目的 合成透明质酸（HA）接枝单油酸甘油酯（GMO）两亲性聚合物HGO，并研究其所制备载阿霉素（DOX）纳米粒的理化性质及体外抗肿瘤效果。方法 HA与GMO通过酯化反应制得载体聚合物HGO，通过核磁共振波谱法及红外光谱法对其进行结构表征；采用芘荧光探针法测定聚合物临界聚集浓度（CAC）。采用透析法制备聚合物HGO载阿霉素（DOX@HGO）纳米粒，并对其进行粒径分布、Zeta电位及微观形态的表征；通过检测其在不同离子强度、不同pH条件下的粒径变化考察纳米粒的体外稳定性；考察DOX@HGO纳米粒在不同pH条件下的体外释放行为；CCK-8法考察DOX@HGO纳米粒对MDA-MB-231细胞的体外抑瘤效果；并通过荧光显微镜研究MDA-MB-231细胞对DOX溶液、DOX@HGO纳米粒的摄取能力，以及HA预处理对DOX@HGO纳米粒摄取的影响。结果 成功制得两亲性聚合物HGO，聚合物HGO中GMO的取代度为15.8%，CAC为0.023 mg·mL^-1。DOX@HGO纳米粒呈规则的球形，平均粒径为（130.800±1.709）nm，平均电位为（-32.600±0.153）mV，包封率和载药量分别为（98.65±0.74）%和（33.03±0.17）%，在不同离子强度下、模拟胃肠液中表现出良好的稳定性；DOX@HGO纳米粒的体外释放表现出pH依赖性。体外抗肿瘤活性实验表明，DOX@HGO纳米粒对MDA-MB-231细胞的生长具有较好的抑制作用；与DOX溶液比较，DOX@HGO纳米粒显著增加肿瘤细胞对于DOX的摄取（P＜0.05） ，HA预处理显著减少肿瘤细胞对DOX@HGO的摄取（P＜0.05）。结论 所构建的DOX@HGO纳米粒具有良好的理化性质，并且具有一定的pH敏感性及靶向抗肿瘤细胞的能力，是具有应用潜力的药物载体。

Objective To prepare an amphiphilic polymer HGO of hyaluronic acid (HA) grafted glyceryl monooleate (GMO), study the physicochemical properties of doxorubicin (DOX) loaded nanoparticles and determine the physicochemical properties as well as in vitro antitumor effect. Methods HA and GMO were esterified to prepare the carrier polymer HGO, and the structure of the polymer was characterized by ¹H-NMR and FT-IR. The critical aggregate concentration (CAC) was determined by pyrene fluorescence method. The hyaluronic acid grafted glyceryl monooleate loaded DOX nanoparticles (DOX@HGO) were prepared by dialysis method, and their particle size distribution, Zeta potential and micromorphology were characterized. The in vitro stability of the nanoparticles was investigated by the particle size changes under different ionic strengths and pH conditions. The drug release behaviors of the nanoparticles were examined under different pH. The in vitro antitumor effect of drug loaded nanoparticles were evaluated by CCK-8 method. The cellular uptake of DOX and DOX@HGO and effect of HA pre-treatment on DOX@HGO nanoparticle uptake was investigated under fluorescence microscope. Results The amphiphilic polymer HGO was successfully prepared, and the substitution degree of GMO in polymer HGO was 15.8%, the CAC was 0.023 mg·mL^-1. DOX@HGO nanoparticles showed a regular spherical shape, with an average particle size of (130.800 ±1.709) nm, an average potential of (-32.600 ±0.153) mV, an encapsulation efficiency of (98.65 ±0.74)% and a drug loading capacity of (33.03 ±0.17)%, respectively. The polymeric nanoparticles showed good stability under different ionic strengths and simulated gastrointestinal fluid dilution. Moreover, the in vitro release of DOX@HGO nanoparticles indicated pH dependent behavior. In vitro antitumor activity experiments showed that DOX@HGO nanoparticles had a good inhibitory effect on the growth of MDA-MB-231 cells and could significantly improve the cellular uptake of DOX by tumor cells. Compared with DOX solution, DOX@HGO nanoparticle significantly increased the uptake of DOX by tumor cells (P < 0.05), and HA pretreatment significantly reduced the uptake of DOX@HGO by tumor cells (P < 0.05). Conclusion The DOX@HGO nanoparticles prepared in this study have good physicochemical properties, excellent pH sensitivity and great anti-tumor targeting effects, which exhibited great application potential in drug delivery systems.

国家自然科学基金面上项目（82074030）；国家自然科学基金青年项目（82104568）；天津市教委科研计划项目（2020KJ195）