Angiotensin II Directly Increases Endothelial Calcium and Nitric Oxide in Kidney and Brain Microvessels In Vivo With Reduced Efficacy in Hypertension

Abstract

Background The vasoconstrictor effects of angiotensin II via type 1 angiotensin II receptors in vascular smooth muscle cells are well established, but the direct effects of angiotensin II on vascular endothelial cells (VECs) in vivo and the mechanisms how VECs may mitigate angiotensin II–mediated vasoconstriction are not fully understood. The present study aimed to explore the molecular mechanisms and pathophysiological relevance of the direct actions of angiotensin II on VECs in kidney and brain microvessels in vivo. Methods and Results Changes in VEC intracellular calcium ([Ca2+]i) and nitric oxide (NO) production were visualized by intravital multiphoton microscopy of cadherin 5–Salsa6f mice or the endothelial uptake of NO‐sensitive dye 4‐amino‐5‐methylamino‐2′,7′‐difluorofluorescein diacetate, respectively. Kidney fibrosis by unilateral ureteral obstruction and Ready‐to‐use adeno‐associated virus expressing Mouse Renin 1 gene (Ren1‐AAV) hypertension were used as disease models. Acute systemic angiotensin II injections triggered >4‐fold increases in VEC [Ca2+]i in brain and kidney resistance arterioles and capillaries that were blocked by pretreatment with the type 1 angiotensin II receptor inhibitor losartan, but not by the type 2 angiotensin II receptor inhibitor PD123319. VEC responded to acute angiotensin II by increased NO production as indicated by >1.5‐fold increase in 4‐amino‐5‐methylamino‐2′,7′‐difluorofluorescein diacetate fluorescence intensity. In mice with kidney fibrosis or hypertension, the angiotensin II–induced VEC [Ca2+]i and NO responses were significantly reduced, which was associated with more robust vasoconstrictions, VEC shedding, and microthrombi formation. Conclusions The present study directly visualized angiotensin II–induced increases in VEC [Ca2+]i and NO production that serve to counterbalance agonist‐induced vasoconstriction and maintain residual organ blood flow. These direct and endothelium‐specific angiotensin II effects were blunted in disease conditions and linked to endothelial dysfunction and the development of vascular pathologies.

Keywords

• DAF‐FM
• GCaMP6
• sex specificity
• hypertension
• losartan
• PD123319