Study on pathophysiological roles of brain NO-NCX signal

本研究在HEK 293细胞表达系统中检测了脑特异性NCX剪接变体NCX1.5在NO诱导的细胞毒性中的作用。将含有大鼠NCX1.5全长cDNA的质粒pcDNA3.1/V5-His转染HEK 293细胞，可显著提高NCX1.5 C端V5和（His）6表位（NCX1.5-V5/His）的蛋白水平。硝普钠和S-亚硝基-N-乙酰青霉胺对转染细胞和对照细胞的细胞毒作用无差异。 这些结果表明NO细胞毒性不依赖于NCX1.5。微透析研究表明，NO不影响脑内单胺释放。RT-PCR分析表明，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.