Gabryel Bozena, Bernacki Jacek
Department of Pharmacology, Medical University of Silesia, Katowice, Poland.
Cell Biol Int. 2009 Jul;33(7):739-48. doi: 10.1016/j.cellbi.2009.04.006. Epub 2009 Apr 23.
We investigated whether the immunosuppressive drugs, FK506 and cyclosporine A, increase BDNF protein and/or mRNA expression in ischemic astrocytes and if an increase could be related to changes in the nuclear expression of p-CREB, p-Erk1/2 and p-Akt. The influence of these immunosuppressants on protein and mRNA levels of TrkB and p75(NTR) receptors was also examined. On day 21, cultures of rat astrocytes were subjected to ischemic conditions simulated in vitro (combined oxygen glucose deprivation, OGD) for 8h and exposed to FK506 (10-1000nM) and cyclosporine A (0.25-10microM). FK506 and cyclosporine A (at 1000nM and 0.25microM, respectively) stimulated the expression and release of BDNF in cultured rat cerebral cortical astrocytes exposed to OGD. The immunosuppressants at these doses simultaneously increased p-CREB and p-Erk1/2 expression in the nuclear fraction of astrocytes. The results RT-PCR and Western blot analysis provided further evidence of a modulating influence of the drugs on the expression of trkB and p75(NTR) genes and their protein products in ischemic astrocytes.
我们研究了免疫抑制药物FK506和环孢素A是否会增加缺血性星形胶质细胞中脑源性神经营养因子（BDNF）的蛋白质和/或mRNA表达，以及这种增加是否与磷酸化环磷腺苷反应元件结合蛋白（p-CREB）、磷酸化细胞外信号调节激酶1/2（p-Erk1/2）和磷酸化蛋白激酶B（p-Akt）的核表达变化有关。我们还检测了这些免疫抑制剂对酪氨酸激酶受体B（TrkB）和p75神经营养因子受体（p75(NTR)）的蛋白质和mRNA水平的影响。在第21天，将大鼠星形胶质细胞培养物置于体外模拟的缺血条件下（联合氧糖剥夺，OGD）8小时，然后分别用FK506（10 - 1000 nM）和环孢素A（0.25 - 10 μM）处理。FK506和环孢素A（分别为1000 nM和0.25 μM）刺激了暴露于OGD的培养大鼠大脑皮质星形胶质细胞中BDNF的表达和释放。这些剂量的免疫抑制剂同时增加了星形胶质细胞核部分中p-CREB和p-Erk1/2的表达。逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹分析结果进一步证明了这些药物对缺血性星形胶质细胞中trkB和p75(NTR)基因及其蛋白质产物表达的调节作用。
