目的 比较乙醇、丙二醇、混合醇（乙醇-丙二醇2：8）3种柔软剂对制备光甘草定醇质体（GLA-ES）的影响，为难溶性药物的醇质体研究提供柔软剂选择依据。方法 分别选用乙醇、丙二醇、混合醇（乙醇-丙二醇2：8）作为柔软剂，采用注入-超声结合法制备GLA-ES，并进行不同GLA-ES的形态、粒径、Zeta电位、包封率、稳定性和体外释药等考察。以黑色素瘤B16-OVA细胞内酪氨酸酶活性评价GLA-ES抑制黑色素合成能力，采用铁氰化钾还原力实验评估其抗氧化效果，并使用人表皮HaCaT细胞和大鼠皮肤进行初步安全性评价。结果 所得GLA-ES为黄色半透明液体，具有囊泡状磷脂双分子层结构；光甘草定乙醇醇质体（GLA-Et-ES）、丙二醇醇质体（GLA-PG-ES）和混合醇醇质体（GLA-MA-ES）的平均粒径分别为（34.24±0.29）、（62.31±1.66）、（41.20±1.13）nm，电位分别为（-41.0±1.8）、（-32.9±0.2）、（-35.8±1.6）mV，包封率分别为（91.47±2.39）%、（87.33±1.31）%、（91.39±3.59）%，在4℃放置20 d较稳定；其体外释药行为均符合Higuchi方程，具有一定缓释效果。与光甘草定混悬液相比，GLA-Et-ES、GLA-PG-ES和GLA-MA-ES对黑色素瘤B16-OVA细胞的酪氨酸酶活性抑制率分别提高了38.07%、19.58%、40.42%；铁氰化钾还原力实验结果也显示GLA-ES有较强的体外抗氧化作用；GLA-ES对正常细胞几乎没有毒性，对大鼠皮肤无刺激性。结论 3种柔软剂均可制备GLA-ES，其抑制黑色素生成、抗氧化作用增强，安全性良好，为进一步开发美白护肤产品或医药外用制剂奠定基础。对于光甘草定这类难溶性药物，选择混合醇（乙醇-丙二醇2：8）作为柔软剂，所制备的醇质体的包封率、稳定性优于单用乙醇或丙二醇。

Objective To compare the effects of softeners including ethanol, propylene glycol and mixed alcohol (ethanol-propylene glycol 2:8) on the preparation of glabridin ethosomes (GLA-ES), and provide the selection basis of the softeners for studying the ethosomes of insoluble drugs. Methods GLA-ES were prepared by injection-ultrasonic binding method with ethanol, propylene glycol and mixed alcohol (ethanol-propylene glycol, 2:8) as softeners. The morphology, size, Zeta potential, entrapment efficiency, stability, and in vitro drug release of GLA-ES were investigated. Tyrosinase activity on melanoma B16-OVA cells were detected to evaluate the inhibition of GLA-ES on the synthesis of melanin, the experiment of potassium ferricyanide reducing power was performed to evaluate the antioxidant effect of GLA-ES, and human epidermal HaCaT cells and rat skin were used for preliminary safety evaluation. Results GLA-ES were yellow translucent liquid, containing vesicular phospholipid bilayer structure, the average particle size of GLA-Et-ES, GLA-PG-ES and GLA-MA-ES were (34.24±0.29), (62.31±1.66) and (41.20±1.13) nm, respectively; The Zeta potential were (-41.0±1.8), (-32.9±0.2) and (-35.8±1.6) mV, the entrapment efficiency were (91.47±2.39)%, (87.33±1.31)% and (91.39±3.59)%, respectively, which had good stability of storage at 4℃ for 20 d, in vitro drug release behaviors of GLA-ES fitted Higuchi equation, implying their sustained release properties. Compared with the glabridin suspension, the inhibitory effects of GLA-Et-ES, GLA-PG-ES and GLA-MA-ES on tyrosinase activity in melanoma B16-OVA cells were increased by 38.07%, 19.58% and 40.42%, respectively. The results of potassium ferricyanide reducing power also showed that GLA-ES had a stronger in vitro antioxidant effect than the glabridin suspension; GLA-ES were nearly nontoxic on normal cells and had no irritation to rat skin. Conclusion GLA-ES can be obtained by hree kinds of softeners, which can inhibit the synthesis of melanin and enhance the antioxidant effect with good safety. The present research will provide the basis for further developing skin-whitening cosmetics or pharmaceutical external preparation. For the insoluble drugs such as glabridin, when mixed alcohol (ethanol-propylene glycol) was selected as the softener to prepare ethosome, it exhibited better encapsulation efficiency and stability than that of ethanol or propylene glycol as the softener alone.

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上海市卫计委中医药科技创新项目（ZYKC201701002）；上海市科委优秀学术带头人资助项目（19XD1423700）