目的 制备丹参酮II_A（tanshinone II_A，TSA）和丹酚酸B（salvianolic acid B，SAB）共载脂质体（TSA-SAB脂质体），并考察其对人皮肤成纤维细胞（human skin fibroblasts，HSF）增殖的抑制作用。方法 薄膜分散-pH梯度法制备出TSA-SAB脂质体，对其形态、粒径分布、ζ电位、包封率、初步稳定性、体外释药行为和透皮性能等进行表征；CCK-8法、细胞划痕法和Transwell小室法分别考察TSA-SAB脂质体对HSF细胞增殖、迁移和侵袭能力的影响；Western blotting法考察其对基质金属蛋白酶2（matrix metalloproteinase 2，MMP2）和I型胶原（type I collagen，COL-I）蛋白表达水平的影响。结果 TSA-SAB脂质体粒径为（189.50±1.57）nm，ζ电位为（−18.73±1.41）mV，粒子分散系数为0.246±0.030，TSA和SAB包封率分别为（87.93±0.97）%和（91.20±0.47）%，稳定性良好，表现出持续缓慢的释药行为和良好的透皮性能，且能抑制HSF细胞的增殖、迁移和侵袭（P＜0.01），同时显著增加MMP2的合成，降低COL-I的表达（P＜0.01）。结论 所制备的TSA-SAB脂质体粒径分布均匀，稳定性良好，包封率高，可延缓释放速率，提高药物的透皮性，对HSF细胞的增殖、迁移和侵袭均有较好的抑制作用，同时能提高MMP2的表达，抑制COL-I的过度生成，表明TSA-SAB脂质体可能对增生性瘢痕有防治作用。

Objective Tanshinone II_A (TSA) and salvianolic acid B (SAB) co-loaded liposomes (TSA-SAB liposomes) were prepared and their inhibitory effects on the proliferation of human skin fibroblasts (HSF) were investigated. Methods TSA-SAB liposomes were prepared by the thin film dispersion method combined with the pH gradient active drug loading method, and characterized by morphology, particle size distribution, ζ potential, encapsulation rate, preliminary stability, release regularity and transdermal in vitro; The effects of TSA-SAB liposomes on the proliferation, migration and invasion of HSF cells were investigated by CCK-8, cell scratching and Transwell, respectively. And the Effect of TSA-SAB liposomes on the protein expression of matrix metalloproteinase 2 (MMP2) and type I collagen (COL-I) was investigated by Western blotting. Results The TSA-SAB liposomes had a particle size of (189.50 ±1.57) nm, a ζ potential of (−18.73 ±1.41) mV, a particle dispersion coefficient (PDI) of 0.246 ±0.030, and TSA and SAB encapsulation rates of (87.93 ±0.97)% and (91.20 ±0.47)%, respectively. The stability of liposomes was high, and it had sustained drug release and good transdermal property in vitro, which were able to inhibit the proliferation, migration and invasion of HSF cells (P < 0.01), and significantly increased the protein synthesis of MMP2 and decreased the protein expression of COL-I (P < 0.01). Conclusion The TSA-SAB liposomes with uniform particle size distribution, good stability, high encapsulation rate, good transdermal absorption, which inhibited the proliferation, migration and invasion of HSF cells, and also increased the activity of MMP2 collagen degrading enzyme and inhibited the excessive production of COL-I, indicating that TSA-SAB liposomes may have a preventive effect on the generation of hypertrophic scar.

R283.6

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