目的 制备柠檬苦素负载玉米醇溶蛋白/岩藻多糖纳米粒（Lim-Zein/Fuc-NPs），考察其体内药动学特征，并评价其对高尿酸血症大鼠的肾脏保护作用。方法 采用单因素实验结合Box-Behnken设计-效应面法优化Lim-Zein/Fuc-NPs处方工艺；通过透射电镜（TEM）观察Lim-Zein/Fuc-NPs微观形貌，采用X-射线粉末衍射法（XRPD）分析柠檬苦素（Lim）在LimZein/Fuc-NPs中的晶型状态。测定Lim-Zein/Fuc-NPs饱和溶解度，采用透析法比较Lim和Lim-Zein/Fuc-NPs在pH 2.0、6.8磷酸盐缓冲溶液中的体外释药行为。SD大鼠分别ig给予Lim和Lim-Zein/Fuc-NPs，计算主要药动学参数及相对口服生物利用度。构建高尿酸血症大鼠模型，比较Lim和Lim-Zein/Fuc-NPs对尿酸（UA）、尿素氮（BUN）和血肌酐（CRE）水平的影响，通过苏木精-伊红（HE）染色观察大鼠肾脏病理学变化。结果 Lim-Zein/Fuc-NPs最佳处方工艺为：玉米醇溶蛋白与Lim用量比14.74∶ 1.00，岩藻多糖与玉米醇溶蛋白用量比1.99∶ 1.00，制备时间1.25 h。该工艺制备的Lim-Zein/Fuc-NPs包封率、载药量、粒径及Zeta电位分别为（84.31±0.96）%、（1.93±0.06）%、（185.77±4.69） nm、（-32.77±1.09） mV。LimZein/Fuc-NPs呈类球形，无黏连现象。XRPD结果显示，Lim被包载后由晶态转变为无定形态，饱和溶解度和18 h累积释放率较游离药物均大幅提升。体内药动学结果表明，与Lim比较，Lim-Zein/Fuc-NPs可显著改变药物体内吸收行为，相对口服生物利用度提升至7.25倍。体内药效结果显示，Lim-Zein/Fuc-NPs可极显著降低高尿酸血症大鼠血清UA、BUN、CRE水平（P＜0.01）。HE染色结果证实Lim-Zein/Fuc-NPs能有效改善肾脏病理损伤。结论 Lim-Zein/Fuc-NPs显著增加了Lim的溶解度、释放率及生物利用度，进一步增强其对高尿酸血症大鼠的肾脏保护效果，具备临床开发价值。

Objective To prepare limonin-loaded by zein-fucoidan nanoparticles (Lim-Zein/Fuc-NPs), and investigate its pharmacokinetics and protective effect on hyperuricemic rats in vivo. Methods Main influencing factors of Lim-Zein/Fuc-NPs were investigated by single factor experiments, Box-Behnken response surface design method was employed to optimize prescriptions of Lim-Zein/Fuc-NPs, and Lim-Zein/Fuc-NPs powder was prepared. Transmission electron microscope (TEM) was used to observe the microscopic appearance of Lim-Zein/Fuc-NPs, X-ray powder diffraction (XRPD) was employed to analyze the crystal form of Lim in Lim-Zein/Fuc-NPs powder. Saturated solubility and cumulative release rate of Lim-Zein/Fuc-NPs were determined. SD rats were administered intragastrically with Lim and Lim-Zein/Fuc-NPs respectively, their pharmacokinetic parameters and relative oral bioavailability were calculated. Hyperuricemic model was establish, and the effects of Lim and Lim-Zein/Fuc-NPs on uric acid (UA), blood urea nitrogen (BUN), and creatinine (CRE) levels were compared. Hematoxylin and eosin (HE) staining was used to observe pathological changes in kidney tissue. Results Optimal formulations of Lim-Zein/Fuc-NPs: the amounts ratio of zein to limonin is 14.74∶ 1.00, the amounts ratio of fucoidan to zein is 1.99∶ 1.00, and the preparation time was 1.25 h. Envelopment efficiency, drug loading, particle size and Zeta potential were (84.31±0.96) %, (1.93±0.06) %, (185.77±4.69) nm and (–32.77±1.09) mV, respectively. The appearance of Lim-Zein/Fuc-NPs were spheroidal with no adhesion phenomenon. The state of Lim changed into amorphous form in Lim-Zein/Fuc-NPs powder. Saturated solubility and cumulative release rate in 18 h were increased greatly. Compared with Lim, the in vivo pharmacokinetic behavior of Lim-Zein/Fuc-NPs changed significantly, and the relative oral bioavailability was increased to 7.25 times. The levels of UA, BUN, and CRE in Lim-Zein/Fuc-NPs group significantly decreased compared to Lim group (P < 0.01). HE staining confirmed that Lim-Zein/Fuc-NPs effectively improved the pathological changes of kidney tissue. Conclusion Lim-Zein/Fuc-NPs significantly increased the solubility, drug release rate and oral bioavailability of Lim, and enhanced the protective effect of Lim on hyperuricemic rats and possessing clinical development value.

R285.5

国家自然科学基金资助项目（31371386；30871239）