三磷酸腺苷对体外培养大鼠雪旺细胞的作用

The effects of ATP on schwann cell in vitro

目的观察三磷酸腺苷(ATP)对体外培养的大鼠雪旺细胞的影响,探讨ATP促进周围神经再生的作用机制。方法用SD乳鼠坐骨神经双酶消化后的剩余组织块培养雪旺细胞,G- 418纯化后培养24 h,分成5组继续培养10d:ATP组(按不同浓度分4组)和正常组。相差显微镜下观察计数,绘制各自的增殖曲线。在24和72 h对各组细胞进行流式细胞分析。结果0.1 mmol ATP组雪旺细胞的增殖不明显;1、10 mmol组24 h和72 h后处于S期的雪旺细胞明显增多;100 mmol组雪旺细胞数则明显减少;1、10、100 mmol组和对照组的倍增时间分别为6.4、5.6、10.2、8.0 d,差异有统计学意义(P<0.05)。结论ATP能显著地促进雪旺细胞的增殖从而促进外周神经再生,但其作用具有浓度依赖性,高浓度ATP反而抑制雪旺细胞的增殖。

To explore the role and mechanism of ATP in promoting peripheral nerve regeneration. Methods The sciatic nerves of 10 four to five-day-old SD rats were removed. After separating SCs with typsin and collagenase, purified cell with G-418. The purified SCs were divided into five groups：ATP 0.1,1,10,100 mmol and the control group. Following 10 days in vitro,the cells from five group were counted with phase contrast microscope every two days. Flow cytometries were carried out in each group after 24 h and 72 h respectively. Results The doubling time of the control group and 0. lmmol group was about 8.0 days, the 1.0,10 and 100mmol ATP groups were 6.4.5.6 and 10. 2 days. The flow cytomery of the five groups showed that SCs in S phase of the 1.0 mmol and 10 mmol groups were significantly higher than the control group,but the 100mmol group was lower. Conclusion ATP could promote SCs proliferation and thereby facilitate peripheral nerve regeneration, the effect would change with the concentration of ATP.