目的 根据历代医籍和本草记载的炮制方法，对比补骨脂Psoraleae Fructus不同炮制品水提物的急性毒性和肝毒性差异，为补骨脂的炮制方法使用提供参考。方法 采用超高效液相色谱-串联三重四极杆质谱和标准曲线法对补骨脂生品和补骨脂不同炮制品水提物进行补骨脂素和异补骨脂素的含量测定，并以AB野生型斑马鱼、Tg（lfabp: EGFP）肝脏细胞绿色荧光标记转基因斑马鱼和Tg（lysc: DsRed2）中性粒细胞红色荧光标记转基因斑马鱼为实验动物模型，给予补骨脂生品和补骨脂不同炮制品的水提物通过观察补骨脂不同炮制品对斑马鱼的半数致死浓度（median lethal concentration，LC₅₀）、最小致死浓度（minimum lethal concentration，LC₁）、斑马鱼形态变化、畸形评分等的影响，比较补骨脂不同炮制品的急性毒性差异。通过观察不同给药组对斑马鱼肝脏荧光强度、肝脏面积和体积、肝脏病理切片情况、肝脏区域中性粒细胞数量的影响，以及检测炎症相关基因的mRNA水平变化情况，综合评价补骨脂不同炮制品的肝毒性差异。结果 补骨脂不同炮制品水提物中补骨脂素和异补骨脂素含量存在差异，补骨脂生品中补骨脂素和异补骨脂素含量最高，雷公法制品中异补骨脂素含量最低，清炒法制品中补骨脂素含量最低。补骨脂不同炮制品水提物急性毒性存在差异，补骨脂生品水提物会导致斑马鱼出现严重的急性毒性，表现为出现心包水肿、脊柱弯曲、鱼鳔消失、卵黄囊吸收延滞，而不同炮制品中补骨脂雷公法制品的急性毒性减毒效果最优，补骨脂酒炙法制品、补骨脂盐炙法制品和补骨脂清炒法制品急性毒性的减毒效果依次减弱。补骨脂生品水提物会导致斑马鱼出现严重的肝脏毒性，包括肝脏结构缺失、肝脏荧光强度减弱、肝脏荧光面积和体积减少、肝脏细胞间出现空泡化，并导致肝脏区域的中性粒细胞聚集，上调炎症相关因子表达水平。基于斑马鱼模型不同炮制法中补骨脂雷公法的肝脏毒性减毒效果最优，补骨脂酒炙法和补骨脂清炒法肝毒性的减毒效果依次减弱，而补骨脂盐炙法则增强肝脏毒性毒效果。结论 不同炮制法对补骨脂急性毒性均有减毒作用，这可能与补骨脂素和异补骨脂素含量的减少有关，对肝毒性的影响存在差异。基于斑马鱼模型雷公法炮制对急性毒性和肝毒性的减毒效果最佳，且通过减少肝脏区域炎症发生和中性粒细胞的聚集减轻了补骨脂的肝毒性，而盐炙法则增加肝脏区域炎症发生和中性粒细胞聚集加重肝毒性。

Objective To compare the acute toxicity and hepatotoxicity of water extracts from different processed products of Buguzhi (Psoraleae Fructus) according to the processing methods recorded in medical books and materia medica in past dynasties, and provide the reference for the use of processing methods of Psoraleae Fructus. Methods The contents of psoralen and isopsoralen in water extracts of raw and processed Psoraleae Fructus were determined by ultra performance liquid chromatography-tandem triple quadrupole mass spectrometry and standard curve method. Wild-type AB zebrafish, transgenic zebrafish Tg (lfabp: EGFP) with green fluorescent labeling of liver cells and transgenic zebrafish Tg (lysc: DsRed2) with red fluorescent labeling of neutrophils were taken as experimental research model. The water extracts of raw and processed Psoraleae Fructus were given. By observing the effects of different processed Psoraleae Fructus on the median lethal concentration (LC₅₀) and minimum lethal concentration (LC₁), morphological changes and deformity scores of zebrafish, the acute toxicity differences of different processed Psoraleae Fructus were compared. By observing the effects of different drug groups on the fluorescence intensity, liver area and volume, liver pathological sections and the number of neutrophils in the liver area of zebrafish, and detecting the changes in mRNA levels of inflammation-related genes, the hepatotoxicity differences of different processed products of Psoraleae Fructus were comprehensively evaluated. Results The contents of psoralen and isopsoralen in different processed products of Psoraleae Fructus were different. The content of psoralen and isopsoralen in raw products of Psoraleae Fructus was the highest, the content of isopsoralen in Leigong products was the lowest, and the content of psoralen in stir-fried products was the lowest. The acute toxicity of water extracts from different processed products of Psoraleae Fructus was different. The water extracts from raw products of Psoraleae Fructus leaded to serious acute toxicity of zebrafish, such as pericardial edema, spinal curvature, the disappearance of the swim bladder and delayed absorption of the yolk sac. Among the products processed by different methods, the acute toxicity attenuation effect of Psoraleae Fructus by the Leigong method was the best, and the acute toxicity attenuation effects of Psoraleae Fructus by wine, Psoraleae Fructus by salt and Psoraleae Fructus by stir-frying method were weakened in turn. The water extract of Psoraleae Fructus could cause serious hepatotoxicity in zebrafish, including loss of liver structure, weakening of liver fluorescence intensity, reduction of liver fluorescence area and volume, vacuolation between liver cells, and aggregation of neutrophils in the liver area, which could promote the expression level of inflammation-related factors. Among the different processing methods, the Leigong method of Psoraleae Fructus had the best attenuation effect on liver toxicity, while the methods of frying Psoraleae Fructus with wine and frying Psoraleae Fructus had weakened the attenuation effect on liver toxicity in turn, while the method of roasting Psoraleae Fructus with salt had enhanced the attenuation effect on liver toxicity. Conclusion Different processing methods have attenuated the acute toxicity of Psoraleae Fructus, this may be related to the decrease of psoralen and isopsoralen content, but their effects on liver toxicity are different. The Leigong method has the best attenuation effect on acute toxicity and hepatotoxicity, and reduces the hepatotoxicity of Psoraleae Fructus by reducing inflammation and neutrophil aggregation in liver area, while the salt roasting method increases inflammation and neutrophil aggregation in liver area and aggravates hepatotoxicity.

国家自然科学地区科学基金项目（82060782）；国家自然科学地区科学基金项目（82160821）；天山英才项目（2022TSYCLJ009，2022TSYCCX0021）；自治区重点研发项目（2022B03012-4）；中医药传承与创新“百千万”人才工程（岐黄工程）-青年岐黄学者，国家岐黄学者；2020年新疆维吾尔自治区高层次人才引进工程（柔性引进人才）项目；自治区自然科学基金资助项目（2022D01B191）