Institut für Pharmazeutische und Medizinische Chemie, Westfälische Wilhelms Universität, Hittorfstrabetae 58-62, D-48149 Münster, Germany.
Neuroscience. 2009 Dec 29;164(4):1695-700. doi: 10.1016/j.neuroscience.2009.09.055. Epub 2009 Sep 25.
The neurotrophic and neuroprotective activity of fibroblast growth factor (FGF2) is well documented. In this study, we attempted to demonstrate that binding of ATP to FGF2 is essential for its neuroprotective effect. Incubation of primary cultures of rat embryonic (E18) cortical neurons with alkaline phosphatase decreased the ATP concentration in the culture medium from about 8 to 0.3 nM measured luminometrically. Reduction of ATP concentration below 1 nM abolished the neuroprotective effect of FGF2. However, when the more stable nucleotide triphosphate gammaS-ATP was used which could not be cleaved by alkaline phosphatase, FGF2 still protected the cultured cortical neurons against damage. In control experiments alkaline phosphatase alone did not influence neuroprotection. In addition, also ATPase and apyrase were used as ATP cleaving enzymes. Added to the culture medium, both enzymes were capable of decreasing ATP below the critical level of approximately 1 nM, and the neuroprotective activity of FGF2 was abolished. Thus, our results demonstrate for the first time that the FGF2/ATP complex but not FGF2 alone mediates neuroprotection.
成纤维细胞生长因子 (FGF2) 的神经营养和神经保护作用已有充分的文献记载。在这项研究中，我们试图证明 FGF2 与 ATP 的结合对于其神经保护作用是必不可少的。用碱性磷酸酶孵育大鼠胚胎 (E18) 皮质神经元的原代培养物，可使培养基中 ATP 的浓度从约 8 nM 降低到通过发光法测量的 0.3 nM。当 ATP 浓度降低到 1 nM 以下时，FGF2 的神经保护作用就会消失。然而，当使用碱性磷酸酶无法切割的更稳定的核苷酸三磷酸 γS-ATP 时，FGF2 仍然可以保护培养的皮质神经元免受损伤。在对照实验中，碱性磷酸酶本身并不影响神经保护作用。此外，还使用了 ATP 酶和 apyrase 作为 ATP 切割酶。将这两种酶添加到培养基中，都可以将 ATP 降低到约 1 nM 的临界水平以下，从而使 FGF2 的神经保护活性丧失。因此，我们的研究结果首次证明，是 FGF2/ATP 复合物而不是单独的 FGF2 介导了神经保护作用。
