目的 探讨雷公炮制过程补骨脂核心成分香豆素类、黄酮类和补骨脂酚质量传递及对毒/效的影响。方法 采用HPLC法检测补骨脂雷公炮制过程（酒浸、水浸、蒸制）核心成分香豆素类、黄酮类、补骨脂酚的动态变化。采用受精后1～6 d（days post fertilization，dpf）的斑马鱼评价补骨脂雷公炮制前后的安全性，观察鱼脏器形态、记录死亡数并计算斑马鱼半数死亡浓度（half lethal concentration，LC₅₀）；采用25 μmol/L泼尼松龙诱导斑马鱼骨丢失模型，茜素红对培养至8 dpf的各给药组斑马鱼幼鱼骨骼染色，进行显微观察、数码成像，并用图像软件定量分析骨骼染色区域评价各给药组的抗骨质疏松活性。结果 白酒浸液主要含脂溶性成分如（异）补骨脂素、黄酮类和补骨脂酚，浸出率为20.91%～63.93%，而黄酒浸液与水浸液相似，仅含少量（异）补骨脂苷，（异）补骨脂素或新补骨脂异黄酮（浸出率为0.22%～6.15%），进一步蒸制后，（异）补骨脂苷含量明显降低至生品的9.54%～13.71%，而（异）补骨脂素含量则升高至生品的134.27%～143.13%，提示苷向苷元转化；此外发现4′-O-甲基补骨脂查耳酮含量明显升高（为蒸制前的247.09%～288.18%），推测可能由其他黄酮经加热转化而来。斑马鱼评价结果表明，白酒雷公炮制品安全性优于黄酒雷公炮制品，且均优于生品：炮制后，致鱼畸变浓度升高，致鱼最早死亡时间推迟2～3 d，鱼最大死亡率减少45%～70%，LC₅₀值升高3～4倍；雷公炮制品和生品均可增加斑马鱼头骨矿化面积和吸光度值。结论 雷公炮制过程（酒浸、水浸及蒸制）中补骨脂核心成分发生物理浸出、苷向苷元生物转化以及黄酮甲基化化学修饰等不同形式动态变化，雷公炮制具有减毒存效作用，新补骨脂异黄酮、补骨脂甲素和补骨脂酚含量降低是补骨脂炮制减毒的关键因素。

Objective To investigate quality delivery of core components such as coumarins, flavonoids, and bakuchiol of Buguzhi (Psoraleae Fructus) during Leigong processing and the influence on toxicity/efficacy. Methods HPLC method was used to detect the dynamic changes of coumarins, flavonoids, and bakuchiol, the core components of Psoraleae Fructus in the Leigong processing process (wine soaking, water soaking, and steaming). The safety of Psoraleae Fructus before and after Leigong processing was evaluated in zebrafish embryos/larvae (1—6 dpf, days post fertilization), organ morphology was observed, mortality was recorded, and the half lethal concentration (LC₅₀) was calculated. The zebrafish osteoporosis model was induced with 25 μmol/L prednisolone; microscopic observation and digital imaging of zebrafish larvae of each treatment group cultured to 8 dpf were performed using alizarin red, and the bone staining area was quantitatively analyzed by image software to evaluate the anti-osteoporosis activity of the above samples. Results Chinese liquor infusion mainly contained lipophilic components such as (iso) psoralen, flavonoids, and bakuchiol, and the leaching rate was 20.91%—63.93%, while the rice wine infusion was similar to that of water infusion, which only contained a small amount of hydrophilic (iso) psoralenoside and lipophilic (iso) psoralen or neobavaisoflavone (leaching rate was 0.22%—6.15%), and then after further steaming, the content of (iso) psoralenoside was significantly reduced to 9.54%—13.71% of the raw products, while the content of (iso) psoralen increased to about 134.27%—143.13% of the raw products, indicating that glycoside was converted to aglycone. In addition, the content of 4′-O-methylbroussochalcone was significantly increased (about 247.09%—288.18% of the products before steaming), which may be converted from other flavonoids by heating. The results of zebrafish evaluation showed that the safety of Leigong products using rice wine was better than that of using Chinese liquor, and both were better than that of raw products; After processing, the concentration of fish distortion was increased, the earliest death time of fish was delayed by 2—3 d, the maximum mortality rate of fish was reduced by 45%—70%, and the LC₅₀ value of fish increased by 3—4 times. Both Leigong products and raw products could increase the mineralized area and cumulative optical density value of zebrafish skulls. Conclusion Leigong processing (wine soaking, water soaking, and steaming) causes different dynamic changes of core components of Psoraleae Fructus, such as physical leaching of core components, bioconversion of glycoside to aglycone, and chemical modification of flavonoid methylation, and the evaluation of zebrafish shows that the Leigong processing method can reduce toxicity and save effectivity, and the content reduction of neobavaisoflavone, bavachin, and backuchiol is the key factor of toxicity attenuation.

R285.5

“重大新药创制”科技重大专项资助项目(2017ZX09301056); 江苏省中医药科技发展计划项目(ZD202225); 江苏省中医药研究院高质量研究成果后补助项目(HBZ202501)