radix et Rhizoma, iridoids and crocetins from Gardeniae Fructus, and the metabolites of these three components. The activities of one anthraquinone, one metabolite of anthraquinone and one metabolite of crocetin are stronger, and the activity of another one metabolite of anthraquinone is the strongest. But hepatotoxicity also appears at high doses of these three components.