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Effect of Glucagon-like Peptide-1 on the Differentiation of Adipose-derived Stem Cells into Osteoblasts and Adipocytes.

Lee HM, Joo BS, Lee CH, Kim HY, Ock JH, Lee YS - J Menopausal Med (2015)

Bottom Line: In contrast, GLP-1 significantly suppressed the expression of adipocyte-specific markers, peroxisome proliferator-activated receptor gamma (PPAR-γ), lipoprotein lipase (LPL) and adipocyte protein 2 (AP2).This decreased expression of adipocyte specific markers caused by GLP-1 was significantly reversed by the treatment of extracellular signal-regulated kinase (ERK) inhibitor, PD98059 (P < 0.05).This result demonstrates that GLP-1 stimulates osteoblast differentiation in ADSCs, whereas it inhibits adipocyte differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, Natural Science College, Pusan National University, Busan, Korea.

ABSTRACT

Objectives: Glucagon-like peptide-1 (GLP-1) is an intestinally secreted hormone and it plays an important role in the regulation of glucose homeostasis. However, the possible role of GLP-1 in the differentiation of adipose-derived stem cells (ADSCs) remains unknown. Therefore this study investigated the effect of GLP-1 on the differentiation of ADSCs into osteoblasts and adipocytes.

Methods: ADSCs were isolated from human adipose tissues of the abdomens, cultured and characterized by flow cytometry and multi-lineage potential assay. ADSCs were induced in osteogenic and adipogenic media treated with two different doses (10 and 100 nM) of GLP-1, and then the effect of GLP-1 on differentiation of ADSCs into osteoblast and adipocyte was examined. The signaling pathway involved in these processes was also examined.

Results: Isolated human ADSCs expressed mesenchymal stem cell (MSC) specific markers as well as GLP-1 receptor (GLP-1R) proteins. They also showed multiple-lineage potential of MSC. GLP-1 was upregulated the activity and mRNA expression of osteoblast-specific marker, alkaline phosphatase and the mineralization of calcium. In contrast, GLP-1 significantly suppressed the expression of adipocyte-specific markers, peroxisome proliferator-activated receptor gamma (PPAR-γ), lipoprotein lipase (LPL) and adipocyte protein 2 (AP2). This decreased expression of adipocyte specific markers caused by GLP-1 was significantly reversed by the treatment of extracellular signal-regulated kinase (ERK) inhibitor, PD98059 (P < 0.05).

Conclusion: This result demonstrates that GLP-1 stimulates osteoblast differentiation in ADSCs, whereas it inhibits adipocyte differentiation. The ERK signaling pathway seems to be involved in these differentiation processes mediated by GLP-1.

No MeSH data available.


Effects of glucagon-like peptide-1 (GLP-1) on the osteogenic differentiation of adipose-derived stem cells (ADSCs). The ADSCs were cultured osteogenic induction medium (OIM) treated with various concentration of GLP-1 (0, 10 nM and 100 nM). The media containing GLP-1 was renewed every day. (A) The mRNA level of alkaline phosphatase (ALP) was determined by real time polymerase chain reaction (PCR) and it was normalized by the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). *P < 0.05 (vs. OIM). (B, C) On day 10, ALP activity was measured by ALP staining and enzyme-linked immunosorbent assay (ELISA). ALP was indicated by the red staining. Original magnification, × 100. ALP activity was determined using a colorimetric end point assay measuring the enzyme p-nitrophenol (pNP) in the presence of ALP. (D) On day 14, cells were stained with alizarin red S and calcium deposition nodules were stained as dark red areas. Original magnification, × 100. (E) Bar graph shows quantitative results of (D). †P < 0.05 (vs. OIM).
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Figure 3: Effects of glucagon-like peptide-1 (GLP-1) on the osteogenic differentiation of adipose-derived stem cells (ADSCs). The ADSCs were cultured osteogenic induction medium (OIM) treated with various concentration of GLP-1 (0, 10 nM and 100 nM). The media containing GLP-1 was renewed every day. (A) The mRNA level of alkaline phosphatase (ALP) was determined by real time polymerase chain reaction (PCR) and it was normalized by the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). *P < 0.05 (vs. OIM). (B, C) On day 10, ALP activity was measured by ALP staining and enzyme-linked immunosorbent assay (ELISA). ALP was indicated by the red staining. Original magnification, × 100. ALP activity was determined using a colorimetric end point assay measuring the enzyme p-nitrophenol (pNP) in the presence of ALP. (D) On day 14, cells were stained with alizarin red S and calcium deposition nodules were stained as dark red areas. Original magnification, × 100. (E) Bar graph shows quantitative results of (D). †P < 0.05 (vs. OIM).

Mentions: To examine whether GLP-1 has an effect on osteoblast differentiation, two doses of GLP-1 (10 and 100 nM) were treated to ADSCs cultured in OIM, and then Alizarin Red S staining and ALP staining were assessed after 10 days and 21 days of culture, respectively as the first indicator for osteogenic differentiation. The areas of ALP positive staining (Fig. 3A) and calcified nodules by Alizarin Red S staining (Fig. 3B) were increased in a dose-dependent manner in response to GLP-1, and the significant maximal effect was reached at a concentration of 100 nM GLP-1. Quantitative analysis of calcium deposition showed two-fold and five-fold increase in calcium deposition in 10 nM and 100 nM GLP-1, respectively (Fig. 3C).


