Department of Bio-Medical Sciences, Section of Physiology, University of Catania, Catania, Italy.
J Neurosci Res. 2013 Feb;91(2):313-20. doi: 10.1002/jnr.23154. Epub 2012 Nov 22.
The present study analyzed the in vitro effects induced by sodium L-lactate on human astrocytes and the SH-SY5Y cell line, when added at concentrations of 5, 10, and 25 mmol/liter. Expression of brain-derived neurotrophic factor (BDNF), inducible nitric oxide synthase (iNOS), and heat shock protein 70 kDa (HSP70) was evaluated by Western blot analysis. Cell viability with MTT, release of nitric oxide (NO) through the Griess reaction, and production of BDNF by enzyme-linked immunoassay was determined. Data indicate that, in SH-SY5Y as well as in cortical astrocytes, after 4 hr sodium L-lactate increases the expression and release of BDNF, iNOS, and NO; after 24 hr, it turns is ineffective for the production of the neurotrophin in SH-SY5Y and not in astrocytes, but the expression of iNOS and release of NO appear to be further increased compared with those after 4 hr. Sodium L-lactate influences differently the expression of HSP70 in SH-SY5Y compared with astrocytes. We propose, based on these findings, that sodium L-lactate affects the expression of BDNF in SH-SY5Y and astrocytes in a different manner: high levels of iNOS and NO expressed in SH-SY5Y have a profound inhibitory effect on the release of BDNF related to a more limited production of HSP70 by SH-SY5Y. In conclusion, the results demonstrate differences in the responses of SH-SY5Y and astrocytes to stimulation by high levels of sodium L-lactate. Sodium L-lactate differently and dose and time dependently influences the expression and release of BDNF, iNOS, NO, and HSP70 depending on the cell type.
本研究分析了在 5、10 和 25mmol/L 浓度下添加 L-乳酸钠对人星形胶质细胞和 SH-SY5Y 细胞系的体外影响。通过 Western blot 分析评估脑源性神经营养因子 (BDNF)、诱导型一氧化氮合酶 (iNOS) 和热休克蛋白 70kDa (HSP70) 的表达。通过 MTT 测定细胞活力、通过 Griess 反应测定一氧化氮 (NO) 的释放以及通过酶联免疫吸附测定法测定 BDNF 的产生。数据表明,在 SH-SY5Y 以及皮质星形胶质细胞中,4 小时后 L-乳酸钠增加 BDNF、iNOS 和 NO 的表达和释放;24 小时后,它对 SH-SY5Y 中神经营养因子的产生无效,但 iNOS 的表达和 NO 的释放似乎比 4 小时后进一步增加。与星形胶质细胞相比,L-乳酸钠对 SH-SY5Y 中 HSP70 的表达有不同的影响。基于这些发现,我们提出,L-乳酸钠以不同的方式影响 SH-SY5Y 和星形胶质细胞中 BDNF 的表达:在 SH-SY5Y 中表达的高水平 iNOS 和 NO 对 BDNF 的释放有深远的抑制作用,这与 SH-SY5Y 中 HSP70 的产生更为有限有关。总之,这些结果表明,SH-SY5Y 和星形胶质细胞对高水平 L-乳酸钠刺激的反应存在差异。L-乳酸钠根据细胞类型的不同,以不同的方式和时间依赖性方式影响 BDNF、iNOS、NO 和 HSP70 的表达和释放。