目的 制备壳聚糖修饰的大黄酸脂质体（ CS-Rhe-Lips） 凝胶，评价其制剂学特性、透皮递送效率及抗皮肤光老化活性。方法 以包封率、载药量及粒径为关键评价指标，采用单因素结合 Box-Behnken 响应面法优化 CS-Rhe-Lips 的制备处方；通过透射电子显微镜（ TEM）观察其微观形态。以卡波姆 940 为凝胶基质，将优化后的 CS-Rhe-Lips 混悬液制备为 CSRhe-Lips 凝胶，考察其体外释药行为及流变学特征；采用 Franz 扩散池法评价该凝胶的经皮渗透性能及皮肤滞留量。建立紫外线照射诱导的昆明小鼠皮肤光老化模型， 将小鼠按体质量随机分为对照组、 模型组、 维生素 C（ 阳性药， 10 mg·kg^-1） 组、大黄酸凝胶（ 20 mg·kg^-1） 组、 大黄酸脂质体（ Rhe-Lips， 20 mg·kg^-1） 凝胶组及 CS-Rhe-Lips 凝胶低、 高剂量组（ 10、 20 mg·kg^-1）组。通过皮损面积和严重程度（ PASI） 评分评价皮损面积及严重程度，检测皮肤组织炎症因子水平与皮肤含水量，结合苏木素-伊红（ HE） 染色、 Masson 染色观察皮肤病理形态及胶原纤维变化。结果 确定 CS-Rhe-Lips 的最佳制备处方为：药物与磷脂用量比 9.88∶ 1.00、磷脂与胆固醇用量比 5.18∶ 1.00、壳聚糖质量分数 0.19%。优化后 CS-Rhe-Lips 的平均包封率为（ 85.65±0.85） %，载药量为（ 3.93±0.09） %，粒径为（ 185.83±7.55） nm， Zeta 电位为（ 25.15±1.09） mV。TEM 观察显示其形态圆整，呈类圆形或圆形。CS-Rhe-Lips 凝胶体外 24 h 累积释放度达 76.73%，具有良好的固体弹性特征，倒置后无流淌现象。与大黄酸凝胶相比， CS-Rhe-Lips 凝胶的大黄酸透皮速率、累积渗透量及皮肤滞留量分别提高 2.90 倍、 2.87 倍和 3.28 倍。动物实验结果显示， CS-Rhe-Lips 凝胶能显著降低光老化小鼠皮肤 PASI 评分，抑制炎症因子的异常升高，提高皮肤含水量，改善光老化皮肤的外观形态及组织结构，增加胶原纤维含量；其抗光老化作用显著优于大黄酸凝胶及 Rhe-Lips凝胶，且高剂量组效果更优。结论 CS-Rhe-Lips 凝胶具有理想的缓释特性与流变学性能，可显著改善大黄酸的透皮递送效率，增强其抗皮肤光老化活性。

Objective To prepare chitosan-coated rhein (CS-Rhe-Lips) liposomes, its pharmaceutical characteristics, transdermal delivery efficiency and anti-skin photoaging effect was investigated. Methods Envelopment efficiency, drug loading and particle size were used as key evaluation indicators, single factor study and Box-Behnken design-response surface method (BBD-RSM) were employed to optimize the prescriptions of CS-Rhe-Lips. Transmission electron microscope (TEM) was employed to observe the microscopic morphology of CS-Rhe-Lips. CS-Rhe-Lips gel was formulated using carbopol 940 as gel matrix after optimized the prescriptions of CS-Rhe-Lips suspension, its drug release behavior in vitro and rheological characteristics were also investigated. Franz diffusion cell method was used to investigate the transdermal properties and skin retention of CS-Rhe-Lips gel. Ultraviolet irradiation-induced skin photoaging model of KM mice was established, and then randomly divided into control group, model group, vitamin C group (positive drug, 10 mg·kg-1), rhein gel group (Rhe-Lips, 20 mg·kg-1), CS-Rhe-Lips gel low-dose and high-dose group (10, 20 mg·kg-1). The area and severity of skin lesions were evaluated by the PASI score. The levels of inflammatory factors in skin tissue and the skin water content were detected. The pathological morphology and changes of collagen fibers in the skin were observed through hematoxylin-eosin (HE) staining and Masson staining. Results The optimal formulation of CS-Rhe-Lips: carriers to drug ratio was 9.88∶ 1.00, phospholipids to cholesterol ratio was 5.18∶ 1.00, and chitosan concentration was 0.19%. Envelopment efficiency, drug loading, particle size and Zeta potential of CS-Rhe-Lips were (85.65 ±0.85) %, (3.93 ±0.09) %, (185.83 ±7.55) nm, (25.15 ±1.09) mV, respectively. TEM results showed that the microscopic morphology of CS-Rhe-Lips were regular, presenting as an oval or circular shape. The cumulative release rate of CS-Rhe-Lips gel was increased to 76.73% in 24 h, which had solid properties and did not flow after inversion. Compared with Rhe gel, CS-Rhe-Lips gel enhanced the transdermal rate, cumulative permeation amount, and skin retention of rhein by 2.90, 2.87, and 3.28-fold, respectively. The results of animal experiments showed that CS-RheLips gel significantly reduced the PASI, suppressed the increase of inflammatory factors in photoaging skin tissue, increased skin moisture, improved the appearance and tissue structure of photoaging skin, and increased collagen fibers. The anti-skin photoaging effect of CS-Rhe-Lips gel was superior than that of rhein gel and Rhe-Lips gel, and the effect of high-dose group of CS-Rhe-Lips gel was better. Conclusion CS-Rhe-Lips gel exhibited ideal sustained release characteristics and rheological properties, significantly improving the transdermal delivery efficiency of rhein, and enhancing anti-skin photoaging activity.

R283;R944.9;R758.1

2024 年度河南省科技攻关项目（242102110111）