红细胞生成素对体外培养许旺细胞增殖周期及其分泌神经营养因子的影响

Effects of erythropoietin on cell cycle and neurotrophic factor secretion of Schwann cells in vitro

目的观察促红细胞生成素（EPO）对体外培养许旺细胞（SCs）增殖周期及其分泌神经生长因子（NGF）和睫状神经生长因子（CNTF）的影响，探讨EPO促进周围神经再生的机制。方法对成年新西兰兔的坐骨神经进行结扎预变性，1周后取预变性的坐骨神经，剥净神经外膜，用植块法、差速贴壁法培养纯化许旺细胞，向培养基中加入不同剂量的EPO，通过流式细胞术检测细胞增殖周期分布，应用ELISA测定培养上清液中的NGF和CNTF水平。结果与对照组相比，实验组许旺细胞处于S期数量（S％）和增殖指数PrI值[（S＋G2M）％]都有明显升高，培养上清液中的NGF和CNTF水平明显增高。结论EPO可提高体外培养许旺细胞的增殖活性，并且能提高许旺细胞分泌神经营养因子的水平，这可能是其促进周围神经再生的作用机制之一。

Objective To observe the effects of EPO on cell cycle and the secretion of NGF and CNTF of SCs in vitro and explore how EPO improve peripheral nerve regeneration. Methods The sciatic nerves of the adult New Zealand rabbits were ligated and pre - degenerated. After a week, the pre - degenerated nerves were fetched and their ectoblastics were removed. We adopted a method of trasplant piece and speed difference adhestion to cultivte and purify SCs. Different concentrations of EPO were added into cutures of purified SCs respectively. The cell cycle distribution of SCs.was observed by flow cytometry,and the level of NGF and CNTF in the culture was detected by ELSIA. Results In the EPO supplemention group,the number of SCs in S phase （S%） was significantly increased, G1% obviously decreased and the PrI [ （S ＋ G2 M）% ] enhanced when the SCs were stimulated. Conclusions EPO enhance SCs proliferation activity and increase the secretion of neurotrophic factors, which may be explain how EPO improve peripheral nerve regeneration.