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Wnt antagonist secreted frizzled-related protein 4 upregulates adipogenic differentiation in human adipose tissue-derived mesenchymal stem cells.

Visweswaran M, Schiefer L, Arfuso F, Dilley RJ, Newsholme P, Dharmarajan A - PLoS ONE (2015)

Bottom Line: We also examined the effect of Wnt inhibition using secreted frizzled-related protein 4 (sFRP4), which we have previously shown to be pro-apoptotic, anti-angiogenic, and anti-tumorigenic.Wnt stimulation in LiCl and BIO-treated ADSCs resulted in a significant reduction (2.7-fold and 12-fold respectively) in lipid accumulation as measured by Oil red O staining while Wnt inhibition with sFRP4 induced a 1.5-fold increase in lipid accumulation.In contrast, the expression of adipogenic proteins (PPARγ, C/EBPα, and acetyl CoA carboxylase) were decreased significantly with LiCl (by 1.6, 2.6, and 1.9-fold respectively) and BIO (by 7, 17, and 5.6-fold respectively) treatments.

View Article: PubMed Central - PubMed

Affiliation: School of Biomedical Sciences, Curtin Health Innovation Research Institute, Curtin University, Perth, Western Australia, Australia.

ABSTRACT
With more than 1.4 billion overweight or obese adults worldwide, obesity and progression of the metabolic syndrome are major health and economic challenges. To address mechanisms of obesity, adipose tissue-derived mesenchymal stem cells (ADSCs) are being studied to detail the molecular mechanisms involved in adipogenic differentiation. Activation of the Wnt signalling pathway has inhibited adipogenesis from precursor cells. In our study, we examined this anti-adipogenic effect in further detail stimulating Wnt with lithium chloride (LiCl) and 6-bromo indirubin 3'oxime (BIO). We also examined the effect of Wnt inhibition using secreted frizzled-related protein 4 (sFRP4), which we have previously shown to be pro-apoptotic, anti-angiogenic, and anti-tumorigenic. Wnt stimulation in LiCl and BIO-treated ADSCs resulted in a significant reduction (2.7-fold and 12-fold respectively) in lipid accumulation as measured by Oil red O staining while Wnt inhibition with sFRP4 induced a 1.5-fold increase in lipid accumulation. Furthermore, there was significant 1.2-fold increase in peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT/enhancer binding protein alpha (C/EBPα), and 1.3-fold increase in acetyl CoA carboxylase protein levels. In contrast, the expression of adipogenic proteins (PPARγ, C/EBPα, and acetyl CoA carboxylase) were decreased significantly with LiCl (by 1.6, 2.6, and 1.9-fold respectively) and BIO (by 7, 17, and 5.6-fold respectively) treatments. These investigations demonstrate interplay between Wnt antagonism and Wnt activation during adipogenesis and indicate pathways for therapeutic intervention to control this process.

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Morphology of ADSCs during adipogenic differentiation at Day 7.(A) Before and (B) after Oil red O staining. Treatment conditions were (i) non-induced control media, (ii) adipogenic control, (iii) 0.5μM BIO, (iv) 1ng/mL sFRP4, and (v) 0.5μM BIO + 1ng/mL sFRP4 in both (A) and (B). Scale bar = 10μM in (A) and 250μM in (B). (C) Quantification of the stained lipid droplets were performed using the eluted Oil red O stain via measuring absorbance at 510nm. The readings were normalised to background values of non-induced control ADSCs. The values of all treatment conditions were compared to the adipogenic control group (** p<0.001).
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pone.0118005.g006: Morphology of ADSCs during adipogenic differentiation at Day 7.(A) Before and (B) after Oil red O staining. Treatment conditions were (i) non-induced control media, (ii) adipogenic control, (iii) 0.5μM BIO, (iv) 1ng/mL sFRP4, and (v) 0.5μM BIO + 1ng/mL sFRP4 in both (A) and (B). Scale bar = 10μM in (A) and 250μM in (B). (C) Quantification of the stained lipid droplets were performed using the eluted Oil red O stain via measuring absorbance at 510nm. The readings were normalised to background values of non-induced control ADSCs. The values of all treatment conditions were compared to the adipogenic control group (** p<0.001).

Mentions: To study the simultaneous effect of Wnt activator and antagonist on adipogenic differentiation, the degree of lipid accumulation in the presence of both BIO at 0.5μM and sFRP4 at 1ng/mL (B+S) was observed. The morphology of the cells changed from the spindle-like shape to a rounded shape on Day 1 of differentiation in both BIO-only and B+S treatment groups. However, on Day 7 the lipid accumulation remained unchanged in the B+S treatment group when compared to the BIO-only group (Fig. 6A). This was confirmed by Oil red O staining and its quantification (Fig. 6B and 6C), which showed no significant difference in lipid accumulation between BIO-only and B+S treatment groups. This indicated the strong inhibitory action of BIO on adipogenesis which was not reversed by the adipogenic-promoting effect of sFRP4.


