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Background and Purpose:  Hydrogen (H 2 ) has been shown to have a strong antioxidant effect on preventing oxidative stress-related diseases. The goal of the present study is to determine the pharmacodynamics of H 2  in a model of isoproterenol (ISO)-induced cardiac hypertrophy.   Methods:  Mice (C57BL/6J; 8–10 weeks of age) were randomly assigned to four groups: Control group ( n  = 10), ISO group ( n  = 12), ISO plus H 2  group ( n  = 12), and H 2  group ( n  = 12). Mice received H 2  (1 ml/100g/day, intraperitoneal injection) for 7 days before ISO (0.5 mg/100g/day, subcutaneous injection) infusion, and then received ISO with or without H 2  for another 7 days. Then, cardiac function was evaluated by echocardiography. Cardiac hypertrophy was reflected by heart weight/body weight, gross morphology of hearts, and heart sections stained with hematoxylin and eosin, and relative atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) mRNA levels. Cardiac reactive oxygen species (ROS), 3-nitrotyrosine and p67 (phox) levels were analyzed by dihydroethidium staining, immunohistochemistry and Western blotting, respectively. For  in vitro  study, H9c2 cardiomyocytes were pretreated with H 2 -rich medium for 30 min, and then treated with ISO (10 μM) for the indicated time. The medium and ISO were re-changed every 24 h. Cardiomyocyte surface areas, relative ANP and BNP mRNA levels, the expression of 3-nitrotyrosine, and the dissipation of mitochondrial membrane potential (MMP) were examined. Moreover, the expression of extracellular signal-regulated kinase1/2 (ERK1/2), p-ERK1/2, p38, p-p38, c-Jun NH2-terminal kinase (JNK), and p-JNK were measured by Western blotting both  in vivo  and  in vitro .   Results:  Intraperitoneal injection of H 2  prevented cardiac hypertrophy and improved cardiac function in ISO-infused mice. H 2 -rich medium blocked ISO-mediated cardiomyocytes hypertrophy  in vitro.  H 2  blocked the excessive expression of NADPH oxidase and the accumulation of ROS, attenuated the decrease of MMP, and inhibited ROS-sensitive ERK1/2, p38, and JNK signaling pathways.   Conclusion:  H 2  inhibits ISO-induced cardiac/cardiomyocytes hypertrophy both  in vivo  and  in vitro , and improves the impaired left ventricular function. H 2  exerts its protective effects partially through blocking ROS-sensitive ERK1/2, p38, and JNK signaling pathways.

Hydrogen (H2) Inhibits Isoproterenol-Induced Cardiac Hypertrophy via Antioxidative Pathways