Int J Stem Cells.  2021 Nov;14(4):455-464. 10.15283/ijsc21005.

Irisin Enhances Angiogenesis of Mesenchymal Stem Cells to Promote Cardiac Function in Myocardial Infarction via PI3k/Akt Activation

• Yang F
• Wang Z
• Li B
• He Y
• Du F
• Tian S
• Zhang Y
• Yang Y

• KMID: 2522563
• DOI: http://doi.org/10.15283/ijsc21005

Abstract

Background and Objectives
With the growing incidence of acute myocardial infarction (MI), angiogenesis is vital for cardiac function post-MI. The role of bone marrow mesenchymal stem cells (BMSCs) in angiogenesis has been previously confirmed. Irisin is considered a potential vector for angiogenesis. The objective of the present study was to investigate the potential role of irisin in the angiogenesis of BMSCs.
Methods and Results
In vivo, irisin-treated BMSCs (BMSCs＋irisin) were transplanted into an MI mouse model. On day 28 post-MI, blood vessel markers were detected, and cardiac function and infarct areas of mice were evaluated. In vitro, paracrine effects were assessed by examining tube formation in human umbilical vein endothelial cells (HUVECs) co-cultured with the BMSCs＋irisin supernatant. The scratch wound-healing assay was performed to evaluate HUVEC migration. Western blotting was performed to determine PI3k/Akt pathway activation in the BMSCs＋irisin group. Transplantation of BMSCs＋irisin promoted greater angiogenesis, resulting in better cardiac function in the MI mouse model than in controls. In the BMSC＋irisin group, HUVECs demonstrated enhanced tube formation and migration. Activation of the PI3k/Akt pathway was found to be involved in mediating the role of irisin in the angiogenesis of BMSCs.
Conclusions
In cardiovascular diseases such as MI, irisin administration can enhance angiogenesis of BMSCs and pro-mote cardiac function via the PI3k/Akt pathway, optimizing the therapeutic effect based on BMSCs transplantation.

Keyword

Irisin; Angiogenesis; Mesenchymal stem cells; PI3k/Akt pathway

Figure

• 

  Fig. 1 Irisin-treated BMSCs rescued the cardiac function in MI mouse in vivo. (a) Representative images of echocardiography showing the improved cardiac function in BMSCs treated with irisin. (b) The ejection fraction (EF) and fractional shortening (FS) are gradually recovered in the BMSCs＋irisin group when compared with other groups (n=8 for Sham, 5 for DMEM, 6 for BMSCs＋irisin and BMSCs＋Con). (c) The infarct area was examined by Sirius Red staining 28 days post-MI (n=6 for every group). (d) The infarct size in hearts was quantified using Image-Pro. All data were measured as mean±standard error of the mean (SEM). *p<0.05. BMSCs, bone marrow mesenchymal stem cells; MI, myocardial infarction; DMEM, Dulbecco’s Modified Eagle Medium.

• 

  Fig. 2 Irisin-treated BMSCs enhance angiogenesis. (a∼c) Representative images of vascular markers including α-SMA, vWF and CD31 were captured in the border zone of irisin-treated BMSCs, control, and DMEM groups. (d) The vascular density was measured in at least 5 high-power fields per section (n=8 for DMEM, 7 for BMSCs＋irisin and BMSCs＋Con). Scale bar, 100 μm. Data are presented as mean±standard error of the mean (SEM). *p<0.05. BMSCs, bone marrow mesenchymal stem cells; DMEM, Dulbecco’s Modified Eagle Medium; α-SMA, alpha-smooth muscle actin; vWF, von Willebrand factor.

• 

  Fig. 3 Irisin promotes the paracrine efficacy of BMSCs through the PI3K/AKT pathway. (a) Conditioned media of equivalent cell numbers (1×106 cells) were collected for irisin-treated BMSCs and control groups. By employing HUVECs, tube forma-tion was assessed using the supernatant of BMSCs treated with irisin and control, irisin alone, and control alone. Scale bar, 50 μm. (b) The wound-healing assay was performed to demonstrate HUVEC migration using the above conditioned media, respectively. Scale bar, 243.2 μm. (c) The number of branch points was quantified by Image-Pro software and is shown in bar graphs (n=3 separate studies). (d) Quantification of HUVEC migration by Image-Pro software (n=3). (e) Western blot analyses of pPI3K, PI3K, p-AKT, AKT, p-ERK1/2, ERK1/2, p-P38, P38, p-JNK, and JNK pathway were detected in irisin-treated BMSCs and control groups. All data were measured as mean±standard error of the mean (SEM). *p<0.05. BMSCs, bone marrow mesenchymal stem cells; HUVECs, human umbilical vein endothelial cells.

• 

  Fig. 4 The PI3K/AKT pathway is required to mediate the paracrine efficacy of BMSCs. (a) Representative images of tube formation were analyzed using HUVECs co-cultured with the supernatant of BMSCs treated with irisin, LY294002, irisin＋LY294002, or control, respectively. Scale bar, 50 μm. (b) The number of branch points in the above groups was quantified by Image-Pro software and is shown in bar graphs (n=3). (c) The wound-healing assay assessed HUVEC migration using the above conditioned media, respec-tively. Scale bar, 243.2 μm. (d) Quantification of HUVEC migration (n=3) by Image-Pro software. Data are presented as mean±standard error of the mean (SEM). *p<0.05. BMSCs, bone marrow mesenchymal stem cells; HUVECs, human umbilical vein endothelial cells.

Reference

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