α倒捻子素与载α倒捻子素纳米制剂促进小胶质细胞摄取Aβ的作用比较

Comparison in effects of αM and NP-αM on microglial uptake of Aβ

目的探讨研究α倒捻子素(αM)和载αM聚乙二醇聚乳酸纳米制剂(NP-αM)对小胶质细胞摄取β-淀粉样蛋白(Aβ)的作用效果及其机制。方法制备空载纳米制剂(NP)和NP-αM,测量其电位、粒径,并进行透射电镜下纳米制剂表征分析;分别使用NP、αM、NP-αM(包含与前两组相同浓度的NP或αM)处理BV2细胞和原代小胶质细胞,采用高内涵扫描系统定量检测不同处理组细胞摄取的羧基荧光素标记Aβ;使用香豆素6(Cou6)分别标记NP和NP-αM,并以其分别作用BV2细胞和原代小胶质细胞。采用共聚焦显微镜和高内涵扫描系统检测各组细胞摄取纳米粒的量。结果①纳米制剂稳定、大小均匀、分散均一;②NP对细胞摄取Aβ无效果,αM和NP-αM均显著促进了小胶质细胞摄取Aβ,NP-αM的作用更显著(P均<0.05);③与NP比较,αM、NP-αM均可以促进小胶质细胞摄取纳米制剂(P均<0.05)。结论NP-αM较αM可以更好地促进小胶质细胞摄取Aβ,此有可能为阿尔茨海默病的致病蛋白Aβ清除提供一种纳米疗法。

Objective To compare the effects ofα-mangostin(αM)andαM-loaded nanoparticle(NP-αM)on microglial uptake of amyloidβ-protein(Aβ)and to reveal the mechanism.Methods The drug-free nanoparticle(NP)and NP-αM were prepared,and their zeta potential and particle size were measured.The nanoparticles were also characterized by transmission electron microscopy.BV2 cells and primary microglia were treated with NP,αM,and NP-αM(containing the same concentrations of NP orαM as the former two groups),respectively,then the cells were incubated with carboxyfluorescein-labeled Aβ,and the cellular uptake of Aβwas measured with a high-content scanning system.NP and NP-αM were labeled with Cou6(Cou6-NP and Cou6-NP-αM,respectively).BV2 cells and primary microglia were treated with Cou6-NP and Cou6-NP-αM,respectively,and the cellular uptake of the nanoparticles was measured via confocal microscopy and high-content scanning analysis.Results①The prepared nanoparticles were stable,uniform in size and monodispersed.②NP did not affect the microglial uptake of Aβwhile bothαM and NP-αM significantly promoted the uptake,and NP-αM was more efficient(P<0.05).③compared with NP,bothαM and NP-αM could promote microglial uptake of the nanoparticles(both P<0.05).Conclusion NP-αM can more efficiently promote the uptake of Aβin microglia thanαM,and this may provide a nanomedicine for the clearance of the key pathogenic protein Aβin Alzheimer's disease.