Homocysteine induces cardiomyocyte dysfunction and apoptosis through p38 MAPK-mediated increase in oxidant stress

Abstract 

Elevated plasma homocysteine (Hcy) is a risk factor for cardiovascular disease. While Hcy has been shown to promote endothelial dysfunction by decreasing the bioavailability of nitric oxide and increasing oxidative stress in the vasculature, the effects of Hcy on cardiomyocytes remain less understood. In this study we explored the effects of hyperhomocysteinemia (HHcy) on myocardial function ex vivo and examined the direct effects of Hcy on cardiomyocyte function and survival in vitro. Studies with isolated hearts from wild type and HHcy mice (heterozygous cystathionine-beta synthase deficient mice) demonstrated that HHcy mouse hearts had more severely impaired cardiac relaxation and contractile function and increased cell death following ischemia reperfusion (I/R). In isolated cultured adult rat ventricular myocytes, exposure to Hcy for 24h impaired cardiomyocyte contractility in a concentration-dependent manner, and promoted apoptosis as revealed by terminal dUTP nick-end labeling and cleaved caspase-3 immunoblotting. These effects were associated with activation of p38 MAPK, decreased expression of thioredoxin (TRX) protein, and increased production of reactive oxygen species (ROS). Inhibition of p38 MAPK by the selective inhibitor SB203580 (5μM) prevented all of these Hcy-induced changes. Furthermore, adenovirus-mediated overexpression of TRX in cardiomyocytes significantly attenuated Hcy-induced ROS generation, apoptosis, and impairment of myocyte contractility. Thus, Hcy may increase the risk for CVD not only by causing endothelial dysfunction, but also by directly exerting detrimental effects on cardiomyocytes.

Abbreviations: Hcy, homocysteine, HHcy, hyperhomocysteinemia, tHcy, total homocysteine, TRX, thioredoxin, CBS, cystathionine β-synthase, CVD, cardiovascular disease, NO, nitric oxide, ROS, reactive oxygen species, % CS, percent cell shortening, [Ca²⁺]_i, intracellular calcium, WT, wild type, TTE, trans-thoracic echocardiography, EDP, end diastolic pressure, ESP, end systolic pressure, I/R, ischemia reperfusion, TUNEL, terminal dUTP nick-end labeling

Keywords: Homocysteine, Hyperhomocysteinemia, Cardiomyocytes, Contractility, Apoptosis, Antioxidant, Ischemia reperfusion injury, Oxidative stress