嗜中性粒细胞损害嗅鞘细胞神经修复功能的体外研究

Neutrophil-induced impairment in axonal regeneration of olfactory ensheathing cells in vitro

目的体外研究嗜中性粒细胞对嗅鞘细胞(OECs)功能的影响。方法从流产人胚胎中分别培养OECs、星形胶质细胞,健康志愿者外周血中分离嗜中性粒细胞。6孔板中,1×106个OECs与1×103个嗜中性粒细胞共孵育48h后,观察OECs的细胞形态和凋亡状况,并收集OECs;RT-PCR检测NGF、BDNF及GNDF基因表达情况;通过OECs和星形胶质细胞的相遇培养实验,检测OECs的迁移能力。结果共培养处理后,嗜中性粒细胞促使OECs凋亡,使OECs的NGF、BDNF及GNDF的表达显著下降(P<0.01);在OECs和星形胶质细胞的相遇培养实验中,形成迁移的明显边界。结论嗜中性粒细胞削弱了OECs在神经再生中的能力。

Objective To evaluate the infuences of neutrophils on axonal regeneration of OECs in vitro.Methods OECs and astrocytes were prepared from embryo sample,while neutrophils were purified from peripheral blood of health individuals.Apoptosis of OECs and expression of NGF,BDNF and GDNF were assessed on 2nd day of OECs and neutrophils coculture.The interactions between astrocytes and either neutrophil treated-or untreated-OECs were performed by confrontation assay.Results Neutrophil-induced apotosis of OECs was observed after cocultures.NGF,BDNF and GDNF expression was significantly reduced in experimental group compared with that in control group （P0.01）.Boundary formation in OECs/astrocytes confrontation assays was induced by neutrophils treatment.Conclusion Neutrophils impair the axonal regeneration ability of OECs.