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Irisin Induces Angiogenesis in Human Umbilical Vein Endothelial Cells In Vitro and in Zebrafish Embryos In Vivo via Activation of the ERK Signaling Pathway.

Wu F, Song H, Zhang Y, Zhang Y, Mu Q, Jiang M, Wang F, Zhang W, Li L, Li H, Wang Y, Zhang M, Li S, Yang L, Meng Y, Tang D - PLoS ONE (2015)

Bottom Line: Although our recent evidence supported the contribution of irisin to vascular endothelial cell (ECs) proliferation and apoptosis, further research of irisin involvement in the angiogenesis of ECs was not conclusive.We also found that irisin activated extracellular signal-related kinase (ERK) signaling pathways.Inhibition of ERK signaling by using U0126 decreased the pro-migration and pro-angiogenic effect of irisin on HUVEC.

View Article: PubMed Central - PubMed

Affiliation: Center for Stem Cell & Regenerative Medicine, The Second Hospital of Shandong University, Jinan, 250012, P.R. China.

ABSTRACT
As a link between exercise and metabolism, irisin is assumed to be involved in increased total body energy expenditure, reduced body weight, and increased insulin sensitivity. Although our recent evidence supported the contribution of irisin to vascular endothelial cell (ECs) proliferation and apoptosis, further research of irisin involvement in the angiogenesis of ECs was not conclusive. In the current study, it was found that irisin promoted Human Umbilical Vein Endothelial Cell (HUVEC) angiogenesis via increasing migration and tube formation, and attenuated chemically-induced intersegmental vessel (ISV) angiogenic impairment in transgenic TG (fli1: GFP) zebrafish. It was further demonstrated that expression of matrix metalloproteinase (MMP) 2 and 9 were also up-regulated in endothelial cells. We also found that irisin activated extracellular signal-related kinase (ERK) signaling pathways. Inhibition of ERK signaling by using U0126 decreased the pro-migration and pro-angiogenic effect of irisin on HUVEC. Also, U0126 inhibited the elevated expression of MMP-2 and MMP-9 when they were treated with irisin. In summary, these findings provided direct evidence that irisin may play a pivotal role in maintaining endothelium homeostasis by promoting endothelial cell angiogenesis via the ERK signaling pathway.

No MeSH data available.


Related in: MedlinePlus

Effect of irisin on matrix metalloproteinases (MMPs) expression and gelatinolytic activity (A) qPCR analysis of MMPs, TIMP1, and endostatin mRNA expression after treatment with or without irisin (10 and 20 nM) for 24 h (n = 3).(B) qPCR analysis of MMPs expression in zebrafish embryos after treatment as described in methods and materials (n = 3). (C) Expression of MMP-2 and MMP-9 were determined by Western blot and normalized with the β-actin level. (D) and (F) Protein bands were quantified by densitometric analysis. (E) MMP-2 and MMP-9 activities were analyzed by gelatin zymography. Data are presented as the mean± SE of triplicates. ** P< 0.01 vs. control group, determined by unpaired two-tailed Student’s t-test.
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pone.0134662.g004: Effect of irisin on matrix metalloproteinases (MMPs) expression and gelatinolytic activity (A) qPCR analysis of MMPs, TIMP1, and endostatin mRNA expression after treatment with or without irisin (10 and 20 nM) for 24 h (n = 3).(B) qPCR analysis of MMPs expression in zebrafish embryos after treatment as described in methods and materials (n = 3). (C) Expression of MMP-2 and MMP-9 were determined by Western blot and normalized with the β-actin level. (D) and (F) Protein bands were quantified by densitometric analysis. (E) MMP-2 and MMP-9 activities were analyzed by gelatin zymography. Data are presented as the mean± SE of triplicates. ** P< 0.01 vs. control group, determined by unpaired two-tailed Student’s t-test.

Mentions: Mounting evidence supports the view that matrix metalloproteinases (MMPs) mediate the process of remodeling the extracellular matrix (ECM) and is a prerequisite for angiogenesis [19, 23]. To determine which MMPs can be influenced by irisin in HUVEC, we examined mRNA expression of MMPs and other genes which may play a critical role in the progress of cells migration. The quantitative real-time PCR results showed that irisin significantly stimulated expression of MMP-2 and MMP-9 and slightly reduced expression of TIMP-1 in HUVEC (Fig 4A). TIMPs play a role in regulating MMP activation in several different tissues and suppress the activity of MMPs [24]. The q-PCR results of MMPs expression in zebrafish embryos were also in line with what we found in HUVEC (Fig 3B). To verify the real-time PCR results, Western blot showed that MMP-2 and MMP-9 protein levels were also highly up-regulated after treatment with irisin for 24 h (Fig 4C and 4D).Gelatin-zymography analysis showed that irisin potently increased the gelatinolytic activity of MMP-2 and MMP-9 secreted from HUVEC (Fig 4E and 4F).


