载miR-144抑制剂的脂质体杂合红细胞膜纳米药物的构建及初步评价OA

Objective To construct a liposomehybrid red blood cell membrane(LHRM)nanode-livery system loaded with microRNA-144(miR-144)inhibitor,and to evaluate its physicochemical properties,biocompatibility,in vivo circulation characteristics,and in vitro delivery capacity.Meth-ods Red blood cell membranes were extracted by hypotonic lysis,liposomes were prepared by thinfilm hydration,and LHRM nanoparticles were fabricated by fusing the two components at various mass ratios.The nanoparticles were characterized by dynamic light scattering(DLS),Zeta potential,and trans-mission electron microscopy(TEM).Membrane fusion was verified by DiO/DiI fluorescence colo-calization assays.The in vivo circulation profile was evaluated by intravenously injecting DiI labeled nanoparticles via the tail vein into C57BL/6 mice.Using mouse erythroleukemia cells(MEL)as an in vitro model,the cellular delivery efficiency and cytotoxicity of nanoparticles with different mass ratios were examined.Furthermore,western blot was performed to detect the expression changes of NF-E2 related factor 2(Nrf2)protein following miR-144 inhibitor delivery.Results The LHRM nanocarriers were successfully constructed,exhibiting a regular core-shell structure with significantly increased particle size compared to red blood cell membranes alone;fluorescence colocalization con-firmed successful membrane fusion.Among the various formulations,the mass ratio of red blood cell membrane to liposome at 6∶1 yielded the optimal cellular delivery efficiency.The preliminary en-capsulation efficiencies of miR-144 inhibitor loaded LHRM and liposomes were(75.05±5.49)％and(60.23±6.85)％,respectively.In in vivo experiments,the LHRM group significantly delayed the blood fluorescence decay trend compared with the liposome group.MTT assay preliminarily indi-cated that LHRM exhibited favorable biocompatibility and low cytotoxicity toward MEL cells.West-ern blot analysis demonstrated that treatment with miR-144 inhibitor loaded LHRM upregulated Nrf2 protein levels in MEL cells.Conclusion The constructed LHRM nanodelivery system possesses fa-vorable structural stability,biocompatibility,and delivery capacity,and demonstrates a superior blood fluorescence decay profile compared to liposomes in preliminary in vivo experiments.The suc-cessful construction of this delivery system provides a biomaterial platform for effective delivery of miR-144 inhibitors and amelioration of oxidative stress in β-thalassemia cells.