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J Ginseng Res. 2020 Mar;44(2):258-266. doi: 10.1016/j.jgr.2018.12.004. Epub 2018 Dec 16.

The role of ginsenoside Rb1, a potential natural glutathione reductase agonist, in preventing oxidative stress-induced apoptosis of H9C2 cells.

Author information

1
School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, China.
2
Department of Traditional Chinese Medicine, The first hospital of Yangjiang, Yangjiang, China.
3
Department of Orthopaedics and Traumatology, Queen Mary Hospital, the University of Hong Kong, Hong Kong.
4
Department of Cardiology, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
5
Department of Traditional Chinese Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, China.

Abstract

Background:

Oxidative stress-induced cardiomyocytes apoptosis is a key pathological process in ischemic heart disease. Glutathione reductase (GR) reduces glutathione disulfide to glutathione (GSH) to alleviate oxidative stress. Ginsenoside Rb1 (GRb1) prevents the apoptosis of cardiomyocytes; however, the role of GR in this process is unclear. Therefore, the effects of GRb1 on GR were investigated in this study.

Methods:

The antiapoptotic effects of GRb1 were evaluated in H9C2 cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, annexin V/propidium iodide staining, and Western blotting. The antioxidative effects were measured by a reactive oxygen species assay, and GSH levels and GR activity were examined in the presence and absence of the GR inhibitor 1,3-bis-(2-chloroethyl)-1-nitrosourea. Molecular docking and molecular dynamics simulations were used to investigate the binding of GRb1 to GR. The direct influence of GRb1 on GR was confirmed by recombinant human GR protein.

Results:

GRb1 pretreatment caused dose-dependent inhibition of tert-butyl hydroperoxide-induced cell apoptosis, at a level comparable to that of the positive control N-acetyl-L-cysteine. The binding energy between GRb1 and GR was positive (-6.426 kcal/mol), and the binding was stable. GRb1 significantly reduced reactive oxygen species production and increased GSH level and GR activity without altering GR protein expression in H9C2 cells. Moreover, GRb1 enhanced the recombinant human GR protein activity in vitro, with a half-maximal effective concentration of ≈2.317 μM. Conversely, 1,3-bis-(2-chloroethyl)-1-nitrosourea co-treatment significantly abolished the GRb1's apoptotic and antioxidative effects of GRb1 in H9C2 cells.

Conclusion:

GRb1 is a potential natural GR agonist that protects against oxidative stress-induced apoptosis of H9C2 cells.

KEYWORDS:

Apoptosis; Ginsenoside Rb1; Glutathione reductase; H9C2; Oxidative stress

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