Irisin Induces Angiogenesis in Human Umbilical Vein Endothelial Cells In Vitro and in Zebrafish Embryos In Vivo via Activation of the ERK Signaling Pathway.

Wu F, Song H, Zhang Y, Zhang Y, Mu Q, Jiang M, Wang F, Zhang W, Li L, Li H, Wang Y, Zhang M, Li S, Yang L, Meng Y, Tang D - PLoS ONE (2015)

Effect of irisin on matrix metalloproteinases (MMPs) expression and gelatinolytic activity (A) qPCR analysis of MMPs, TIMP1, and endostatin mRNA expression after treatment with or without irisin (10 and 20 nM) for 24 h (n = 3).(B) qPCR analysis of MMPs expression in zebrafish embryos after treatment as described in methods and materials (n = 3). (C) Expression of MMP-2 and MMP-9 were determined by Western blot and normalized with the β-actin level. (D) and (F) Protein bands were quantified by densitometric analysis. (E) MMP-2 and MMP-9 activities were analyzed by gelatin zymography. Data are presented as the mean± SE of triplicates. ** P< 0.01 vs. control group, determined by unpaired two-tailed Student’s t-test.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4524626&req=5

pone.0134662.g004: Effect of irisin on matrix metalloproteinases (MMPs) expression and gelatinolytic activity (A) qPCR analysis of MMPs, TIMP1, and endostatin mRNA expression after treatment with or without irisin (10 and 20 nM) for 24 h (n = 3).(B) qPCR analysis of MMPs expression in zebrafish embryos after treatment as described in methods and materials (n = 3). (C) Expression of MMP-2 and MMP-9 were determined by Western blot and normalized with the β-actin level. (D) and (F) Protein bands were quantified by densitometric analysis. (E) MMP-2 and MMP-9 activities were analyzed by gelatin zymography. Data are presented as the mean± SE of triplicates. ** P< 0.01 vs. control group, determined by unpaired two-tailed Student’s t-test.
Mentions: Mounting evidence supports the view that matrix metalloproteinases (MMPs) mediate the process of remodeling the extracellular matrix (ECM) and is a prerequisite for angiogenesis [19, 23]. To determine which MMPs can be influenced by irisin in HUVEC, we examined mRNA expression of MMPs and other genes which may play a critical role in the progress of cells migration. The quantitative real-time PCR results showed that irisin significantly stimulated expression of MMP-2 and MMP-9 and slightly reduced expression of TIMP-1 in HUVEC (Fig 4A). TIMPs play a role in regulating MMP activation in several different tissues and suppress the activity of MMPs [24]. The q-PCR results of MMPs expression in zebrafish embryos were also in line with what we found in HUVEC (Fig 3B). To verify the real-time PCR results, Western blot showed that MMP-2 and MMP-9 protein levels were also highly up-regulated after treatment with irisin for 24 h (Fig 4C and 4D).Gelatin-zymography analysis showed that irisin potently increased the gelatinolytic activity of MMP-2 and MMP-9 secreted from HUVEC (Fig 4E and 4F).

Bottom Line: Although our recent evidence supported the contribution of irisin to vascular endothelial cell (ECs) proliferation and apoptosis, further research of irisin involvement in the angiogenesis of ECs was not conclusive.We also found that irisin activated extracellular signal-related kinase (ERK) signaling pathways.Inhibition of ERK signaling by using U0126 decreased the pro-migration and pro-angiogenic effect of irisin on HUVEC.

View Article: PubMed Central - PubMed

Affiliation: Center for Stem Cell & Regenerative Medicine, The Second Hospital of Shandong University, Jinan, 250012, P.R. China.

ABSTRACT
As a link between exercise and metabolism, irisin is assumed to be involved in increased total body energy expenditure, reduced body weight, and increased insulin sensitivity. Although our recent evidence supported the contribution of irisin to vascular endothelial cell (ECs) proliferation and apoptosis, further research of irisin involvement in the angiogenesis of ECs was not conclusive. In the current study, it was found that irisin promoted Human Umbilical Vein Endothelial Cell (HUVEC) angiogenesis via increasing migration and tube formation, and attenuated chemically-induced intersegmental vessel (ISV) angiogenic impairment in transgenic TG (fli1: GFP) zebrafish. It was further demonstrated that expression of matrix metalloproteinase (MMP) 2 and 9 were also up-regulated in endothelial cells. We also found that irisin activated extracellular signal-related kinase (ERK) signaling pathways. Inhibition of ERK signaling by using U0126 decreased the pro-migration and pro-angiogenic effect of irisin on HUVEC. Also, U0126 inhibited the elevated expression of MMP-2 and MMP-9 when they were treated with irisin. In summary, these findings provided direct evidence that irisin may play a pivotal role in maintaining endothelium homeostasis by promoting endothelial cell angiogenesis via the ERK signaling pathway.

No MeSH data available.


Related in: MedlinePlus