Verónica Rebollar González

Simultaneous Rho kinase inhibition in circulating leukocytes and in cardiovascular tissue in rats with high angiotensin converting enzyme levels

BACKGROUND The small guanosine triphosphatase RhoA and its direct target Rho kinase (ROCK) play important roles in cardiovascular pathophysiology. Activated ROCK phosphorylates intracellular proteins with detrimental effects on cardiovascular remodeling. Increased ROCK activity in circulating leukocytes is observed in hypertension and in heart failure, but its relationship with ROCK activation in the myocardium and vessels is unknown. We hypothesized that ROCK activation and phosphorylation/activation of some of its key downstream molecules in the heart and arterial wall are reflected in circulating leukocytes. METHODS Phosphorylation of MYPT1, ERM and p38-MAPK and levels of p65-NF-κB were determined in the left ventricle (LV), aortic wall and circulating leukocytes in rats with high (Brown Norway, BN) and low (Lewis) angiotensin converting enzyme. A group of BN rats received the ROCK inhibitor fasudil (7days). RESULTS Compared to Lewis rats, in the BN group phosphorylated levels of MYPT1, ERM and p38-MAPK and levels of p65-NF-κB were increased (P<0.05) in the LV (67%, 92%, 52% and 98%, respectively); in the aortic wall (57%, 51%, 68% and 66%, respectively) and in circulating leukocytes (61%, 72%, 49% and 105%, respectively). Fasudil reduced all these levels to those observed in Lewis rats. Phosphorylated MYPT1, ERM, and p38-MAPK and levels of p65-NF-κB in circulating leukocytes were significantly correlated with their respective LV and aortic wall levels (excepting p65-NF-κB in aorta). CONCLUSION ROCK activity in circulating leukocytes reflects activation of this signaling pathway in the myocardium and aortic wall in this model, and supports its value as a potential cardiovascular remodeling marker.