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

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35 μmol/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)10 μmol/L氟西汀干预星形胶质细胞不同时间后,48 h组细胞培养液GDNF浓度(68±13)fg/L]高于0 h组(32±11)fg/L]、6 h组(34±12)fg/L]、12 h组(41±17)fg/L]、24 h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48 h后,10 μmol/L浓度组的细胞培养液GDNF浓度(64±17)fg/L]高于0 μmol/L(39±15)fg/L]和1 μmoVL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48 h后,撤离氟西汀24 h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24 h后,10 μmol/L和20 μmol/L浓度组细胞GDNFmRNA表达量分别为(0.008 1±0.001 1)和(0.006 3±0.000 3)]高于0 μmol/L、1 μmol/L及5 μmol/L浓度组分别为(0.003 1±0.000 7)、(0.003 9±0.000 3)和(0.004 1±0.000 2)],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.

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

Objective To explore the effects of fluoxetine on glial cell line-derived neurotrophic factor (GDNF) synthesis and release in cultured rat astrocytes. Methods The MTT 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 astrocytes after with draual of fluoxetine for 24 hours, (P<0.01or 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 neuroprotective properties.