Functional coupling of angiotensin II type 1 receptor with insulin resistance of energy substrate uptakes in immortalized cardiomyocytes (HL‐1 cells)

Background and purpose: Increased angiotensin II levels and insulin resistance coexist at the early stages of cardiomyopathies. To determine whether angiotensin II increases insulin resistance in cardiomyocytes, we studied the effect of angiotensin II on basal and insulin‐stimulated transport rate of energy substrates in immortalized cardiomyocytes (HL‐1 cells). Experimental approach: Glucose and palmitic acid uptakes were measured using [ 3 H]2‐deoxy‐ D ‐glucose and [ 14 C]palmitic acid, respectively, in cells exposed or not exposed to angiotensin II (100 n M ), angiotensin II plus irbesartan or PD123319, type 1 and 2 receptor antagonists, or PD98059, an inhibitor of ERK1/2 activation. Cell viability, DNA, protein synthesis and surface area were evaluated by the MTT test, [ 3 H]thymydine, [ 3 H]leucine and morphometric analysis, respectively. Type 1 receptor levels were measured by western blot analysis. Key results: Basal uptakes of glucose and palmitic acid by HL‐1 cells (0.37±0.07 and 7.31±0.22 pmol per 10 4 cells per min, respectively) were both stimulated by 100 n M insulin (+91 and +64%, respectively). Cells exposed to angiotensin II remained viable and did not show signs of hypertrophy. In these conditions, the basal palmitic acid uptake of the cells increased (11.41±0.46 pmol per 10 4 cells per min) and insulin failed to stimulate the uptake of glucose and fatty acids. Changes in the rate of uptake of energy substrates were prevented or significantly reduced by irbesartan or PD98059. Conclusions and implications: Angiotensin II is a candidate for increasing insulin resistance in cardiomyocytes. Our results suggest a further mechanism for the cardiovascular protection offered by the angiotensin II type 1 receptor blockers. British Journal of Pharmacology (2008) 153 , 907–914; doi: 10.1038/sj.bjp.0707563 ; published online 5 November 2007