Study on pathophysiological roles of brain NO-NCX signal

本研究在HEK293细胞表达系统中检测了主要脑特异性NCX剪接变体NCX1.5在no诱导的细胞毒性中的作用。用含有全长大鼠NCX1.5 cDNA的质粒构建体pcDNA3.1/V5-His转染HEK293细胞后，c端V5和（His）6表位标记的NCX1.5 （NCX1.5-V5/His）蛋白水平显著升高。硝普钠和s -亚硝基-n -乙酰青霉胺的细胞毒作用在对照细胞和转染细胞之间没有差异。这些结果表明NO的细胞毒性不依赖于NCX1.5。微透析研究表明NO不影响脑内单胺的释放。此外，RT-PCR分析显示，环境因素对NCX的表达没有影响。

This study examined the role of the predominant brain-specific NCX splice variant NCX1.5 in NO-induced cytotoxicity in HEK293 cell expression system. Transfection with a plasmid construct pcDNA3.1/V5-His containing full-length rat NCX1.5 cDNA into HEK293 cells increased prominently the protein level of NCX1.5 tagged with C-terminal V5 and (His)6 epitopes (NCX1.5-V5/His). There was no difference in the cytotoxic effects by sodium nitroprusside and S-nitroso-N-acetylpenicillamine between the control and transfected cells. These results suggest that NO cytotoxicity is not dependent on NCX1.5. Microdialysis study showed that NO did not affect monoamine release in the brain. Furthermore, RT-PCR analysis showed that environmental factors did not affect the expression of NCX.