Open Access Research

Increased heme-oxygenase 1 expression in mesenchymal stem cell-derived adipocytes decreases differentiation and lipid accumulation via upregulation of the canonical Wnt signaling cascade

Luca Vanella12, Komal Sodhi1, Dong Hyun Kim1, Nitin Puri3, Mani Maheshwari1, Terry D Hinds3, Lars Bellner4, Dov Goldstein5, Stephen J Peterson4, Joseph I Shapiro1 and Nader G Abraham1*

Author Affiliations

1 Joan C. Edwards School of Medicine, Marshall University, 1600 Medical Center Drive, Huntington, WV, 25701-3655, USA

2 Department of Drug Science, Section of Biochemistry, University of Catania, Viale Andrea Doria, 6, 95125 Catania, Italy

3 Department of Physiology and Pharmacology, University of Toledo, 3000 Arlington Avenue, Toledo, OH, 43614-2598, USA

4 Department of Medicine, New York Medical College, Munger Pavilion, Valhalla, NY, 10595, USA

5 Brooklyn Fertility Center, Columbia University, 55 Central Park West, New York, NY 10023, USA

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Stem Cell Research & Therapy 2013, 4:28  doi:10.1186/scrt176

Published: 12 March 2013

Abstract

Introduction

Heme oxygenase (HO), a major cytoprotective enzyme, attenuates oxidative stress and obesity. The canonical Wnt signaling cascade plays a pivotal role in the regulation of adipogenesis. The present study examined the interplay between HO-1and the Wnt canonical pathway in the modulation of adipogenesis in mesenchymal stem cell (MSC)-derived adipocytes.

Methods

To verify the role of HO-1 in generating small healthy adipocytes, cobalt protoporphyrin (CoPP), inducer of HO-1, was used during adipocyte differentiation. Lipid accumulation was measured by Oil red O staining and lipid droplet size was measured by BODIPY staining.

Results

During adipogenesis in vitro, differentiating pre-adipocytes display transient increases in the expression of genes involved in canonical Wnt signaling cascade. Increased levels of HO-1 expression and HO activity resulted in elevated levels of β-catenin, pGSK3β, Wnt10b, Pref-1, and shh along with increased levels of adiponectin (P < 0.05). In addition, induction of HO-1 resulted in a reduction in C/EBPα, PPARγ, Peg-1/Mest, aP2, CD36 expression and lipid accumulation (P < 0.05). Suppression of HO-1 gene by siRNA decreased Wnt10b, pGSK3β and β-catenin expression, and increased lipid accumulation. The canonical Wnt responsive genes, IL-8 and SFRP1, were upregulated by CoPP and their expression was decreased by the concurrent administration of tin mesoporphyrin (SnMP), an inhibitor of HO activity. Furthermore, knockdown of Wnt10b gene expression by using siRNA showed increased lipid accumulation, and this effect was not decreased by concurrent treatment with CoPP. Also our results show that blocking the Wnt 10b antagonist, Dickkopf 1 (Dkk-1), by siRNA decreased lipid accumulation and this effect was further enhanced by concurrent administration of CoPP.

Conclusions

This is the first study to demonstrate that HO-1 acts upstream of canonical Wnt signaling cascade and decreases lipogenesis and adipocyte differentiation suggesting that the HO-1 mediated increase in Wnt10b can modulate the adipocyte phenotype by regulating the transcriptional factors that play a role in adipogenesis. This is evidenced by a decrease in lipid accumulation and inflammatory cytokine levels, increased adiponectin levels and elevation of the expression of genes of the canonical Wnt signaling cascade.