目的 基于斑马鱼模型结合相关分析高效辨识补骨脂毒性成分群。方法 采用高效液相色谱法测定7个产地18个批次补骨脂中4个香豆素类、4个黄酮类和补骨脂酚的含量。以斑马鱼（6 dpf）死亡率、脏器中毒形态、半数致死浓度（lethal concentration 50，LC₅₀）值为指标，采用斑马鱼模型评价18个批次补骨脂水煎液的毒性。将各类成分的含量分别与斑马鱼LC₅₀值进行相关分析，辨识毒性成分群，并通过对代表成分的斑马鱼毒性评价进行验证。结果 不同产地18个批次补骨脂水煎液中香豆素类、黄酮类、补骨脂酚的含量差异较大，4个香豆素类成分、黄酮成分（新补骨脂异黄酮）和补骨脂酚含量最大值分别是最小值的3.47～5.60、5.30、6.47倍。斑马鱼毒性评价结果显示18个批次补骨脂水煎液对斑马鱼毒性差异较大，其中LC₅₀最大值是最小值的7.2倍。毒性与成分含量相关分析结果表明，斑马鱼毒性与黄酮类成分、补骨脂酚的相关性较好（R²分别为0.623 4、0.587 8），与香豆素类成分的相关性较低（R²为0.187 7）。各类代表成分的毒性评价结果表明，补骨脂酚和3个黄酮成分（补骨脂甲素、新补骨脂异黄酮、补骨脂乙素）是补骨脂致毒的关键成分，香豆素类成分的安全性相对较好。结论 斑马鱼模型可高效、灵敏反映不同批次补骨脂毒性的差异，通过毒性与成分含量的相关分析，能够快速发现补骨脂中毒性相关成分群，为从中药复杂体系快速发现毒性及其相关物质提供依据。

Objective To efficiently identify the toxic component groups of Psoraleae Fructus (PF) based on zebrafish model combined with correlation analysis. Methods Contents of four coumarins, four flavonoids and bakuchiol in 18 batches of PF from seven producing areas were determined by HPLC. Zebrafish model was used to evaluate the toxicity of 18 batches of PF decoction, with mortality, organ poisoning form, and LC₅₀ value of zebrafish (6 dpf) as indicators. Correlation analysis between contents of various components and LC₅₀ value of zebrafish (6 dpf) were carried out to identify toxicity related components, and the zebrafish toxicity evaluation results of the representative components were verified. Results Contents of coumarins, flavonoids and bakuchiol in 18 batches of PF decoction from different producing areas were quite different. The maximum content of four coumarins, flavonoid (neobavaisoflavone) and bakuchiol were respectively 3.47-5.60, 5.30 and 6.47 times of the minimum. The zebrafish toxicity evaluation results showed that the toxicity of 18 batches of PF decoction was quite different, and LC₅₀ (6 dpf) of the maximum was 7.2 times of the minimum. The results of correlation analysis between toxicity and component content showed that zebrafish toxicity had a good correlation with flavonoids and bakuchiol (R² were respectively 0.623 4 and 0.587 8), while the correlation with coumarins was relatively low (R² was 0.187 7). Toxicity evaluation results of various representative components showed that bakuchiol and three flavonoids (bavachin, neobavaisoflavone, isobavachalcone) were the key toxic components of PF, while coumarins was relatively safe. Conclusion Zebrafish model could efficiently and sensitively reflect toxicity differences of different batches of PF, of which toxicity relevant component groups could be efficiently identified through correlation analysis between toxicity and content of components, providing a reference for quickly discovering toxicity and related substances from the complex system of traditional Chinese medicine.

R285.5;R285.53

“重大新药创制”科技重大专项资助项目（2017ZX09301-056）；国家自然科学基金资助项目（81573833）；江苏省医学创新团队项目（CXTDB2017003）