Centre for Neuromuscular and Neurological Disorders, University of Western Australia and Australian Neuromuscular Research Institute, Western Australia, Australia.
Neurosci Res. 2012 Jul;73(3):191-8. doi: 10.1016/j.neures.2012.04.007. Epub 2012 Apr 27.
In this study we have assessed sodium-calcium exchanger (NCX) protein over-expression on cell viability in primary rat cortical neuronal and HEK293 cell cultures when subjected to oxygen-glucose deprivation (OGD). In cortical neuronal cultures, NCX2 and NCX3 over-expression was achieved using adenoviral vectors, and following OGD increased neuronal survival from ≈20% for control vector treated cultures to ≈80% for both NCX isoforms. In addition, we demonstrated that NCX2 and NCX3 over-expression in cortical neuronal cultures enables neurons to maintain intracellular calcium at significantly lower levels than control vector treated cultures when exposed to high (9mM) extracellular calcium challenge. Further assessment of NCX activity during OGD was performed using HEK293 cell lines generated to over-express NCX1, NCX2 or NCX3 isoforms. While it was shown that NCX isoform expression differed considerably in the different HEK293 cell lines, high levels of NCX over-expression was associated with increased resistance to OGD. Taken together, our findings show that high levels of NCX over-expression increases neuronal and HEK293 cell survival following OGD, improves calcium management in neuronal cultures and provides additional support for NCX as a therapeutic target to reduce ischemic brain injury.
在这项研究中，我们评估了钠钙交换蛋白（NCX）在原代大鼠皮质神经元和 HEK293 细胞培养物中受到氧葡萄糖剥夺（OGD）时对细胞活力的过表达作用。在皮质神经元培养物中，使用腺病毒载体实现了 NCX2 和 NCX3 的过表达，OGD 后，与对照载体处理的培养物相比，两种 NCX 同工型的神经元存活率从约 20%增加到约 80%。此外，我们证明，在皮质神经元培养物中过表达 NCX2 和 NCX3 可使神经元在暴露于高（9mM）细胞外钙挑战时将细胞内钙维持在比对照载体处理的培养物低得多的水平。使用过表达 NCX1、NCX2 或 NCX3 同工型的 HEK293 细胞系进一步评估了 OGD 期间的 NCX 活性。虽然表明不同 HEK293 细胞系中 NCX 同工型表达差异很大，但高水平的 NCX 过表达与对 OGD 的抵抗力增加有关。总之，我们的研究结果表明，高水平的 NCX 过表达可增加 OGD 后神经元和 HEK293 细胞的存活率，改善神经元培养物中的钙管理，并为 NCX 作为减少缺血性脑损伤的治疗靶点提供更多支持。
