目的 制备共载紫草素和藁本内酯的脂质体（liposomes co-loaded with shikonin and ligustilide，Lip@Shi/Lig）并优化其制备工艺，对其进行相关表征及体外透皮性能研究。方法 应用CCK-8法和Synergy Finder分析工具，以细胞活力为评价指标，确定紫草素与藁本内酯的最佳联用比例；采用薄膜分散法制备Lip@Shi/Lig，以包封率为评价指标，通过单因素实验筛选Lip@Shi/Lig的最佳处方；通过外观与形貌观察，粒径、多分散指数（polydispersity index，PDI）、ζ电位测定，X射线衍射（X-ray diffraction，XRD）分析和傅里叶变换红外光谱法（Fourier transform infrared spectroscopy，FT-IR）分析等表征手段，对Lip@Shi/Lig进行质量评价；最后，采用Franz扩散池法考察Lip@Shi/Lig的经皮渗透能力及真皮滞留性能。结果 紫草素与藁本内酯的最佳联用比例为1∶1。Lip@Shi/Lig的最佳制备工艺为蛋黄卵磷脂质量浓度10 mg/mL，蛋黄卵磷脂与胆固醇质量比4∶1，蛋黄卵磷脂与总药物质量比1∶15，超声时间5 min；该方法制备的Lip@Shi/Lig形态规整，为类圆形囊泡结构，紫草素与藁本内酯的包封率分别为（98.16±0.67）%、（97.20±0.76）%，Lip@Shi/Lig粒径为（88.62±0.26）nm，PDI为0.246±0.013，ζ电位为（-36.57±1.65）mV。XRD与FT-IR结果表明，紫草素和藁本内酯被成功包裹于脂质体中；Lip@Shi/Lig中紫草素与藁本内酯在30 h内的累积透皮量分别为（82.97±0.72）、（81.57±3.59）μg/cm²，真皮滞留量（Q_s）分别为（6.12±0.18）、（8.08±0.04）μg/cm²，相比单体药物和单载药脂质体均有所提高。结论 成功制备了具有良好透皮性能和稳定性的Lip@Shi/Lig，并显著改善了紫草素与藁本内酯的透皮性能和Q_s，为其进一步体内研究和未来临床应用提供了实验依据。

Objective To prepare liposomes co-loaded with shikonin and ligustilide (Lip@Shi/Lig), optimize its preparation process, and to investigate its characterization and transdermal performance in vitro. Methods The CCK-8 assay and Synergy Finder analysis tool were used to determine the optimal combination ratio of shikonin and ligustilide based on cell viability. Lip@Shi/Lig was prepared using the thin-film dispersion method, and the optimal formulation of Lip@Shi/Lig was screened through single-factor investigation using encapsulation efficiency as the evaluation criterion. The quality evaluation of Lip@Shi/Lig was conducted using characterization methods, including appearance and morphology observation, particle size, polydispersity index (PDI), ζ potential determination, X-ray diffraction (XRD) analysis, and Fourier transform infrared spectroscopy (FT-IR) analysis. Finally, the transdermal permeability and dermal retention performance of Lip@Shi/Lig were investigated using the Franz diffusion cell method. Results The optimal combined ratio of shikonin and ligustilide was 1∶1. The optimal conditions were determined as follows: egg yolk lecithin concentration of 10 mg/mL, phospholipid-cholesterol ratio of 4∶1, total drug-phospholipid ratio of 1∶15, and ultrasonication time of 5 min. The resulting Lip@Shi/Lig exhibited regular, spherical vesicle morphology, with encapsulation efficiencies of shikonin and ligustilide at (98.16 ±0.67)% and (97.20 ±0.76)%, respectively, particle size of (88.62 ±0.26) nm, PDI of 0.246 ±0.013, and ζ potential of (-36.57 ±1.65) mV. XRD and FT-IR results indicated that shikonin and ligustilide were successfully encapsulated in the liposomes. The cumulative penetration of shikonin and ligustilide in Lip@Shi/Lig within 30 h were (82.97 ±0.72) μg/cm² and (81.57 ±3.59) μg/cm², respectively, with dermal retention rates of (6.12 ±0.18) μg/cm² and (8.08 ±0.04) μg/cm², respectively, both of which were significantly higher than those of the single drug and the single-drug-loaded liposomes. Conclusion Lip@Shi/Lig was successfully prepared with favorable transdermal properties and stability. It significantly enhanced the transdermal penetration and dermal retention of both shikonin and ligustilide, providing a solid experimental foundation for further in vivo studies and potential clinical applications.

R283.6

国家自然科学基金资助项目(82173982);广东省自然科学基金资助项目(2022A1515011382)