Effect of Glucagon-like Peptide-1 on the Differentiation of Adipose-derived Stem Cells into Osteoblasts and Adipocytes.

Lee HM, Joo BS, Lee CH, Kim HY, Ock JH, Lee YS - J Menopausal Med (2015)

Effects of glucagon-like peptide-1 (GLP-1) on the osteogenic differentiation of adipose-derived stem cells (ADSCs). The ADSCs were cultured osteogenic induction medium (OIM) treated with various concentration of GLP-1 (0, 10 nM and 100 nM). The media containing GLP-1 was renewed every day. (A) The mRNA level of alkaline phosphatase (ALP) was determined by real time polymerase chain reaction (PCR) and it was normalized by the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). *P < 0.05 (vs. OIM). (B, C) On day 10, ALP activity was measured by ALP staining and enzyme-linked immunosorbent assay (ELISA). ALP was indicated by the red staining. Original magnification, × 100. ALP activity was determined using a colorimetric end point assay measuring the enzyme p-nitrophenol (pNP) in the presence of ALP. (D) On day 14, cells were stained with alizarin red S and calcium deposition nodules were stained as dark red areas. Original magnification, × 100. (E) Bar graph shows quantitative results of (D). †P < 0.05 (vs. OIM).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 3: Effects of glucagon-like peptide-1 (GLP-1) on the osteogenic differentiation of adipose-derived stem cells (ADSCs). The ADSCs were cultured osteogenic induction medium (OIM) treated with various concentration of GLP-1 (0, 10 nM and 100 nM). The media containing GLP-1 was renewed every day. (A) The mRNA level of alkaline phosphatase (ALP) was determined by real time polymerase chain reaction (PCR) and it was normalized by the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). *P < 0.05 (vs. OIM). (B, C) On day 10, ALP activity was measured by ALP staining and enzyme-linked immunosorbent assay (ELISA). ALP was indicated by the red staining. Original magnification, × 100. ALP activity was determined using a colorimetric end point assay measuring the enzyme p-nitrophenol (pNP) in the presence of ALP. (D) On day 14, cells were stained with alizarin red S and calcium deposition nodules were stained as dark red areas. Original magnification, × 100. (E) Bar graph shows quantitative results of (D). †P < 0.05 (vs. OIM).
Mentions: To examine whether GLP-1 has an effect on osteoblast differentiation, two doses of GLP-1 (10 and 100 nM) were treated to ADSCs cultured in OIM, and then Alizarin Red S staining and ALP staining were assessed after 10 days and 21 days of culture, respectively as the first indicator for osteogenic differentiation. The areas of ALP positive staining (Fig. 3A) and calcified nodules by Alizarin Red S staining (Fig. 3B) were increased in a dose-dependent manner in response to GLP-1, and the significant maximal effect was reached at a concentration of 100 nM GLP-1. Quantitative analysis of calcium deposition showed two-fold and five-fold increase in calcium deposition in 10 nM and 100 nM GLP-1, respectively (Fig. 3C).

Bottom Line: In contrast, GLP-1 significantly suppressed the expression of adipocyte-specific markers, peroxisome proliferator-activated receptor gamma (PPAR-γ), lipoprotein lipase (LPL) and adipocyte protein 2 (AP2).This decreased expression of adipocyte specific markers caused by GLP-1 was significantly reversed by the treatment of extracellular signal-regulated kinase (ERK) inhibitor, PD98059 (P < 0.05).This result demonstrates that GLP-1 stimulates osteoblast differentiation in ADSCs, whereas it inhibits adipocyte differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, Natural Science College, Pusan National University, Busan, Korea.

ABSTRACT

Objectives: Glucagon-like peptide-1 (GLP-1) is an intestinally secreted hormone and it plays an important role in the regulation of glucose homeostasis. However, the possible role of GLP-1 in the differentiation of adipose-derived stem cells (ADSCs) remains unknown. Therefore this study investigated the effect of GLP-1 on the differentiation of ADSCs into osteoblasts and adipocytes.

Methods: ADSCs were isolated from human adipose tissues of the abdomens, cultured and characterized by flow cytometry and multi-lineage potential assay. ADSCs were induced in osteogenic and adipogenic media treated with two different doses (10 and 100 nM) of GLP-1, and then the effect of GLP-1 on differentiation of ADSCs into osteoblast and adipocyte was examined. The signaling pathway involved in these processes was also examined.

Results: Isolated human ADSCs expressed mesenchymal stem cell (MSC) specific markers as well as GLP-1 receptor (GLP-1R) proteins. They also showed multiple-lineage potential of MSC. GLP-1 was upregulated the activity and mRNA expression of osteoblast-specific marker, alkaline phosphatase and the mineralization of calcium. In contrast, GLP-1 significantly suppressed the expression of adipocyte-specific markers, peroxisome proliferator-activated receptor gamma (PPAR-γ), lipoprotein lipase (LPL) and adipocyte protein 2 (AP2). This decreased expression of adipocyte specific markers caused by GLP-1 was significantly reversed by the treatment of extracellular signal-regulated kinase (ERK) inhibitor, PD98059 (P < 0.05).

Conclusion: This result demonstrates that GLP-1 stimulates osteoblast differentiation in ADSCs, whereas it inhibits adipocyte differentiation. The ERK signaling pathway seems to be involved in these differentiation processes mediated by GLP-1.

No MeSH data available.