Hypoxia-induced cytosolic calcium influx is mediated primarily by the reverse mode of Na+/Ca2+ exchanger in smooth muscle cells of fetal small pulmonary arteries.

AIM: Constriction of small pulmonary arteries and high resistance of pulmonary circulation are important for maintaining fetal circulation before birth. In this study, we investigated how cytosolic free calcium concentration ([Ca(2+)]i) in fetal lamb small pulmonary artery smooth muscle cells (SPASMC) was affected by hypoxia and regulated by calcium pumps during this process. METHODS: (Ca(2+))i in response to acute hypoxia was determined spectrofluorometrically with fluo-3AM in cultured fetal SPASMC. Chemicals or solutions, including ryanodine, 2-aminoethoxydiphenyl borate, Ca(2+)-free solution with 20 mmol ethyleneglycoltetraacetic (EGTA), nimodipine, Na(+)-free medium and KB-R7943, were administrated at the same time point when samples were exposed to acute hypoxia. RESULTS: (Ca(2+))i in fetal lamb SPASMC increased under acute hypoxia. 2-Aminoethoxydiphenyl borate, an inhibitor of inositol triphosphate calcium store, partially attenuated the (Ca(2+))i increase after 6-min treatment. Ryanodine, an inhibitor of ryanodine-sensitive calcium stores, had no effect on the (Ca(2+))i increase. Ca(2+)-free solution with EGTA completely abolished this increase. Both nimodipine, that blocks the voltage-gated calcium channel, and KB-R7943, that inhibits the reverse mode of Na(+)/Ca(2+) exchanger, greatly diminished the hypoxia-induced (Ca(2+))i increase. The inhibitory effect of KB-R7943 was stronger than nimodipine, evidenced by the fact that (Ca(2+))i dropped near to the baseline level in the presence of KB-R7943 at a later time point. Low extracellular Na(+) concentration enhanced the hypoxia-induced increase of (Ca(2+))i. CONCLUSION: These results suggest that hypoxia-induced Ca(2+) increase in fetal SPASMC results from cytosolic Ca(2+) influx mediated primarily by the reverse mode of Na(+)/Ca(2+) exchanger.