ANGIOTENSIN-II RESPONSES AFTER PROTEIN-KINASE-C ACTIVATION IN VASCULAR SMOOTH-MUSCLE CELLS OF SPONTANEOUSLY HYPERTENSIVE RATS

To examine the interaction of protein kinase C (PKC) with agonist-indu ced calcium fluxes in hypertension, cytosolic free calcium ([Ca2+]i) w as measured in vascular smooth muscle cells (vSMC) of normotensive and spontaneously hyper-tensive rats (SHR) after incubation with phorbol, -12 myristate,-13 acetate (PMA) and application of angiotensin II (AII ). To distinguish between calcium influx through voltage-dependent cal cium channels and calcium mobilization from intracellular stores, the calcium agonist BayK 8644 was used. Resting [Ca2+]i was 108.0 +/- 10.6 nM (mean +/- SEM, n = 25) in normotensive and 102.0 +/- 11.4 nM (n = 21) in hypertensive cells. After pretreatment with PMA 10(-7) M for 60 min, resting [Ca2+]i of normotensive vSMC increased to 145.0 +/- 13.8 nM (n = 17) while the resting level of the hyper-tensive cells decrea sed to 68.0 +/- 2.4 nM (n = 14, p < 0.05 as compared with normotensive cells) in hypertensive vSMC. Maximum increase in [Ca2+]i induced with 10 M AII for normotensive and hypertensive vSMC was similar: 230.5 +/ - 34.4 nM (n = 14) and 212.5 +/- 26.7 nM (n = 17). After pretreatment with PMA 10(-7) M, the maximum increase in [Ca2+]i induced by AII in h ypertensive cells was limited to 108.0 +/- 6.2 nM (p < 0.05 as compare d with normotensive cells), whereas the increase in [Ca2+]i in normote nsive vSMC remained the same as before: 211.5 +/- 23.4 nM. After admin istration of 10(-5) M BayK 8644, [Ca2+]i increased by 54.3 +/- 12.2 nM (n = 4) and 43.4 +/- 17.4 nM (n = 5) in normotensive and hypertensive vSMC, respectively. After preincubation with PMA this effect was redu ced to 20.3 +/- 8.8 nM in normotensive cells and 29.0 +/- 4.6 nM in hy pertensive cells. We conclude that in hypertensive vSMC either the fee dback inhibition of postreceptor signaling after PKC activation is mor e pronounced or that higher amounts of PKC can be activated.