橄榄苦苷与Aβ1-42的亲和效应及对Aβ1-42诱导的Neuro-2a细胞凋亡的保护作用

摘要

目的 探讨橄榄苦苷对β淀粉样蛋白1-42(amyloid β1-42,Aβ1-42)的抑制能力和亲和效应,以及其对Aβ1-42诱导的Neuro-2a细胞凋亡的保护作用.方法 通过37 ℃孵育制备Aβ1-42纤维,利用Th-T荧光检测橄榄苦苷抑制Aβ1-42纤维的能力,用生物膜层干涉(biolayer interferometry,BLI) 技术实时检测橄榄苦苷与Aβ1-42纤维的亲和效应.进一步用MTT和流式细胞技术检测橄榄苦苷对Aβ1-42诱导的Neuro-2a细胞毒性和凋亡的影响.结果 橄榄苦苷降低Aβ1-42与Th-T反应的荧光强度,抑制Aβ1-42纤维.另外BLI检测显示其与Aβ1-42具有较好的亲和力.并且,橄榄苦苷成剂量依耐性地减轻Aβ1-42诱导的Neuro-2a细胞毒性,降低Aβ1-42诱导的细胞凋亡.结论 研究结果表明,橄榄苦苷可能通过结合并抑制Aβ1-42纤维保护阿尔茨海默病的细胞凋亡.%Objective To investigate the inhibitory and affinity effects of Oleuropein with amyloid beta 1-42 (Aβ1-42) and its protective effect on Aβ1-42-induced cell apoptosis on Neuro-2a. Methods Aβ1-42 fibers were prepared by in-cubation at 37 ℃,Th-T fluorescence was used to detect the ability of Oleuropein to inhibit Aβ1-42 fibers. Biolayer interfer-ometry (BLI) was used to detect the affinity of Oleuropein on Aβ1-42 fibers in real time,and the protective effect of Oleu-ropein on Neuro-2a cytotoxicity and apoptosis induced by Aβ1-42 were verified further. Results Oleuropein decreased the fluorescence intensity of Aβ1-42 reacted with Th-T,inhibited Aβ1-42 fiber. In addition,BLI assay showed that Oleuropein had a good affinity with Aβ1-42. Further experimental results on Neuro-2a showed that Oleuropein can attenuated the cyto-toxicity and decreased the apoptosis rate induced by Aβ1-42 in a dose-dependent manner. Conclusion The results suggest that Oleuropein may protect AD from cell apoptosis by binding and inhibiting the Aβ1-42 fibers.