目的 制备高良姜素-乳清蛋白纳米粒（galangin-whey protein-nanoparticles，Gal-WP-NPs），考察其体内口服药动学行为及其对肝缺血再灌注损伤的保护作用。方法 单因素实验考察各个处方工艺对Gal-WP-NPs的影响。采用Box-Behnken设计-效应面法（Box-Behnken design-response surface methodology，BBD-RSM）得到Gal-WP-NPs最优处方工艺，测定Gal-WP-NPs包封率、载药量、粒径及ζ电位等。冷冻干燥法制备Gal-WP-NPs粉末，透射电子显微镜（transmission electron microscope，TEM）观察Gal-WP-NPs微观形貌，X射线粉末衍射（X-ray powder diffraction，XRPD）法分析Gal-WP-NPs中高良姜素晶型。比较高良姜素及Gal-WP-NPs溶解度，考察Gal-WP-NPs模拟胃肠液中的释药行为。按150 mg/kg（以高良姜素计）ig给予SD大鼠高良姜素和Gal-WP-NPs，计算主要药动学参数及Gal-WP-NPs相对生物利用度。建立SD大鼠肝脏缺血再灌注模型，比较高良姜素和Gal-WP-NPs对肝缺血再灌注损伤的保护作用。结果 Gal-WP-NPs最佳处方工艺：反溶剂相与溶剂相体积比为15.40∶1，乳清蛋白与高良姜素用量比为15.05∶1，超声功率为82.00 W。Gal-WP-NPs包封率、载药量、粒径和ζ电位分别为（94.07±0.98）%、（5.76±0.09）%、（197.39±4.46）nm和（-23.79±0.75）mV。Gal-WP-NPs呈类球形纳米粒，高良姜素在Gal-WP-NPs粉末中以无定形状态存在，溶解度得到极显著提高（P＜0.01），Gal-WP-NPs体外释药行为符合Weibull模型。Gal-WP-NPs达峰时间（t_max）延后至（2.17±0.64）h，半衰期（t_1/2）增加至（5.11±0.95）h，口服相对生物利用度提高至6.58倍。Gal-WP-NPs极大提高了高良姜素对肝缺血再灌注损伤的保护作用。结论 Gal-WP-NPs显著增加了高良姜素口服相对生物利用度，增强了高良姜素对对肝缺血再灌注损伤的保护作用。

Objective To prepare galangin-whey protein-nanoparticles (Gal-WP-NPs), and study its oral pharmacokinetic behavior and protective effect on the hepatic ischemia reperfusion injury in vivo. Methods Single-factor experiments were conducted to investigate the effects of various formulation processes on Gal-WP-NPs. Box-Behnken design-response surface methodology (BBD-RSM) was used to obtain its optimal prescriptions. Entrapment efficiency, drug loading, particle size and ζ potential were determined. Gal-WP-NPs powder was prepared by freeze-drying method. Transmission electron microscopy (TEM) was used to observe the microstructure of Gal-WP-NPs. Galangin crystal in Gal-WP-NPs powder was analyzed by X-ray powder diffraction (XRPD). Solubility of galangin and Gal-WP-NPs powder were compared, and drug release behavior of Gal-WP-NPs in gastrointestinal fluid was determined. SD rats were administered galangin and Gal-WP-NPs via ig at a dose of 150 mg/kg (calculated as galangin), the main pharmacokinetic parameters and relative bioavailability of Gal-WP-NPs were calculated. The model of hepatic ischemia reperfusion injury was established, and then the protective effects of galangin and Gal-WP-NPs were compare on the hepatic ischemia reperfusion injury. Results Optimal formulation processes of Gal-WP-NPs: the volume ratio of the anti-solvent phase to the solvent phase was 15.40:1, the amounts ratio of whey protein to galangin was 15.05:1, and the ultrasonic power was 82.00 W. Envelopment efficiency, drug loading, particle size and ζ potential were (94.07 ± 0.98)%, (5.76 ± 0.09)%, (197.39 ± 4.46) nm and (-23.79 ± 0.75) mV, respectively. Gal-WP-NPs are spherical nanoparticles. Galangin existed as an amorphous state in Gal-WP-NPs powder. Solubility of galangin was significantly increased (P < 0.01), and the release process of Gal-WP-NPs in vitro conformed to the Weibull model. The peak time (t_max) of Gal-WP-NPs was delayed to (2.17 ± 0.64) h, half-life period (t_1/2) was increased to (5.11 ± 0.95) h, and oral relative bioavailability was increased to 6.58 times. Gal-WP-NPs significantly enhanced the protective effect of galangin on the hepatic ischemia reperfusion injury. Conclusion Gal-WP-NPs significantly increased the oral relative bioavailability of galangin and enhanced the protective effect on the hepatic ischemia reperfusion injury.

R283.6

河南省科技厅科技攻关项目（232102311023）；2023年河南省高等教育教学改革研究与实践项目（2023SJGLX125Y）