Korean J Physiol Pharmacol.  2024 May;28(3):285-294. 10.4196/kjpp.2024.28.3.285.

Fibroblast-derived interleukin-6 exacerbates adverse cardiac remodeling after myocardial infarction

• Li H
• Bian Y

• KMID: 2555670
• DOI: http://doi.org/10.4196/kjpp.2024.28.3.285

Abstract

Myocardial infarction is one of the leading causes of mortality globally. Currently, the pleiotropic inflammatory cytokine interleukin-6 (IL-6) is considered to be intimately related to the severity of myocardial injury during myocardial infarction. Interventions targeting IL-6 are a promising therapeutic option for myocardial infarction, but the underlying molecular mechanisms are not well understood. Here, we report the novel role of IL-6 in regulating adverse cardiac remodeling mediated by fibroblasts in a mouse model of myocardial infarction. It was found that the elevated expression of IL-6 in myocardium and cardiac fibroblasts was observed after myocardial infarction. Further, fibroblast-specific knockdown of Il6 significantly attenuated cardiac fibrosis and adverse cardiac remodeling and preserved cardiac function induced by myocardial infarction. Mechanistically, the role of Il6 contributing to cardiac fibrosis depends on signal transduction and activation of transcription (STAT)3 signaling activation. Additionally, Stat3 binds to the Il11 promoter region and contributes to the increased expression of Il11, which exacerbates cardiac fibrosis. In conclusion, these results suggest a novel role for IL-6 derived from fibroblasts in mediating Stat3 activation and substantially augmented Il11 expression in promoting cardiac fibrosis, highlighting its potential as a therapeutic target for cardiac fibrosis.

Keyword

Cardiac fibrosis; Fibroblasts; Interleukin-6; Interleukin-11; Myocardial infarction

Figure

• 

  Fig. 1 Heart injury induces fibroblast interleukin-6 (IL-6) production. (A) Q-PCR analysis of Il6 mRNA expression in myocardial tissues from WT mice after MI or sham operation. (B) ELISA of IL-6 production in myocardial tissue homogenates from WT mice after MI. (C) ELISA of IL-6 production in peripheral blood from mice with myocardial infarction. (D) Q-PCR analysis of Il6 mRNA expression in fibroblasts isolated from WT mice myocardial tissues at day 3 after MI. (E) Q-PCR analysis of Il6 mRNA expression in isolated fibroblasts under hypoxia condition for 1 day. (F) ELISA of IL-6 production in supernatants of isolated fibroblasts under hypoxia condition for 1 day. (G) Q-PCR analysis of Il6 mRNA expression in isolated fibroblasts stimulated with S100A8 for 12 h. (H) ELISA of IL-6 production in supernatants of isolated fibroblasts stimulated with S100A8 for 12 h. n = 5 per group. Data presented as mean ± SD. Unpaired Student’s t-test was performed. WT, wildtype; MI, myocardial infarction; Ctrl, control.

• 

  Fig. 2 Fibroblast-specific interleukin-6 (IL-6) knockout alleviates cardiac dysfunction after MI. (A) Q-PCR analysis of Il6 mRNA expression in fibroblasts isolated from KO or littermate control WT mice. (B) Representative TTC staining images in myocardial tissues from Il6 KO or WT mice at day 14 after MI. (C, D) Echocardiographic measurement of LVEF and LVFS of Il6 KO or WT mice under sham or MI operation for 3 days. (E) Echocardiographic measurement of cardiac output in Il6 KO or WT mice. n = 5 per group. Data presented as mean ± SD. Unpaired Student’s t-test was performed. MI, myocardial infarction; KO, knockout; WT, wildtype; TTC, 2, 3, 5-triphenyl tetrazolium chloride; LVEF, left ventricular ejection fraction; LVFS, left ventricular shortening rate.

• 

  Fig. 3 Fibroblast-specific interleukin-6 (IL-6) knockout alleviates adverse cardiac remodeling after MI. (A) Representative Masson’s Trichrome staining images in myocardial tissues from Il6 KO or WT mice at day 14 after MI (×1.25). (B) Representative immunofluorescence staining of α-SMA in myocardial tissues from Il6 KO or WT mice after MI (×400). (C) Q-PCR analysis of Col1a1, Col3a1, Tgfb and Postn mRNA levels in myocardial tissues from Il6 KO or WT mice after MI. n = 5 per group. Data presented as mean ± SD. Unpaired Student’s t-test was performed. MI, myocardial infarction; KO, knockout; WT, wildtype; α-SMA, α-smooth muscle actin.

• 

  Fig. 4 Fibroblast-derived interleukin-6 (IL-6) promotes cardiac fibrosis via the activation of STAT3. (A) Immunoblot analysis of phosphorylation levels of STAT3 in myocardial tissues from WT mice after MI. (B) Immunoblot analysis of Collagen I and Collagen III in myocardial tissues from WT mice after MI. (C) Immunoblot analysis of phosphorylation levels of STAT3 in myocardial tissues from Ii6 KO or WT mice after MI. (D) Immunoblot analysis of Collagen I and Collagen III in myocardial tissues from Ii6 KO or WT mice after MI. (E) Immunoblot analysis of phosphorylation levels of STAT3 in myocardial tissues from Ctrl or STAT3-IN-3 treated mice after MI. (F) Immunoblot analysis of Collagen I and Collagen III in myocardial tissues from Ctrl or STAT3-IN-3 treated mice after MI. Similar results were obtained from three independent experiments. Data presented as mean ± SD. Unpaired Student’s t-test was performed. STAT3, signal transduction and activation of transcription 3; WT, wildtype; MI, myocardial infarction; KO, knockout; Ctrl, control. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

• 

  Fig. 5 STAT3 promotes the expression of Il11 in fibroblasts. (A) IGV analysis of STAT3 (GSE211111) signals in the gene locus of Il11. (B) ChIP analysis of STAT3 enrichment level at the Il11promoter in isolated fibroblasts after MI (n = 4 per group). (C) Q-PCR analysis of Il11 mRNA expression in myocardial tissues from wild-type mice after MI or sham operation (n = 5 per group). (D) Q-PCR analysis of Il11 mRNA expression in fibroblasts treated with IL-6 or control PBS for 12 h (n = 5 per group). (E) Q-PCR analysis of Il11 mRNA expression in myocardial tissues from KO or WT mice after MI or sham operation (n = 5 per group). (F) Q-PCR analysis of Il11 mRNA expression in myocardial tissues from Ctrl or STAT3-IN-3 treated mice after MI (n = 5 per group). (G) Q-PCR analysis of Col1a1, Col3a1, Tgfb and Postn mRNA levels in fibroblast transfection with Il11 siRNA or control siRNA followed by treatment with IL-6 for 12 h (n = 5 per group). Data presented as mean ± SD. Unpaired Student’s t-test was performed. STAT3, signal transduction and activation of transcription 3; IGV, integrative genomics viewer; ChIP, chromatin immunoprecipitation; MI, myocardial infarction; IL-6, interleukin-6; KO, knockout; WT, wildtype; Ctrl, control.

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