目的 制备槲皮素与microRNA-150（mR150）共载阳离子固体脂质纳米粒（Que/mR150 SLNs），考察其制备工艺，并评价其体外释放、细胞摄取能力以及眼部给药安全性。方法 采用薄膜分散法制备包载槲皮素的阳离子固体脂质纳米粒（Que-SLNs），以平均粒径、多分散指数（PDI）、包封率为指标，优化其制备工艺；采用静电吸附法将mR150共载于纳米粒中，制备Que/mR150 SLNs，通过琼脂糖凝胶电泳实验考察纳米粒对miRNA的吸附效率；并考察Que/mR150 SLNs中槲皮素的体外释药性能；采用MTT法测定Que/mR150 SLNs对人脐静脉血管内皮细胞HUVEC增殖的影响，并对其进行荧光标记，观察其在HUVEC细胞中的摄取情况；并通过兔眼病理组织切片考察Que/mR150 SLNs对兔眼的刺激性。结果 经过工艺优化，制得的阳离子纳米Que-SLNs载药性、粒径分布、稳定性均较好，其外观呈类球形，放置2个月能保持稳定，槲皮素包封率为（85.25±1.29）%，载药量（1.67±0.02）%，平均粒径（110.00±2.10）nm，Zeta电位（53.20±5.12）mV；体外药物释放结果表明，槲皮素在纳米粒中释放较缓慢，48 h内累积释放量约（80.69±1.29）%；在不同阳离子材料双十八烷基二甲基溴化铵与mR150的质量比（DDAB/RNA）为6：1时，阳离子固体脂质纳米粒可基本将mR150包载完全，且对其粒径、电位影响较小；MTT实验表明，50～150 mg/L的空白纳米质量浓度对HUVEC细胞无明显增殖毒性；细胞摄取实验表明，Cy5与香豆素6（coumarin-6，C6）双荧光标记共载纳米能有效进入HUVEC细胞；兔眼病理组织切片显示Que/mR150 SLNs多次给药对眼部角膜组织无明显损伤。结论 Que/mR150 SLNs固体脂质纳米粒制备工艺稳定可靠、重复性好、贮藏稳定性、生物安全性好，有利于高效递送槲皮素与mR150进入HUVEC细胞，为年龄相关性黄斑变性等血管增生相关疾病的治疗提供思路。

Objective To prepare the cationic solid lipid nanoparticles (Que/mR150 SLNs) co-loaded with quercetin (Que) and microRNA-150 (mR150) and investigate the preparation process, then assess its in vitro release, cell uptake capacity and safety of ocular administration. Method First, thin-film dispersion method was used to prepare quercetin-encapsulated cationic solid lipid nanoparticles (Que-SLNs), and the preparation process was optimized based on the particle size, PDI and encapsulation rate; Using electrostatic adsorption method to co-load mR150 in nanoparticles (Que/mR150 SLNs), and the adsorption efficiency of the miRNA by the nanoparticles was examined by agarose gel electrophoresis experiment; The in vitro release performance of quercetin in Que/mR150 SLNs was investigated; The effect of Que/mR150 SLNs on the proliferation of HUVEC of human umbilical vein endothelial cells was measured by MTT method, and fluorescence labeling was used to observe their uptake in HUVEC; And the irritancy of Que/mR150 SLNs to rabbit eyes was examined by pathological tissue sections of rabbit eyes. Result After process optimization, the cationic nano Que-SLNs had good drug-loading, particle size distribution and stability. The appearance of the cationic nano-Que-SLNs was spherical, and it could be kept stable for two months. The quercetin encapsulation rate was (85.25±1.29)%, the drug load was (1.67±0.02)%, the average particle size was (110.00±2.10) nm, and the Zeta potential is (53.2±5.12) mV; The in vitro drug release results showed that the release of quercetin in the nanoparticles was slow, and the cumulative release amount within 48 h was about (80.69±1.29)%; When the mass ratio of dioctadecyl dimethyl ammonium bromide to mR150 (DDAB/RNA) of different cationic materials was 6:1, the cationic solid lipid nanoparticles basically encapsulated mR150 completely with little effect on its particle size and potential. MTT experiments showed that blank nanometer mass concentration of 50-150 mg/L had no significant proliferation toxicity on HUVEC cells; Cell uptake experiments showed that Cy5 and coumarin-6 dual fluorescently labeled and co-loaded nanometers could effectively enter HUVEC cells; Pathological tissues of rabbit eyes showed that Que/mR150 SLNs had no obvious damage to the eyes. Conclusion The preparation process of Que/mR150 SLNs solid lipid nanoparticles is stable and reliable, with good reproducibility, storage stability and good biological safety, which is conducive to the efficient delivery of quercetin and mR150 into HUVEC cells, which provides the ideas for the treatment of diseases related to angiogenesis

R283.6

四川省科技厅应用基础研究（2019YJ0661）；四川省科技创新苗子工程（2019061）；四川省医学科研课题计划（S18056）；成都市卫生和计划生育委员会临床药学重点学科