Hydrogen inhibits isoproterenol‑induced autophagy in cardiomyocytes in vitro and in vivo.
Hydrogen inhibits isoproterenol‑induced autophagy in cardiomyocytes in vitro and in vivo.
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DOI：
10.3892/mmr.2017.7601
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发表时间：
2017-12
影响因子：
3.4
通讯作者：
Yaxing Zhang;Zhiyuan Long;Jingting Xu;Sihua Tan;N. Zhang;Anfei Li;Ling Wang;T. Wang
中科院分区：
文献类型：
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作者：
Yaxing Zhang;Zhiyuan Long;Jingting Xu;Sihua Tan;N. Zhang;Anfei Li;Ling Wang;T. Wang
我们小组先前的一项研究表明，氢给药可以通过靶向活性氧依赖性丝裂原活化蛋白激酶信号转导来减轻体内心血管肥大。本研究的目的是在体内和体外确定氢对β肾上腺素受体激活过程中心肌细胞自噬的影响。我们制备了富氢介质，并使用MB-Pt试剂法测量了氢的浓度。对于体外研究，H9 c2心肌细胞用异丙肾上腺素（ISO; 10 µM）刺激5、15和30 min，然后通过蛋白质印迹法检测自噬标志物微管相关蛋白1轻链3β II（LC 3B II）的蛋白质表达水平。然后通过用富氢培养基预处理H9 c2心肌细胞30分钟，然后用ISO刺激，并检查自噬标记物LC 3B II的蛋白质表达水平来测试富氢培养基的作用。对于体内研究，小鼠在ISO给药（0.5 mg/100 g/天，通过皮下注射）之前接受氢气（1 ml/100 g/天，通过腹膜内注射）7天，随后在有或没有ISO的情况下接受氢气另外7天。通过心脏重量（HW）和心脏重量/体重（HW/BW）测量检查肥大反应。检测了自噬标志物Beclin 1、自噬相关蛋白7（Atg 7）和LC 3B II的蛋白表达。结果表明，在体外ISO刺激5 min后发生过度的自噬。这种增强的自噬通过用富氢培养基预处理来阻断。此外，氢改善恶化的肥大反应，并抑制增强的自噬活性介导的ISO管理在体内，如通过降低HW和HW/BW，并抑制Beclin 1，Atg 7和LC 3B II的蛋白质表达水平。因此，本研究的结果表明，在体外和体内心肌细胞肥大模型中，氢抑制ISO诱导的过度自噬。
A previous study from our group has demonstrated that hydrogen administration can attenuate cardiovascular hypertrophy in vivo by targeting reactive oxygen species‑dependent mitogen‑activated protein kinase signaling. The aim of the present study is to determine the effect of hydrogen on cardiomyocyte autophagy during β‑adrenoceptor activation in vivo and in vitro. We prepared hydrogen‑rich medium, and the concentration of hydrogen was measured by using the MB‑Pt reagent method. For the in vitro study, H9c2 cardiomyocytes were stimulated with isoproterenol (ISO; 10 µM) for 5, 15 and 30 min, and then the protein expression levels of the autophagy marker microtubule‑associated protein 1 light chain 3β II (LC3B II) were examined by western blotting. The effect of hydrogen‑rich medium was then tested by pretreating the H9c2 cardiomyocytes with hydrogen‑rich medium for 30 min, then stimulating with ISO, and examining the protein expression levels of the autophagy marker LC3B II. For the in vivo study, mice received hydrogen (1 ml/100 g/day, by intraperitoneal injection) for 7 days prior to ISO administration (0.5 mg/100 g/day, by subcutaneous injection), and subsequently received hydrogen with or without ISO for another 7 days. Hypertrophic responses were examined by heart weight (HW) and heart weight/body weight (HW/BW) measurements. The protein expression of autophagy markers Beclin1, autophagy‑related protein 7 (Atg7) and LC3B II were examined. The results demonstrated that excessive autophagy occurred following 5 min of ISO stimulation in vitro. This enhanced autophagy was blocked by pretreatment with hydrogen‑rich medium. Furthermore, hydrogen improved the deteriorated hypertrophic responses and inhibited the enhanced autophagic activity mediated by ISO administration in vivo, as indicated by decreasing HW and HW/BW, and suppressing the protein expression levels of Beclin1, Atg7 and LC3B II. Therefore, the results of the present study demonstrated that hydrogen inhibited ISO‑induced excessive autophagy in cardiomyocyte hypertrophy models in vitro and in vivo.