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
. 2024 Oct;15(5):404-414.
doi: 10.14740/cr1705. Epub 2024 Oct 11.

Fibroblast Growth Factor 21 Suppressed Neutrophil Extracellular Traps Induced by Myocardial Ischemia/Reperfusion Injury via Adenosine Monophosphate-Activated Protein Kinase

Ling Yun Gu  1 , Cheng Gao Jia  2 , Zuo Zhen Sheng  1 , Wen Long Jiang  1 , Zhuo Wen Xu  2 , Wei Zhang Li  2 , Jun You Cui  1 , Hua Zhang  1   2
Affiliations

Fibroblast Growth Factor 21 Suppressed Neutrophil Extracellular Traps Induced by Myocardial Ischemia/Reperfusion Injury via Adenosine Monophosphate-Activated Protein Kinase

Ling Yun Gu et al. Cardiol Res. 2024 Oct.

Abstract

Background: Previous investigations have established the anti-inflammatory properties of fibroblast growth factor 21 (FGF21). However, the specific mechanism through which FGF21 mitigates myocardial ischemia/reperfusion (I/R) injury by inhibiting neutrophil extracellular traps (NETs) remains unclear.

Methods: A mice model of myocardial I/R injury was induced, and myocardial tissue was stained with immunofluorescence to assess NETs. Serum NETs levels were quantified using a PicoGreen kit. In addition, the expression levels of adenosine monophosphate (AMP)-activated protein kinase (AMPK) and FGF21 were evaluated by Wes fully automated protein blotting quantitative analysis system. Moreover, a hypoxia/reoxygenation (H/R) model was established using AMPK inhibitor and agonist pretreated H9c2 cells to further explore the relationship between FGF21 and AMPK.

Results: Compared with the control group, serum NETs levels were significantly higher in I/R mice, and a large number of NETs were formed in myocardial tissues (97.63 ± 11.45 vs. 69.65 ± 3.33, P < 0.05). However, NETs levels were reversed in FGF21 pretreated mice (P < 0.05). Further studies showed that FGF21 enhanced AMPK expression, which was significantly increased after inhibition of AMPK and decreased after promotion of AMPK (P < 0.05).

Conclusions: FGF21 may exert cardioprotective effects by inhibiting I/R injury-induced NETs via AMPK.

Keywords: AMP-activated protein kinase; Fibroblast growth factor 21; Myocardial ischemia/reperfusion injury; Neutrophil extracellular traps.

PubMed Disclaimer

Conflict of interest statement

The authors declare that they have no conflict of interest.

