Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025 Mar 18:23.
doi: 10.18332/tid/201400. eCollection 2025.

Protective effects of Apelin-13 on nicotine-induced H9c2 cardiomyocyte apoptosis and oxidative stress

Can Xu  1 , Xinyu Nie  2 , Ru Xu  2 , Luyang Zhou  3 , Dongjin Wang  1
Affiliations

Protective effects of Apelin-13 on nicotine-induced H9c2 cardiomyocyte apoptosis and oxidative stress

Can Xu et al. Tob Induc Dis. .

Abstract

Introduction: We aimed to explore the role of Apelin-13 in resisting oxidation, inflammation as well as apoptosis and its underlying mechanisms of action using a model of nicotine-induced H9c2 cardiomyocyte injury.

Methods: H9c2 cardiomyocytes were randomly divided into control, nicotine, nicotine + Apelin-13, and Apelin-13 groups. Cell counting kit-8 assay was conducted to determine the cell viability. Interleukin (IL)-6, superoxide dismutase, tumor necrosis factor-alpha (TNF-α), glutathione peroxidase (GSH-Px), IL-β, catalase (CAT), IL-8, lactate dehydrogenase (LDH), and malondialdehyde (MDA) levels were examined. A 2',7'-dichlorodihydrofluorescein diacetate assay was conducted to measure the intracellular reactive oxygen species (ROS) level. The morphology of apoptotic cardiomyocytes was observed by 4',6-diamidino-2-phenylindole staining. Western blotting was employed to measure the protein expressions of apoptotic factors B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X (Bax). Apoptosis was quantified using Annexin V/propidium iodide staining.

Results: Exposure of H9c2 cardiomyocytes to 10 μM nicotine significantly reduced cell viability and increased LDH release, oxidative stress (elevated MDA and ROS levels with decreased superoxide dismutase, GSH-Px, and CAT activities), pro-inflammatory cytokines (IL-6, TNF-α, IL-1β, IL-8), and apoptotic markers (increased Bax with decreased Bcl-2 expression, along with nuclear condensation) (p<0.05). In contrast, treatment with 2 μM Apelin-13 significantly alleviated these deleterious effects, enhancing cell viability, restoring antioxidant enzyme activities, reducing oxidative and inflammatory responses, and inhibiting apoptosis (p<0.05).

Conclusions: Nicotine induction increases the oxidative stress and apoptotic capacity of H9c2 cardiomyocytes, but Apelin-13 protects H9c2 cardiomyocytes against nicotine-induced apoptosis and oxidative stress.

Keywords: apelin-13; apoptosis; cardiomyocyte; nicotine; oxidative stress.

PubMed Disclaimer

Conflict of interest statement

The authors have completed and submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest and none was reported.

Figures

Figure 1
Figure 1
Effect of Apelin-13 on viability of H9c2 cardiomyocytes treated with nicotine: A and B) Toxicity of nicotine at various concentrations towards H9c2 cardiomyocytes; C and D) effects of Apelin-13 at various concentrations on H9c2 cardiomyocytes treated with 10 μM nicotine; E) LDH activity assay results
Figure 2
Figure 2
Effect of Apelin-13 on: A) SOD activity, B) MDA level, C) CAT activity and D) GSH-Px activity in H9c2 cardiomyocytes treated with nicotine
Figure 3
Figure 3
Effect of Apelin-13 on ROS production in H9c2 cardiomyocytes treated with nicotine: A) DCFH-DA assay results; B and C) ROS levels
Figure 4
Figure 4
Effect of Apelin-13 on: A) TNF-α, B) IL-6, C) IL-β and D) IL-8 levels in H9c2 cardiomyocytes
Figure 5
Figure 5
Effect of Apelin-13 on the morphology of H9c2 cardiomyocytes treated with nicotine
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Saour B, Smith B, Yancy CW. Heart failure and sudden cardiac death. Card Electrophysiol Clin. 2017;9(4):709-723. doi:10.1016/j.ccep.2017.07.010 - DOI - PubMed
    1. Naito R, Miyauchi K. Coronary artery disease and type 2 diabetes mellitus. Int Heart J. 2017;58(4):475-480. doi:10.1536/ihj.17-191 - DOI - PubMed
    1. Kaneto CM, Nascimento JS, Prado MSJG, Mendonça LSO. Circulating miRNAs as biomarkers in cardiovascular diseases. Eur Rev Med Pharmacol Sci. 2019;23(5):2234-2243. doi:10.26355/eurrev_201903_17271 - DOI - PubMed
    1. Tao J, Chen BD, Ma YT, et al. . FrzA gene protects cardiomyocytes from H2O2-induced oxidative stress through restraining the Wnt/Frizzled pathway. Lipids Health Dis. 2015;14:90. doi:10.1186/s12944-015-0088-0 - DOI - PMC - PubMed
    1. Heistad DD, Wakisaka Y, Miller J, Chu Y, Pena-Silva R. Novel aspects of oxidative stress in cardiovascular diseases. Circ J. 2009;73(2):201-207. doi:10.1253/circj.cj-08-1082 - DOI - PMC - PubMed

LinkOut - more resources