Effect of adenosine on calcium uptake by intact and cultured vascular smooth muscle.

The effect of adenosine (ADO), a potent vasodilator, on the cellular calcium uptake by vascular smooth muscle (VSM) was studied. In addition, similar experiments were conducted with other vasoactive agents to ensure the validity of results obtained with the cultured VSM cell model. Primary VSM monolayers from rat aorta were incubated for 30 min in 45Ca-enriched physiological salt solution (PSS) (37 degrees C) and washed with 20 mM Ca-EGTA (4 degrees C) to remove extracellular 45Ca. Calcium uptake with 4.6 mM K+ was 65.3 +/- 2.4 pmol Ca/10(5) cells. Verapamil (10(-6) M) or 4 degrees C incubation decreased this value 27 and 65%, respectively, whereas ADO (10(-7) M) and isoproterenol (10(-6) M) elevated calcium uptake 21 and 9%, respectively. Elevation of extracellular K+ (25 mM) increased calcium uptake to 87.8 +/- 5.0 pmol Ca/10(5) cells. ADO (10(-7) M), isoproterenol (10(-6) M), and 4 degrees C incubation significantly attenuated this K+-induced increase 25, 19, and 64%, respectively. The calcium uptake of nondepolarized cells was not changed by norepinephrine (10(-6) M) in the presence of either alpha- or beta-adrenergic blockade. Experiments were performed in a similar manner with porcine carotid artery strips in the presence of alpha- and beta-adrenergic blockade. Calcium uptake increased from 2.3 +/- 0.2 X 10(-7) to 3.0 +/- 0.1 X 10(-7) mol Ca/g wet wt after elevation of the medium K+ to 25 mM. This increase was inhibited by 10(-6) M ADO. ADO (10(-6) M) was found to have no effect on the 45Ca efflux from cultured VSM. It is concluded that ADO relaxes VSM by reducing the inward movement of calcium during stimulation. The significant ADO-induced elevation of cellular calcium under conditions when cultured cells were not partially depolarized suggests the existence of an enhanced calcium sequestration within these cells.