目的:明确遗忘型轻度认知功能障碍(aMCI)患者太极拳训练释放的循环外泌体对由Aβ1-42O损伤的人源iPS细胞诱导的神经祖细胞(NPCs)的细胞活性保护作用.方法:招募aMCI受试者随机分为太极拳组和健康教育组,每组各50例,太极拳组进行为期3个月的太极拳训练干预.干预结束后,提取100例受试者外周血的循环外泌体并进行鉴定.通过人源iPS细胞诱导分化得到NPCs并进行免疫荧光鉴定,采用Aβ1.42O建立NPCs损伤模型.在NPCs损伤模型中加入循环外泌体(15、30μg/ml)干预48h,并通过免疫荧光观察NPCs的摄取情况.通过CCK-8观察循环外泌体对Aβ1-42O损伤NPCs的细胞存活率的影响.结果:①TEM、NTA、Western Blot 3个实验结果证明提取的颗粒物为循环外泌体;②免疫荧光结果显示,人源iPS细胞对多能性标志蛋白SOX2、OCT4和NANOG均呈阳性表达,经诱导分化的NPCs对标志蛋白Nestin呈阳性表达,循环外泌体能被NPCs摄取;③CCK-8结果显示,与Aβ,.42O+PBS组相比,太极拳组循环外泌体组NPCs的细胞存活率增加(P＜0.01).结论:太极拳训练释放的循环外泌体可能能够改善Aβ1-42O损伤的人源iPS细胞诱导的神经祖细胞的细胞活性.
Protective effect of circulating exosomes released by Tai Chi training in amnestic mild cognitive impair-ment patients on Aβ1-42O-induced damage in Human iPS cell-derived neural progenitor cells
Objective:To investigate the protective effect of circulating exosomes released by Tai Chi training in amnestic mild cognitive impairment(aMCI)patients on the cell viability of neural progenitor cells(NPCs)induced from human iPS cells and damaged by Aβ1-42 oligomers(Aβ1-42O).Method:A total of 100 aMCI participants were recruited and randomly assigned into two groups:a Tai Chi group and a health education group,with 50 participants in each group.The Tai Chi group underwent a 3-month Tai Chi training intervention.After the intervention,circulating exosomes were extracted and identified from the peripheral blood of the 100 participants.NPCs were induced and differentiated from human iPS cells and identified by immunofluorescence.An NPC damage model was established using Aβ1-42O.Subsequently,cir-culating exosomes(15,30μg/ml)were added to the damaged NPC model for 48 hours,and the uptake of exosomes by NPCs was observed via immunofluorescence.The effect of circulating exosomes on the cell sur-vival rate of Aβ1-42O-damaged NPCs was evaluated by the CCK-8 assay.Result:Transmission electron microscopy(TEM),nanoparticle tracking analysis(NTA),and Western Blot re-sults confirmed that the extracted particles were circulating exosomes;Immunofluorescence results showed that human iPS cells positively expressed pluripotency markers SOX2,OCT4,and NANOG,while the induced NPCs positively expressed the marker Nestin.Circulating exosomes were taken up by NPCs;CCK-8 results indi-cated that compared to the Aβ1-42O+PBS group,the cell survival rate of NPCs in the Tai Chi group treated with circulating exosomes increased significantly(P＜0.01).Conclusion:Circulating exosomes released by Tai Chi training may improve the cell viability of neural progen-itor cells induced from human iPS cells and damaged by Aβ1-42O,suggesting a potential therapeutic effect in neuroprotection.
万方数据
