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. 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.

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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

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