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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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Western blotting results of adipogenesis-specific markers.Representative image of 3 repeats and quantification of (A) PPARγ, (B) C/EBPα, and (C) Acetyl CoA carboxylase adipogenic marker proteins. All values were compared to the adipogenic media control (*p<0.05 and ** p<0.001).
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pone.0118005.g005: Western blotting results of adipogenesis-specific markers.Representative image of 3 repeats and quantification of (A) PPARγ, (B) C/EBPα, and (C) Acetyl CoA carboxylase adipogenic marker proteins. All values were compared to the adipogenic media control (*p<0.05 and ** p<0.001).

Mentions: To study the effect of the Wnt regulators on adipogenesis in ADSCs at the protein level, immunoblotting was performed using antibodies against the key adipogenic marker proteins PPARγ, CCAAT enhancer binding protein (C/EBPα), and acetyl CoA carboxylase. sFRP4 at 1ng/mL concentration significantly upregulated the expression of PPARγ by 1.2-fold, C/EBPα by 1.23-fold, and acetyl CoA carboxylase by 1.3-fold respectively (Fig. 5A, 5B, 5C). The expression of PPARγ, C/EBPα, and acetyl CoA carboxylase was decreased significantly with LiCl (by 1.6, 2.6, 1.9-fold respectively) and BIO (by 7, 17, 5.6-fold respectively) treatments (Fig. 5A, 5B, 5C).


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)

Western blotting results of adipogenesis-specific markers.Representative image of 3 repeats and quantification of (A) PPARγ, (B) C/EBPα, and (C) Acetyl CoA carboxylase adipogenic marker proteins. All values were compared to the adipogenic media control (*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.g005: Western blotting results of adipogenesis-specific markers.Representative image of 3 repeats and quantification of (A) PPARγ, (B) C/EBPα, and (C) Acetyl CoA carboxylase adipogenic marker proteins. All values were compared to the adipogenic media control (*p<0.05 and ** p<0.001).
Mentions: To study the effect of the Wnt regulators on adipogenesis in ADSCs at the protein level, immunoblotting was performed using antibodies against the key adipogenic marker proteins PPARγ, CCAAT enhancer binding protein (C/EBPα), and acetyl CoA carboxylase. sFRP4 at 1ng/mL concentration significantly upregulated the expression of PPARγ by 1.2-fold, C/EBPα by 1.23-fold, and acetyl CoA carboxylase by 1.3-fold respectively (Fig. 5A, 5B, 5C). The expression of PPARγ, C/EBPα, and acetyl CoA carboxylase was decreased significantly with LiCl (by 1.6, 2.6, 1.9-fold respectively) and BIO (by 7, 17, 5.6-fold respectively) treatments (Fig. 5A, 5B, 5C).

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