Interaction of the Na +-Ca 2+ exchanger with small molecules on cell Ca 2+ signaling

Abstract

Interactions of the Na(+)-Ca2+ exchanger with small molecules on cell Ca2+ signaling were elucidated in Chinese hamster ovary (CHO) C1 cells, which transfected a control vector without any expression of the Na(+)-Ca2+ exchanger's gene while CHO CK1.4 cells transfected an expression vector encoding the bovine cardiac Na(+)-Ca2+ exchanger's cDNA, treated with lithium- or sodium-buffer medium respectively, by using L16(2)15 multifactorial orthogonal statistics and fura-2 fluorescence real-time imaging. In contrast to controls of Li(+)-treated C1 cells, the store-dependent Ca(2+)-influx (SDCI) was enhanced by either the Na(+)-Ca2+ exchanger, Na(+), 1-¿(beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl¿-1H-imidazole HCl (SK&F96365) or ouabain, and by interactions of the Na(+)-Ca2+ exchanger with either Na+, SK&F96365 or both SK&F96365 and ouabain; and ATP-induced Ca2+ release (AICR) was activated by SK&F96365 or Na+ alone, interactions of the Na(+)-Ca2+ exchanger with SK&F96365 or Na+, and an interaction between SK&F96365 and ouabain. The dramatic interaction of the Na(+)-Ca2+ exchanger with small molecules indicates that cell Ca2+ signaling is generated by inositol triphosphate (InsP3)-dependent pathways, allosteric effects of the G-protein coupled P2y&2u purinoceptor and multi-site recognition. Our findings provide meaningful clues for designing new strategies of cardiocerebral vascular oxidative diseases.