mRNA脂质纳米粒载药系统的构建及体外评价

目的 构建脂质纳米粒DLin-LNP，以EGFP-mRNA为模型药，考察DLin-LNP对于mRNA的体外递送能力。方法 采用薄膜水化法制备DLin-LNP, 并进一步制备 DLin@mRNA，对纳米粒进行表征，使用激光扫描共聚焦显微镜观察脂质纳米粒胞内的分布情况，以RM-1细胞为模型考察胞内转染情况。结果 成功制备了脂质纳米粒DLin-LNP，其粒径为（151.1±2.1） nm，空载电位为（23.7±0.5） mV。DLin-LNP在RM-1细胞中转染mRNA效率较高，其毒性远低于市售脂质体Lipo8000_，且 DLin-LNP脂质纳米粒稳定性好。结论 DLin-LNP具有高转染效率和安全性，且稳定性好，可作为mRNA递送载体，为后续脂质纳米粒肿瘤治疗中的应用提供依据。

Construction and in vitro evaluation of an LNP system for mRNA delivery

Objective To construct lipid nanoparticles DLin-LNP for mRNA delivery. Methods DLin-LNP was prepared by thin film hydration method, and DLin-LNP/mRNA was further constructed by using EGFP-mRNA as model drug. The particle size, zeta potential, and appearance morphology were measured. Furthermore, the intracellular distribution and transfection of DLin-LNP/mRNA in RM-1 cells was investigated by laser scanning confocal microscope. Results DLin-LNP was successfully prepared. The average particle size was about (151.1±2.1) nm, the no-load potential was (23.7±0.5) mV. The cytotoxicity of DLin-LNP was far lower than that of the commercially available liposomal Lipo8000. The results of transfection experiment indicated that DLin-LNP has high transfection efficiency for mRNA delivery with low cytotoxicity and good stability. Conclusion DLin-LNP could become a potential mRNA vector for gene therapy.