AT₁-receptor response to non-saturating Ang-II concentrations is amplified by calcium channel blockers

Blockers of angiotensin II type 1 receptor (AT₁R) and the voltage gated calcium channel 1.2 (Ca_V1.2) are commonly used for treatment of hypertension. Yet there is little information about the effect of physiological concentrations of angiotensin II (AngII) on AT₁R signaling and whether there is a reciprocal regulation of AT₁R signaling by Ca_V1.2.

To elucidate these questions, we have studied the Ca²⁺ signaling response to physiological and pharmacological AngII doses in HEK293a cells, vascular smooth muscle cells and cardiomyocytes using a Ca²⁺ sensitive dye as the principal sensor. Intra-cellular calcium recordings were performed in presence and absence of Ca_V1.2 blockers. Semi-quantitative imaging methods were used to assess the plasma membrane expression of AT₁R and G-protein activation.

Repeated exposure to pharmacological (100 nM) concentrations of AngII caused, as expected, a down-regulation of the Ca²⁺ response. In contrast, repeated exposure to physiological (1 nM) AngII concentration resulted in an enhancement of the Ca²⁺ response. The up-regulation of the Ca²⁺ response to repeated 1 nM AngII doses and the down-regulation of the Ca²⁺ response to repeated 100 nM Angll doses were not accompanied by a parallel change of the AT₁R plasma membrane expression. The Ca²⁺ response to 1 nM of AngII was amplified in the presence of therapeutic concentrations of the Ca_V1.2 blockers, nifedipine and verapamil, in vascular smooth muscle cells, cardiomyocytes and HEK293a cells. Amplification of the AT₁R response was also observed following inhibition of the calcium permeable transient receptor potential cation channels, suggesting that the activity of AT₁R is sensitive to calcium influx.

Our findings have implications for the understanding of hyperactivity of the angiotensin system and for use of Ca²⁺ channel blockers as mono-therapy in hypertension.