目的 构建一种具有肿瘤双靶向功能的仿脂蛋白结构纳米载体（FA-BSA-LC/SPC），共载紫杉醇（paclitaxel，PTX）及白藜芦醇（resveratrol，Rev），用于抗多药耐药肿瘤研究。方法 以牛血清白蛋白（bovine serum albumin，BSA）作为构建仿脂蛋白结构纳米载体的蛋白部分，叶酸（folic acid，FA）通过偶联在BSA上形成偶联蛋白FA-BSA，与载药脂质核心部分（LC/SPC-PTX+Rev）通过静电作用相结合；利用透析法考察载药纳米制剂的体外释药行为；通过纳米制剂在血浆中的凝聚变化对其血浆稳定性进行考察；采用MTT法对载药纳米制剂进行多药耐药细胞毒性研究；利用激光扫描共聚焦显微镜考察仿脂蛋白结构纳米载体肿瘤双靶向效果。结果 制备的纳米制剂具有较高的紫杉醇及白藜芦醇包封率（分别为91.02%、91.30%），适用于药物紫杉醇-白藜芦醇（4：1）的共载；载药纳米制剂在体外具有明显的缓释药物行为；在血浆孵育后的粒径分布没有明显变化，表明其在血浆中具有良好的稳定性；通过细胞毒性研究发现FA-BSA-LC/SPC-PTX+Rev对多药耐药肿瘤细胞具有明显的细胞生长抑制作用；通过细胞摄取实验结果表明，FA-BSA-LC/SPC具有明显的肿瘤双靶向功能。结论FA-BSA-LC/SPC-PTX+Rev作为具有生物相容性、肿瘤双靶向功能的一种仿脂蛋白结构纳米载体共载药物紫杉醇及逆转剂白藜芦醇，达到理想的治疗多药耐药肿瘤效果。

Abstract:Objecctive To construct dual-targeting lipoprotein-mimic nanocarrier (FA-BSA-LC/SPC) with paclitaxel (PTX) and resveratrol (Rev) co-loaded for reversing multidrug resistance (MDR) activity. Methods The bovine serum albumin (BSA) was used as the protein model for constructing the lipoprotein-mimic nanocarrier, the further conjugating folic acid to BSA generated modified BSA (FA-BSA) and then coated the surface of the lipid core (LC/SPC-PTX+Rev) through electrostatic attraction; The in vitro drug release study was conducted using dialysis method; The stability of FA-BSA-LC/SPC-PTX+Rev was studied by testing the aggregation in 50% human plasma. The cytotoxicity of drug loaded nanocarriers against MDA-MB-231/taxol cells were evaluated by the standard MTT assay. The in vitro dual-targeting ability was studied by laser scanning confocal microscopy (LSCM). Results FA-BSA-LC/SPC-PTX+Rev had high encapsulation efficiency of PTX and Rev (91.02% and 91.30%, respectively), which was suitable for PTX and Rev delivery; In vitro drug release study showed that drug released from nanocarrier was significantly prolonged. The stability study showed that there was no significant change, suggesting that the coupling protein could increase the stability in plama. The cytotoxicity studies demonstrated that FA-BSA-LC/SPC-PTX+Rev had greater cellular inhibition in MDA-MB-231/taxol cells; The cellular uptake studies showed that FA-BSA-LC/SPC exhibited excellent dual-targeting activity. Conclusion FA-BSA-LC/SPC-PTX+Rev, as biocompatible and dual tumor-targeting nanocarrier, is a promising system with PTX and Rev loaded for optimal reversing MDR efficacy.

R283.6

山东省自然科学基金青年项目（ZR2020QH325）；大学生创新创业训练项目（S202110452120）