目的 制备三叶苷-磷脂复合物/壳聚糖纳米粒（trifolin-phospholipids complex/chitosan nanoparticles，Tri-PC/CS-NPs），考察Tri-PC/CS-NPs体内药动学行为及对急性肝损伤的保护作用。方法 采用溶剂挥发法制备三叶苷-磷脂复合物（trifolin-phospholipids complex，Tri-PC）。采用单因素实验结合Box-Behnken设计-效应面法（Box-Behnken design-response surface methodology，BBD-RSM）优化Tri-PC/CS-NPs处方工艺。采用透射电子显微镜（transmission electron microscope，TEM）观察Tri-PC/CS-NPs形貌，X射线粉末衍射法（X-ray powder diffraction，XRPD）分析Tri-PC/CS-NPs晶型。比较三叶苷、Tri-PC、Tri-PC/CS-NPs的饱和溶解度、油水分配系数及其在模拟胃肠液中的释药行为。SD大鼠分别ig给予三叶苷、Tri-PC、Tri-PC/CS-NPs，测定血药浓度，计算主要药动学参数。建立急性肝损伤大鼠模型，比较三叶苷、Tri-PC、Tri-PC/CS-NPs对急性肝损伤大鼠的保护作用。结果 Tri-PC/CS-NPs最佳处方：水相体积为20.50 mL，Tri-PC与壳聚糖质量比为4.15∶1，泊洛沙姆188质量分数为0.10%。Tri-PC/CS-NPs的包封率、载药量、粒径及ζ电位分别为（84.80±1.21）%、（15.56±0.10）%、（197.74±5.58）nm和（32.69±1.14）mV。Tri-PC/CS-NPs外貌为类球形。三叶苷在Tri-PC和Tri-PC/CS-NPs中均以无定形形态存在。Tri-PC和Tri-PC/CS-NPs极显著性提高了三叶苷的饱和溶解度及油水分配系数（P＜0.01）。Tri-PC和Tri-PC/CS-NPs在18 h内累积释放率分别为56.68%和88.45%。Tri-PC/CS-NPs的释药行为符合Weibull模型。以三叶苷为参考，Tri-PC和Tri-PC/CS-NPs相对生物利用度分别增加至2.41倍和5.36倍。Tri-PC和Tri-PC/CS-NPs有效增强了三叶苷对急性肝损伤大鼠的保护作用，且Tri-PC/CS-NPs效果优于Tri-PC。结论 Tri-PC/CS-NPs改变了三叶苷的体内药动学行为，有效促进了三叶苷的体内吸收，并增强了三叶苷对急性肝损伤的保护作用。

Objective To prepare trifolin-phospholipids complex/chitosan nanoparticles (Tri-PC/CS-NPs), and investigate its in vivo pharmacokinetic behavior and protective effect on acute liver injury. Methods Trifolin-phospholipids complex (Tri-PC) was prepared by solvent evaporation method. Single factor experiments combined Box-Behnken design-response surface methodology (BBD-RSM) were used to optimize the prescriptions of Tri-PC/CS-NPs. Transmission electron microscope (TEM) was employed to observe microscopic appearance of Tri-PC/CS-NPs, X-ray powder diffraction (XRPD) was used to analyze crystal form of Tri-PC/CS-NPs. Saturated solubility, oil/water partition coefficient and drug release behavior in simulated gastrointestinal fluids of trifolin, Tri-PC and Tri-PC/CS-NPs were compared. SD rats were administered intragastrically with trifolin, Tri-PC and Tri-PC/CS-NPs, respectively. Blood drug concentration was determined, and the main pharmacokinetic parameters were calculated. The model of acute liver injury was established, and the protective effect of trifolin, Tri-PC, Tri-PC/CS-NPs on acute liver injury were compared. Results Optimal formulations of Tri-PC/CS-NPs: the volume of water phase was 20.50 mL, the mass ratio of Tri-PC to chitosan was 4.15:1, and the mass fraction of Poloxamer 188 was 0.10%. Envelopment efficiency, drug loading, particle size and ζ potential were (84.80 ± 1.21)%, (15.56 ± 0.10)%, (197.74 ± 5.58) nm and (32.69 ± 1.14) mV, respectively. The appearance of Tri-PC/CS-NPs were spherical. The state of trifolin changed into amorphous form in Tri-PC and Tri-PC/CS-NPs. Tri-PC and Tri-PC/CS-NPs significantly enhanced the saturated solubility and oil/water partition coefficient of trifolin (P < 0.01). Cumulative release rate of Tri-PC and Tri-PC/CS-NPs were 56.68% and 88.45% in 18 h, respectively. The drug release process of Tri-PC/CS-NPs conformed to the Weibull model. Compared with trifolin, the relative bioavailability of Tri-PC and Tri-PC/CS-NPs was enhanced to 2.41-fold and 5.36-fold, respectively. Tri-PC and Tri-PC/CS-NPs effectively enhanced the protective effect of trifolin against acute liver injury, and the protective effect of Tri-PC/CS-NPs was superior to that of Tri-PC. Conclusion Tri-PC/CS-NPs changed the pharmacokinetic behavior of trifolin in vivo, effectively promoted the absorption of trifolin, and enhanced the protective effect of trifolin on acute liver injury.

R283.6

国家自然科学基金项目（81670088）；2025年度郑州市医疗卫生领域科技创新指导计划项目（2025-421）