目的 制备具有防治增生性瘢痕（HS）作用的载丹酚酸B的穿膜肽TAT修饰脂质体（SAB-TAT-LIP），建立其质量评价方法，并初步考察其对体外人皮肤成纤维（HSF）细胞增殖和迁移的影响。方法 采用pH梯度逆向蒸发法制备脂质体，超滤法测其包封率，以包封率为评价指标，采用Box-Behnken响应面法优化脂质体的处方工艺；考察其形态、粒径、Zeta电位、体外释放、体外透皮吸收和稳定性等理化性质；在此基础上采用MTT法考察其对HSF细胞增殖的作用，采用划痕法和Transwell小室法考察其对HSF细胞迁移和侵袭的作用。结果 SAB-TAT-LIP的药物包封率为（86.70±0.85）%，平均粒径为（219.90±5.09） nm，Zeta电位（-9.25±0.92） mV，体外24 h累积释放率为62.49%，无突释效应，体外32 h皮肤累积透过率为17.21%，透过速率为（28.33±4.9）μg/（cm²·h），真皮层滞留量为（44.39±6.87）μg/cm²，4℃放置10 d稳定性良好。SAB-TAT-LIP能够显著地抑制HSF细胞的增殖、迁移和侵袭，与对照组相比，差异显著（P<0.01）。结论 优化得到的SAB-TAT-LIP包封率较高、粒径较小，体外释放和透皮行为均满足局部透皮给药制剂的体外释放和透皮规律，对体外HSF细胞的增殖、迁移和侵袭具有抑制作用。

Objective To prepare the liposomes of salvianolic acid B modified with cell penetrating peptide TAT (SAB-TAT-LIP), of which has effects on preventing and treating hypertrophic scars (HS), and establish the method of quality evaluation, as well as preliminarily investigate the effect on the proliferation and migration of human skin fibroblasts (HSF).Methods Liposomes were prepared by pH gradient reverse-phase evaporation method, and the entrapment efficiency was measured by ultrafiltration. Box-Behnken design was performed to optimize the formulation of liposomes by using encapsulation rate as evaluating index. The physicochemical properties of liposomes including morphology, entrapment efficiency, particle size, zeta potential, in vitro release and transdermal absorption, and stability were studied. In addition, the effect of liposomes on proliferation of HSF was examined by MTT assay, and the effect of liposomes on migration of HSF was investigated by scratching method and Transwell assay.Results Based on the optimal formulation of SAB-TAT-LIP, the entrapment efficiency of salvianolic acid B was (86.70 ±0.85)%, the average particle size was (219.90 ±5.09) nm, and the zeta potential was (-9.25 ±0.92). The in vitro 24 h cumulative release was 62.49% of the total drug with no burst effect. The in vitro 32 h cumulated skin penetration rate was 17.21%, the permeance rate was (28.33 ±4.9) μg/(cm²·h), and the retention volume of dermis was (44.39 ±6.87) μg/cm². The stability was good when placed at 4℃ for 10 d. The in vitro cell studies showed that SAB-TAT-LIP can significantly inhibit the proliferation, migration and invasion of human skin fibroblasts, compared with the control group (P < 0.01).Conclusion The optimized SAB-TAT-LIP have higher encapsulation efficiency, smaller particle size, good sustained release effect, and good dermal retention effect which all satisfy the in vitro release and transdermal regulation of local transdermal preparation, and it can significantly inhibit the proliferation, migration and invasion of human skin fibroblasts in vitro.

广东省高等学校优秀青年教师培养计划项目（YQ2015099）；广东省自然科学基金资助项目（2018A0303130234）