血小板膜仿生纳米粒用于肺靶向基因递送的构建及靶向性评价

Platelet membrane biomimetic nanoparticles for lung-targeted gene delivery: construction and targeting evaluation

摘要

目的 构建负载 siHIF-1α 与 PSS31 的血小板膜仿生纳米粒, 实现低氧性肺动脉高压的高效治疗。方法 采用静电吸附法和挤压法制备 siHIF-1α/PSS31 及 siHIF-1α/PSS31@PM; 采用激光粒度测定仪、透射电镜、凝胶电泳等表征其粒径、电位、形态及稳定性等; 通过溶血实验和细胞毒性实验评估其体外生物相容性; 通过流式细胞仪等检测常氧、低氧损伤肺微血管内皮细胞 (PMVECs) 及肺动脉平滑肌细胞 (PASMCs) 对 siHIF-1α/PSS31 及 siHIF-1α/PSS31@PM 的摄取效率; 通过活体成像及免疫荧光染色实验考察其体内靶向性。结果 阳离子聚合物 PSS31 可高效负载 siHIF-1α。siHIF-1α/PSS31@PM 的粒径约为 (180.6 ± 0.5)nm, 形态为球形, 具有明显的核壳结构, 且在 6 d 内粒径无显著变化。siHIF-1α/PSS31@PM 无明显溶血现象, 对 PMVECs 和 PASMCs 的细胞活力没有明显影响。siHIF-1α/PSS31@PM 能够靶向低氧损伤的肺组织, 并蓄积于 PMVECs 及 PASMCs 中, 靶向效果显著优于 siHIF-1α/PSS31 (P<0.05)。结论 siHIF-1α/PSS31@PM 能够高效靶向低氧损伤 PMVECs 和 PASMCs, 为基因药物的肺靶向递送提供了新策略。

Abstract

Objective To construct platelet membrane biomimetic nanoparticles loaded with siHIF-1α and PSS31 for efficient treatment of hypoxic pulmonary hypertension. Methods The lung-targeted biomimetic nanoparticles siHIF-1α/PSS31 and siHIF-1α/PSS31@PM were prepared using electrostatic adsorption and extrusion methods. Their particle size, zeta potential, morphology, and stability were characterized by laser particle size analyzer, transmission electron microscopy, and gel electrophoresis. In vitro biocompatibility was evaluated through hemolysis and cytotoxicity assays. The uptake efficiency of siHIF-1α/PSS31 and siHIF-1α/PSS31@PM by normoxic and hypoxically damaged pulmonary microvascular endothelial cells (PMVECs) and pulmonary artery smooth muscle cells (PASMCs) was assessed using flow cytometry. In vivo targeting ability was investigated via live imaging and immunofluorescence staining experiments. Results The cationic polymer PSS31 efficiently encapsulated siHIF-1α. siHIF-1α/PSS31@PM exhibited a spherical morphology with a distinct core-shell structure and an average particle size of approximately (180.6 ± 0.5) nm, showing no significant change in size over a period of 6 d. Besides, siHIF-1α/PSS31@PM showed no obvious hemolysis phenomenon and had no significant effect on the cell viability of PMVECs and PASMCs. Furthermore, siHIF-1α/PSS31@PM effectively targeted hypoxia-injured lung tissue and accumulated in PMVECs and PASMCs, with significantly better targeting efficiency than siHIF-1α/PSS31 (P<0.05). Conclusion siHIF-1α/PSS31@PM demonstrates highly efficient targeting of hypoxically damaged PMVECs and PASMCs, thereby offering a novel strategy for the pulmonary-targeted delivery of gene-based therapeutics.

关键词

仿生基因递送载体 / 血小板膜 / 阳离子聚合物 / 低氧诱导因子 1-α / 小干扰 RNA / 低氧性肺动脉高压 / 肺靶向递药系统 / 基因肝外递送

Key words

biomimetic gene delivery carrier / platelet membrane / cationic polymer / hypoxia-inducible factor-1alpha / small interfering RNA / hypoxic pulmonary hypertension / lung-targeted drug delivery system / extrahepatic gene delivery

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基金资助

陕西省中医药管理局项目(SZY-KJCYC-2025-ZY-013)