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. 2024 Sep-Oct;38(5):2126-2133.
doi: 10.21873/invivo.13675.

The Effect of Hypoxia on Irisin Expression in HL-1 Cardiomyocytes

Maciej Grzeszczuk  1 , Monika Mrozowska  2 , Alicja Kmiecik  2 , Agnieszka Rusak  2 , Karolina Jabłońska  2 , Urszula Ciesielska  2 , Piotr Dzięgiel  2   3 , Katarzyna Nowińska  2
Affiliations

The Effect of Hypoxia on Irisin Expression in HL-1 Cardiomyocytes

Maciej Grzeszczuk et al. In Vivo. 2024 Sep-Oct.

Abstract

Background/aim: Cardiovascular diseases (CVD) are the leading cause of death worldwide. In 2019, 523 million people were diagnosed with CVD, with 18.6 million deaths. Improved treatment and diagnostics could reduce CVD's impact. Irisin (Ir) is crucial for heart function and may be a biomarker for heart attack. Ir is a glycoprotein with sugar residues attached to its protein structure. This glycosylation affects Ir stability, solubility, and receptor interactions on target cells. Its secondary structure includes a fibronectin type III domain, essential for its biological functions. Ir helps cardiomyocytes to respond to hypoxia and protects mitochondria. The aim of the study was to determine the FNDC5 gene expression level and the Ir level in HL-1 cardiomyocytes subjected to hypoxia.

Materials and methods: We examined the effect of hypoxia on the expression levels of the FNDC5 gene and those of Ir in mouse cardiomyocytes of the HL-1 cell line. Real-time PCR (RT-PCR) was used to estimate the expression levels of the FNDC5 gene. Western blot and immunofluorescence methods were used to analyze the Ir protein levels.

Results: Analyses showed an increased Ir level in HL-1 cardiomyocytes in response to hypoxia. This is the first study to confirm the presence of Ir in HL-1 cells.

Conclusion: The observed increase in Ir expression in murine cardiomyocytes is associated with the hypoxic environment and can be potentially used to diagnose hypoxia and CVD.

Keywords: Cardiovascular disease; FNDC5; HIF1α; PGC1α; hypoxia; irisin.

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Conflict of interest statement

The Authors declare no conflicts of interest in relation to this study.

Figures

Figure 1
Figure 1. Comparison of the Ir expression levels by immunofluorescence in mouse cardiomyocytes of HL-1 cells line cultured in normoxic and hypoxic conditions (p<0.0001). Ir: Irisin.
Figure 2
Figure 2. Comparison of Ir and Hif1α expression by confocal microscopy in the HL-1 cells subjected to hypoxia and normoxia. (A) Normoxia - low expression of Ir, magnification 60×. (B) Hypoxia - high expression of Ir, magnification 60×. (C) Normoxia - absence of nuclear expression of Hif1α, magnification 60×. (D) Hypoxia - nuclear expression of Hif1α, magnification 60×. Ir: Irisin.
Figure 3
Figure 3. Comparison of Ir expression by western blotting in the HL-1 cells subjected to hypoxia and normoxia (A). Densitometric analysis of Ir protein levels (B). Ir: Irisin.
Figure 4
Figure 4. Comparison of FNDC5 mRNA expression with use of the real-time PCR method.
See this image and copyright information in PMC

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