Figures

Figure 1
Figure 1
FGF21 inhibited I/R injury in myocardial tissue (TTC staining). (a) TTC staining of myocardial tissue. (b) Myocardial infarction volume calculated based on TTC staining. *P < 0.05 versus control group; #P < 0.05 versus control + FGF21 group; &P < 0.05 versus I/R group. FGF21: fibroblast growth factor 21; I/R: ischemia/reperfusion; TTC: 2,3,5-triphenyl tetrazolium chloride.
Figure 2
Figure 2
Myocardial tissue after myocardial I/R injury in mice pretreated with FGF21 (H&E staining, × 400). The black arrows represent obvious pathological changes in myocardial tissue. FGF21: fibroblast growth factor 21; H&E: hematoxylin and eosin; I/R: ischemia/reperfusion.
Figure 3
Figure 3
NETs formation in myocardial necrotic tissues was detected after the establishment of a myocardial I/R injury model in FGF21 pretreated mice (immunofluorescence, × 400). *P < 0.05 versus control group; #P < 0.05 versus control + FGF21 group; &P < 0.05 versus I/R group. FGF21: fibroblast growth factor 21; I/R: ischemia/reperfusion; NETs: neutrophil extracellular traps.
Figure 4
Figure 4
Measurement of peripheral serum dsDNA levels in FGF21 pretreated mice using the PicoGreen assay. *P < 0.05 versus control group; #P < 0.05 versus control + FGF21 group; &P < 0.05 versus I/R group. FGF21: fibroblast growth factor 21; I/R: ischemia/reperfusion.
Figure 5
Figure 5
Myocardial tissue after myocardial I/R injury in mice pretreated with AMPK inhibitor (H&E staining, × 400). The black arrows represent obvious pathological changes in myocardial tissue. AMPK: AMP-activated protein kinase; H&E: hematoxylin and eosin; I/R: ischemia/reperfusion.
Figure 6
Figure 6
NETs formation in myocardial necrotic tissues was detected after the establishment of a myocardial I/R injury model in compound C pretreated mice (immunofluorescence, × 400). *P < 0.05 versus control group; #P < 0.05 versus control + FGF21 group; &P < 0.05 versus I/R group. FGF21: fibroblast growth factor 21; I/R: ischemia/reperfusion; NETs: neutrophil extracellular traps.
Figure 7
Figure 7
Measurement of peripheral serum dsDNA levels in compound C pretreated mice using the PicoGreen assay. *P < 0.05 versus control group; #P < 0.05 versus control + FGF21 group; &P < 0.05 versus I/R group. FGF21: fibroblast growth factor 21; I/R: ischemia/reperfusion.
Figure 8
Figure 8
The interaction of FGF21 and AMPK was detected in a mouse model of myocardial I/R injury or an H/R model of H9c2 cells. (a) AMPK protein expression levels were measured in mice preinjected with FGF21. (b) FGF21 protein expression levels were measured in mice preinjected with AMPK inhibitor compound C. (c) FGF21 protein expression levels were measured in H9c2 cells pretreated with compound C, an inhibitor of AMPK, or with AICAR, an agonist. *P < 0.05 versus control; #P < 0.05 versus I/R group; &P < 0.05 versus H/R + compound C group. AMPK: AMP-activated protein kinase; FGF21: fibroblast growth factor 21; H/R: hypoxia/reoxygenation; I/R: ischemia/reperfusion.
Figure 9
Figure 9
AMPK inhibited H/R injury in H9c2 cells in a time-dependent manner. *P < 0.05 versus control; #P < 0.05 versus H/R group; &P < 0.05 versus H/R + compound C group. AMPK: AMP-activated protein kinase; H/R: hypoxia/reoxygenation.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Bhatt DL, Lopes RD, Harrington RA. Diagnosis and treatment of acute coronary syndromes: a review. JAMA. 2022;327(7):662–675. doi: 10.1001/jama.2022.0358. - DOI - PubMed
    1. Ibanez B, Heusch G, Ovize M, Van de Werf F. Evolving therapies for myocardial ischemia/reperfusion injury. J Am Coll Cardiol. 2015;65(14):1454–1471. doi: 10.1016/j.jacc.2015.02.032. - DOI - PubMed
    1. Frank A, Bonney M, Bonney S, Weitzel L, Koeppen M, Eckle T. Myocardial ischemia reperfusion injury: from basic science to clinical bedside. Semin Cardiothorac Vasc Anesth. 2012;16(3):123–132. doi: 10.1177/1089253211436350. - DOI - PMC - PubMed
    1. Frohlich GM, Meier P, White SK, Yellon DM, Hausenloy DJ. Myocardial reperfusion injury: looking beyond primary PCI. Eur Heart J. 2013;34(23):1714–1722. doi: 10.1093/eurheartj/eht090. - DOI - PubMed
    1. Hausenloy DJ, Yellon DM. Myocardial ischemia-reperfusion injury: a neglected therapeutic _target. J Clin Invest. 2013;123(1):92–100. doi: 10.1172/JCI62874. - DOI - PMC - PubMed

Grants and funding

This research was supported by the development fund of Affiliated Hospital of Xuzhou Medical University (grant no. XYFY2021029), Medical Research Program of Jiangsu Provincial Health and Wellness Commission (grant no. Z2021042), and Jiangyin Young and Middle-aged Reserve Excellent Talents Program (grant no. JYROYT202309).

LinkOut - more resources

  NODES
twitter 2