Wnt antagonist secreted frizzled-related protein 4 upregulates adipogenic differentiation in human adipose tissue-derived mesenchymal stem cells.

Visweswaran M, Schiefer L, Arfuso F, Dilley RJ, Newsholme P, Dharmarajan A - PLoS ONE (2015)

Morphology of ADSCs during adipogenic differentiation at Day 7.(A) Before and (B) after Oil red O staining. Treatment conditions were (i) non-induced control media, (ii) adipogenic control, (iii) 0.5μM BIO, (iv) 1ng/mL sFRP4, and (v) 0.5μM BIO + 1ng/mL sFRP4 in both (A) and (B). Scale bar = 10μM in (A) and 250μM in (B). (C) Quantification of the stained lipid droplets were performed using the eluted Oil red O stain via measuring absorbance at 510nm. The readings were normalised to background values of non-induced control ADSCs. The values of all treatment conditions were compared to the adipogenic control group (** p<0.001).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4340908&req=5

pone.0118005.g006: Morphology of ADSCs during adipogenic differentiation at Day 7.(A) Before and (B) after Oil red O staining. Treatment conditions were (i) non-induced control media, (ii) adipogenic control, (iii) 0.5μM BIO, (iv) 1ng/mL sFRP4, and (v) 0.5μM BIO + 1ng/mL sFRP4 in both (A) and (B). Scale bar = 10μM in (A) and 250μM in (B). (C) Quantification of the stained lipid droplets were performed using the eluted Oil red O stain via measuring absorbance at 510nm. The readings were normalised to background values of non-induced control ADSCs. The values of all treatment conditions were compared to the adipogenic control group (** p<0.001).
Mentions: To study the simultaneous effect of Wnt activator and antagonist on adipogenic differentiation, the degree of lipid accumulation in the presence of both BIO at 0.5μM and sFRP4 at 1ng/mL (B+S) was observed. The morphology of the cells changed from the spindle-like shape to a rounded shape on Day 1 of differentiation in both BIO-only and B+S treatment groups. However, on Day 7 the lipid accumulation remained unchanged in the B+S treatment group when compared to the BIO-only group (Fig. 6A). This was confirmed by Oil red O staining and its quantification (Fig. 6B and 6C), which showed no significant difference in lipid accumulation between BIO-only and B+S treatment groups. This indicated the strong inhibitory action of BIO on adipogenesis which was not reversed by the adipogenic-promoting effect of sFRP4.

Bottom Line: We also examined the effect of Wnt inhibition using secreted frizzled-related protein 4 (sFRP4), which we have previously shown to be pro-apoptotic, anti-angiogenic, and anti-tumorigenic.Wnt stimulation in LiCl and BIO-treated ADSCs resulted in a significant reduction (2.7-fold and 12-fold respectively) in lipid accumulation as measured by Oil red O staining while Wnt inhibition with sFRP4 induced a 1.5-fold increase in lipid accumulation.In contrast, the expression of adipogenic proteins (PPARγ, C/EBPα, and acetyl CoA carboxylase) were decreased significantly with LiCl (by 1.6, 2.6, and 1.9-fold respectively) and BIO (by 7, 17, and 5.6-fold respectively) treatments.

View Article: PubMed Central - PubMed

Affiliation: School of Biomedical Sciences, Curtin Health Innovation Research Institute, Curtin University, Perth, Western Australia, Australia.

ABSTRACT
With more than 1.4 billion overweight or obese adults worldwide, obesity and progression of the metabolic syndrome are major health and economic challenges. To address mechanisms of obesity, adipose tissue-derived mesenchymal stem cells (ADSCs) are being studied to detail the molecular mechanisms involved in adipogenic differentiation. Activation of the Wnt signalling pathway has inhibited adipogenesis from precursor cells. In our study, we examined this anti-adipogenic effect in further detail stimulating Wnt with lithium chloride (LiCl) and 6-bromo indirubin 3'oxime (BIO). We also examined the effect of Wnt inhibition using secreted frizzled-related protein 4 (sFRP4), which we have previously shown to be pro-apoptotic, anti-angiogenic, and anti-tumorigenic. Wnt stimulation in LiCl and BIO-treated ADSCs resulted in a significant reduction (2.7-fold and 12-fold respectively) in lipid accumulation as measured by Oil red O staining while Wnt inhibition with sFRP4 induced a 1.5-fold increase in lipid accumulation. Furthermore, there was significant 1.2-fold increase in peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT/enhancer binding protein alpha (C/EBPα), and 1.3-fold increase in acetyl CoA carboxylase protein levels. In contrast, the expression of adipogenic proteins (PPARγ, C/EBPα, and acetyl CoA carboxylase) were decreased significantly with LiCl (by 1.6, 2.6, and 1.9-fold respectively) and BIO (by 7, 17, and 5.6-fold respectively) treatments. These investigations demonstrate interplay between Wnt antagonism and Wnt activation during adipogenesis and indicate pathways for therapeutic intervention to control this process.

Show MeSH
Related in: MedlinePlus