Hydrogen (H2) Inhibits Isoproterenol-Induced Cardiac Hypertrophy via Antioxidative Pathways. | イソプロテレノールによる心肥大を 水素は抗酸化経路を介して阻害する

Front Pharmacol. 2016 Oct 27;7:392. eCollection 2016.

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

Department of Physiology, Zhongshan School of Medicine, Sun Yat-sen University


Background and Purpose:

Hydrogen (H2) 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 H2 in a model of isoproterenol (ISO)-induced cardiac hypertrophy.


Mice (C57BL/6J; 8-10 weeks of age) were randomly assigned to four groups: Control group (n = 10), ISO group (n = 12), ISO plus H2 group (n = 12), and H2 group (n = 12). Mice received H2 (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 H2 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 H2-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.


Intraperitoneal injection of H2 prevented cardiac hypertrophy and improved cardiac function in ISO-infused mice. H2-rich medium blocked ISO-mediated cardiomyocytes hypertrophy in vitro. H2 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.


H2 inhibits ISO-induced cardiac/cardiomyocytes hypertrophy both in vivo and in vitro, and improves the impaired left ventricular function. H2 exerts its protective effects partially through blocking ROS-sensitive ERK1/2, p38, and JNK signaling pathways.

Front Pharmacol. 2016 Oct 27;7:392. eCollection 2016.


中山大学 中山医学院 生理学科







一方in vitro試験では、H9c2心筋細胞を高濃度の水素培養液で30分間事前に処理し、その後ISO(10μM)を負荷しました。その後、培養液とISOは24時間おきに入れ替えました。

心筋細胞の表層部位と、ANP/BNP相対的mRNA量、3-ニトロチロシン発現量、ミトコンドリア膜電位(MMP)消失についても調べました。さらに細胞外ERK1/2、p-ERK1/2、p38、p-p38、c-JNK、p-JNKをin vivoとin vitroそれぞれで測定しました。


水素を腹腔内注射することにより、ISO投与マウスの心肥大が抑えられ、心機能が改善しました。また、in vitroでも高濃度水素培養液を用いると、ISO添加による心筋細胞の容積増大が抑えられました。そして、水素はNADPH酸化酵素の過剰発現とROS蓄積を抑え、MMPの減少を軽減し、ROS感受性ERK1/2、p38、JNKシグナル経路を阻害しました。


水素は in vivoまたはin vitroの両方で、心肥大および心筋細胞の容積増大を抑え、低下した左心室の機能を改善しました。そしてその水素による保護作用の少なくとも一部は、ROS感受性ERK1/2、p38、JNLシグナル経路の遮断を介することで発揮されます。