不同活化状态的巨噬细胞对雪旺细胞增殖、分泌神经生长因子功能的影响

Effect of macrophages of different activated states on proliferation and NGF level of Schwann cells

目的观察不同活化状态的巨噬细胞对雪旺细胞（SCs）生理功能的影响。方法分离、培养成年SD大鼠巨噬细胞，利用髓鞘组织（20μg／weⅡ）使其激活；从新生1dSD大鼠坐骨神经内分离SCs（1×10^5个／ml）并培养于Transwell下室。共培养组：激活状态的巨噬细胞加入到Transwell上室；对照组1：Transwell上室内加入未激活的巨噬细胞；对照组2：Transwell上室内加入不含细胞的培养基。通过细胞计数、^3H—TdR掺入法检测SCs增殖情况，Western blot检测SCs表达神经生长因子（NGF）水平。结果共培养组从新生大鼠内获取4．16×10^6个SCs，明显高于对照组1（3．83×10^6）和对照组2（3．19×10^6）（P〈0.05）；3H—TdR结果显示细胞增殖水平（0.2766±0．0079）明显强于对照组1（0．2268±0．0084）和对照组2（0．1929±0．0031）（P〈0．05）；WB结果显示NGF表达水平明显高于对照组（P〈0．05）。结论巨噬细胞对SCs的影响受其活化状态的影响，激活状态的巨噬细胞可以明显促进SCs增殖，增强其表达NGF的能力。

Objective To explore the influence of macrophages of different activated states on physiological function of Schwann cells （SCs）. Methods SCs were obtained from sciatic nerve of neonatal SD rats at day 1, and macrophages were harvested from adult SD rats and activated with myelin （20 μg/well） isolated from peripheral nerve. In co-cultured group, SCs （1 × 10^6/ml） were cultured in the low- er room and activated macrophages were cultured in the upper room of Transwell separately. In control group 1, SCs were co-cultured with un-activated macrophages, and in control group 2, SCs were co-cultured with cell-free culture medium. The proliferative level of SCs was measured by cell count and 3 H-TdR incorporation, and the level of NGF in SCs was detected by Western blot. Results The number and proliferation level of SCs in co-cultured group was 4.16 × 10^6 and 0. 2766 ±0.0079 respectively, which was higher than in control groups 1 （ 3.83 × 10^6, 0. 2268 ± 0. 0084 ） and control group 2 （ 3.19 × 10^6, 0. 1929 ± 0. 0031 ） （ P 〈 0.05 ）. Western blot revealed that the expression level of NGF in co-cultured group was significantly higher than in control groups （ P 〈 0.05 ）. Conclusion The ability of proliferation and level NGF of SCs could be promoted by macrophages activated by myelin.