Magnesium-assisted hydrogen improves isoproterenol-induced heart failure

Chen, Fengbao1,#; Chen, Ruimin1,#; Yang, Lili1,2; Shen, Bowen1,3; Wang, Yunting1; Gao, Yongfeng1; Tan, Rui1,*; Zhao, Xiaomin1,*

Author Information

1Institute of Pharmacology, Shandong First Medical University & Shandong Academy of Medical Sciences, Taian, Shandong Province, China

2New Drug Evaluation Center of Shandong Academy of Pharmaceutical Sciences, Shandong Academy of Pharmaceutical Sciences, Ji’nan, Shandong Province, China

3School of Chemistry and Pharmaceutical Engineering, Shandong First Medical University & Shandong Academy of Medical Sciences, Taian, Shandong Province, China

*Correspondence to: Xiaomin Zhao, PhD, xmzhao@sdfmu.edu.cn; Rui Tan, PhD, rtan@sdfmu.edu.cn.

#Both authors contributed equally to this work.

Funding:This study was supported by the Pharmacology Key Discipline Foundation of Shandong Province (No. ZDXK00001) and the Collaborative Innovation Center for Research and Development of Traditional Chinese Medicine in Mount Tai (No. zd099).

Abstract

Heart failure (HF) is a leading cause of mortality among patients with cardiovascular disease and is often associated with myocardial apoptosis and endoplasmic reticulum stress (ERS). While hydrogen has demonstrated potential in reducing oxidative stress and ERS, recent evidence suggests that magnesium may aid in hydrogen release within the body, further enhancing these protective effects. This study aimed to investigate the cardioprotective effects of magnesium in reducing apoptosis and ERS through hydrogen release in a rat model of isoproterenol (ISO)-induced HF. Magnesium was administered orally to ISO-induced HF rats, which improved cardiac function, reduced myocardial fibrosis and cardiac hypertrophy, and lowered the plasma levels of creatine kinase-MB, cardiac troponin-I, and N-terminal B-type natriuretic peptide precursor in ISO-induced HF rats. It also inhibited cardiomyocyte apoptosis by upregulating B-cell lymphoma-2, downregulating Bcl-2-associated X protein, and suppressing ERS markers (glucose-related protein 78, activating transcription factor 4, and C/EBP-homologous protein). Magnesium also elevated hydrogen levels in blood, plasma, and cardiac tissue, as well as in artificial gastric juice and pure water, where hydrogen release lasted for at least four hours. Additionally, complementary in vitro experiments were conducted using H9C2 cardiomyocyte injury models, with hydrogen-rich culture medium as the intervention. Hydrogen-rich culture medium improved the survival and proliferation of ISO-treated H9C2 cells, reduced the cell surface area, inhibited apoptosis, and downregulated ERS pathway proteins. However, the protective effects of hydrogen were negated by tunicamycin (an inducer of ERS) in H9C2 cells. In conclusion, magnesium exerts significant cardioprotection by mitigating ERS and apoptosis through hydrogen release effects in ISO-induced HF.

摘要

心力衰竭（HF）是心血管疾病患者死亡的主要原因，通常与心肌细胞凋亡和内质网应激（ERS）相关。虽然氢已被证明具有减轻氧化应激和ERS的潜力，但近期研究表明，镁可能有助于氢在体内的释放，从而进一步增强这些保护作用。本研究旨在探讨在异丙肾上腺素（ISO）诱导的心力衰竭大鼠模型中，镁通过促进氢释放以减少细胞凋亡和ERS从而发挥心脏保护作用的机制。对ISO诱导的心力衰竭大鼠给予镁剂口服后，结果显示其心功能得到改善，心肌纤维化和心脏肥大减轻，血浆中肌酸激酶同工酶、心肌肌钙蛋白I 和N末端B型利钠肽前体水平降低。同时，镁通过上调B细胞淋巴瘤-2、下调Bcl-2相关X蛋白抑制了心肌细胞凋亡，并抑制了ERS标志物（葡萄糖调节蛋白78、激活转录因子4 和C/EBP同源蛋白）的表达。镁还提高了血液、血浆、心脏组织以及人工胃液和纯水中的氢水平，且氢的释放可持续至少四小时。此外，本研究还利用H9C2心肌细胞损伤模型进行了补充性体外实验，并以富氢培养基作为干预手段。富氢培养基改善了ISO处理的H9C2细胞的存活与增殖，减小了细胞表面积，抑制了细胞凋亡，并下调了ERS通路蛋白的表达。然而，在H9C2细胞中，氢的保护作用被衣霉素（一种ERS诱导剂）所抵消。综上所述，在ISO诱导的心力衰竭模型中，镁通过其氢释放效应减轻内质网应激和细胞凋亡，从而发挥显著的心脏保护作用。