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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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Related in: MedlinePlus

Oil Red O Staining and quantification on Day 7.(A) Microscopic observations of the stained lipid droplets. Treatment conditions were (i) non-induced control, (ii) adipogenic control, (iii) 10mM LiCl, (iv) 0.5μM BIO, (v) 100pg/mL sFRP4, and (vi) 1ng/mL sFRP4. Scale bar = 250μM (B) 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<0.05 and ** p<0.001).
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pone.0118005.g004: Oil Red O Staining and quantification on Day 7.(A) Microscopic observations of the stained lipid droplets. Treatment conditions were (i) non-induced control, (ii) adipogenic control, (iii) 10mM LiCl, (iv) 0.5μM BIO, (v) 100pg/mL sFRP4, and (vi) 1ng/mL sFRP4. Scale bar = 250μM (B) 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<0.05 and ** p<0.001).

Mentions: Oil Red O staining was performed on Day 7 of adipogenic differentiation, and the stained (red) intracellular lipid droplets were visualised using bright field microscopy. The least lipid droplet accumulation was observed in the groups with LiCl, and BIO and the highest observed in sFRP4 1ng/mL group (Fig. 4A).


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)

Oil Red O Staining and quantification on Day 7.(A) Microscopic observations of the stained lipid droplets. Treatment conditions were (i) non-induced control, (ii) adipogenic control, (iii) 10mM LiCl, (iv) 0.5μM BIO, (v) 100pg/mL sFRP4, and (vi) 1ng/mL sFRP4. Scale bar = 250μM (B) 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<0.05 and ** p<0.001).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0118005.g004: Oil Red O Staining and quantification on Day 7.(A) Microscopic observations of the stained lipid droplets. Treatment conditions were (i) non-induced control, (ii) adipogenic control, (iii) 10mM LiCl, (iv) 0.5μM BIO, (v) 100pg/mL sFRP4, and (vi) 1ng/mL sFRP4. Scale bar = 250μM (B) 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<0.05 and ** p<0.001).
Mentions: Oil Red O staining was performed on Day 7 of adipogenic differentiation, and the stained (red) intracellular lipid droplets were visualised using bright field microscopy. The least lipid droplet accumulation was observed in the groups with LiCl, and BIO and the highest observed in sFRP4 1ng/mL group (Fig. 4A).

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