氟西汀对大鼠星形胶质细胞分泌的胶质源性神经营养因子的影响

Fluoxetine stimulates glial cell fine-derived neurotrophic factor synthesis and release in cultured rat astrocytes

目的探讨氟西汀对大鼠星形胶质细胞分泌的胶质源性神经营养因子（GDNF）的影响。方法以氟西汀干预体外培养的大鼠海马星形胶质细胞，通过四甲基偶氮唑盐法（MTT）检测不同浓度氟西汀对细胞活力的影响；采用酶联免疫吸附测定法（ELISA）检测细胞培养液GDNF浓度及Real-time PCR法检测GDNFmRNA的表达。结果（1）氟西汀浓度超过35μmol/L浓度时，可降低细胞活性，差异有统计学意义（P〈0．01或P〈0．05）；（2）10μmol／L氟西汀干预星形胶质细胞不同时间后，48h组细胞培养液GDNF浓度[（68±13）fg／L]高于0h组[（32±11）fg/L]、6h组[（34±12）fg/L]、12h组[（41±17）fg/L]、24h组[（45±13）fg/L]，差异均有统计学意义（P均〈0．01）；（3）不同浓度氟西汀作用星形胶质细胞48h后，10μmol/L浓度组的细胞培养液GDNF浓度[（64±17）fg/L]高于0μmol／L[（39±15）fg/L]和1μmol/L浓度组[（39±18）fg/L]，差异均有统计学意义（P均〈0．05）；（4）氟西汀作用星形胶质细胞48h后，撤离氟西汀24h后星形胶质细胞仍明显分泌GDNF，差异有统计学意义（P〈0．01或P〈0．05）；（5）不同浓度氟西汀作用星形胶质细胞24h后，10μmoL／L和20μmol/L浓度组细胞GDNF mRNA表达量[分别为（0．0081±0．0011）和（0．0063±0．0003）]高于0μmol／L、1μmol／L及5μmoL／L浓度组[分别为（0．0031±0．0007）、（0．0039±0．0003）和（0．0041±0．0002）]，差异均有统计学意义（P均〈0．01）。结论氟西汀可能通过促进星形胶质细胞GDNF的分泌来发挥其神经保护作用。

Objective To explore the effects of fluoxetine on glial cell line-derived neurotrophic factor （GDNF） synthesis and release in cultured rat astrocytes. Methods The MTr assay were used to evaluate cell viability, the expression of GDNF mRNA were detected with real-time PCR. The level of GDNF in cell-conditioned media was measured using enzyme linked immunosorbent assay （ELISA）. Results （1） The cell viability was significantly decreased when the concentration of fluoxetine was more than 35 μmol/L （ P 〈 0. 01 or P 〈 0. 05 ）. （2） When the cells were treated with the 10 μmol/L fluoxetine for different time, the level of GDNF in cell-conditioned media of the 48 hour group [ （68 ± 13 ） fg/L] was significantly higher than that of the 0, 6, 12, 24 hour group [ （32 ± 11）, （34 ±12）, （41 ± 17） and （45 ± 13） fg/L] （P 〈 0.01 ）. （3） When the cells were treated with the different concentration of fluoxetine for 48 hour, the level of GDNF in cell-conditioned media of 10 μmol/L group[ （64 ± 17） fg/L] was significantly higher than that of 0 μmol/L and 1 μmol/L groups [ （ 39 ± 15 ）, （ 39 ± 18 ） fg/L ] （ P 〈 0. 05 ）. （4） When the cells were treated at a range of concentrations of fluoxetine for 48 hour, the GDNF was also significantly released by astroeytes after with draual of fluoxetine for 24 hours, （ P 〈 0.01 or P 〈 0.05 ）. （ 5 ） After the cells were treated with the different concentrations of fluoxetine for 24 hour, the expression of GDNF mRNA in 10 μmol/L and 20 μmoL/L groups [ （0. 008 1 ± 0. 001 1 ）, （ 0. 006 3 ±0. 000 3 ） ] were significantly higher than that in 0 μmol/L, 1 μmol/L and 5 μmol/L groups [（0.003 1 ±0.000 7）, （0.003 9 ±0.000 3）, （0. 004 1 ± 0. 000 2） ]. Conclusion The results suggest that fluoxetine could stimulate GDNF release from astrocytes, which might underlie it＇s neuroproteetive properties.