Reciprocal Interferences of TNF-α and Wnt1/β-Catenin Signaling Axes Shift Bone Marrow-Derived Stem Cells Towards Osteoblast Lineage after Ethanol Exposure
Background/Aims: We have reported in a separate study that alcohol exposure triggers activation of the TNF-α signaling pathway leading to an adverse shift of multipotential mesenchymal stem cells in bone marrow (BMSCs) away from osteogenesis towards adipogenesis. However, inhibition of TNF-α signali...
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| Format: | Article |
| Language: | English |
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Cell Physiol Biochem Press GmbH & Co KG
2013-09-01
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| Series: | Cellular Physiology and Biochemistry |
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| Online Access: | http://www.karger.com/Article/FullText/354477 |
| _version_ | 1811214336674234368 |
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| author | Yueping Chen Liang Chen Qingshui Yin Hui Gao Panfeng Dong Xiaoyun Zhang Jie Kang |
| author_facet | Yueping Chen Liang Chen Qingshui Yin Hui Gao Panfeng Dong Xiaoyun Zhang Jie Kang |
| author_sort | Yueping Chen |
| collection | DOAJ |
| description | Background/Aims: We have reported in a separate study that alcohol exposure triggers activation of the TNF-α signaling pathway leading to an adverse shift of multipotential mesenchymal stem cells in bone marrow (BMSCs) away from osteogenesis towards adipogenesis. However, inhibition of TNF-α signaling only yielded moderate inhibition of adipogenesis. Here we showed that in addition to promoting the TNF-α signaling, alcohol also suppressed the Wnt1/β-catenin signaling pathway. Methods: We treated primary BMSCs from human subjects with alcohol for 24 days. We measured changes of genes related to endoplasmic reticulum (ER) stress, adipogenic markers and osteogenic markers using quantitative real-time RT-PCR and Western blot analysis. We performed Alizarin red staining for osteogenesis. We also conducted assays for osteogenic biomarkers alkaline phosphatase, collagen-I and osteocalcin. Results: Wnt/β-catenin signaling was markedly activated in BMSCs treated with osteogenic inducers relative to the control cells, as indicated by the increased levels of nuclear β-catenin along with reduced levels of cytosolic β-catenin, as well as increased protein levels of Wnt1. Activation of Wnt/β-catenin signaling was significantly suppressed in BMSCs exposed to alcohol, which was reflected by downregulated expression of osteogenic marker genes Osf2/Cbfa1, osteopontin and osteocalcin, upregulated adipogenic marker PPARγ2 and aP2, and reduced number of calcifcation nodules. In contrast, activation of Wnt/β-catenin signaling by BIO favored osteogenesis even in the presence of alcohol. Simultaneous activation of Wnt1 by BIO and inhibition of TNF-α by 3,6'-dithiothalidomide produced synergetic suppression of ethanol-induced adipogenic lineage compared to interference with either of them alone. Conclusion: This remarkable shift of BMSCs towards osteoblast lineage suggests the superiority of concordant and reciprocal interferences of the TNF-α and Wnt/β-catenin pathways for promoting osteogenesis. |
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| institution | Directory Open Access Journal |
| issn | 1015-8987 1421-9778 |
| language | English |
| last_indexed | 2024-04-12T06:01:33Z |
| publishDate | 2013-09-01 |
| publisher | Cell Physiol Biochem Press GmbH & Co KG |
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| series | Cellular Physiology and Biochemistry |
| spelling | doaj.art-5ca4e0a56a664509a253bfbd42997e942022-12-22T03:45:00ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782013-09-0132375576510.1159/000354477354477Reciprocal Interferences of TNF-α and Wnt1/β-Catenin Signaling Axes Shift Bone Marrow-Derived Stem Cells Towards Osteoblast Lineage after Ethanol ExposureYueping ChenLiang ChenQingshui YinHui GaoPanfeng DongXiaoyun ZhangJie KangBackground/Aims: We have reported in a separate study that alcohol exposure triggers activation of the TNF-α signaling pathway leading to an adverse shift of multipotential mesenchymal stem cells in bone marrow (BMSCs) away from osteogenesis towards adipogenesis. However, inhibition of TNF-α signaling only yielded moderate inhibition of adipogenesis. Here we showed that in addition to promoting the TNF-α signaling, alcohol also suppressed the Wnt1/β-catenin signaling pathway. Methods: We treated primary BMSCs from human subjects with alcohol for 24 days. We measured changes of genes related to endoplasmic reticulum (ER) stress, adipogenic markers and osteogenic markers using quantitative real-time RT-PCR and Western blot analysis. We performed Alizarin red staining for osteogenesis. We also conducted assays for osteogenic biomarkers alkaline phosphatase, collagen-I and osteocalcin. Results: Wnt/β-catenin signaling was markedly activated in BMSCs treated with osteogenic inducers relative to the control cells, as indicated by the increased levels of nuclear β-catenin along with reduced levels of cytosolic β-catenin, as well as increased protein levels of Wnt1. Activation of Wnt/β-catenin signaling was significantly suppressed in BMSCs exposed to alcohol, which was reflected by downregulated expression of osteogenic marker genes Osf2/Cbfa1, osteopontin and osteocalcin, upregulated adipogenic marker PPARγ2 and aP2, and reduced number of calcifcation nodules. In contrast, activation of Wnt/β-catenin signaling by BIO favored osteogenesis even in the presence of alcohol. Simultaneous activation of Wnt1 by BIO and inhibition of TNF-α by 3,6'-dithiothalidomide produced synergetic suppression of ethanol-induced adipogenic lineage compared to interference with either of them alone. Conclusion: This remarkable shift of BMSCs towards osteoblast lineage suggests the superiority of concordant and reciprocal interferences of the TNF-α and Wnt/β-catenin pathways for promoting osteogenesis.http://www.karger.com/Article/FullText/354477EthanolMultipotential mesenchymal stem cells (MSC)Bone-marrow stem cells (BMSC)OsteoblastOsteogenesisAdipogenisisTNF-αWnt1β-cateninsiRNA |
| spellingShingle | Yueping Chen Liang Chen Qingshui Yin Hui Gao Panfeng Dong Xiaoyun Zhang Jie Kang Reciprocal Interferences of TNF-α and Wnt1/β-Catenin Signaling Axes Shift Bone Marrow-Derived Stem Cells Towards Osteoblast Lineage after Ethanol Exposure Cellular Physiology and Biochemistry Ethanol Multipotential mesenchymal stem cells (MSC) Bone-marrow stem cells (BMSC) Osteoblast Osteogenesis Adipogenisis TNF-α Wnt1 β-catenin siRNA |
| title | Reciprocal Interferences of TNF-α and Wnt1/β-Catenin Signaling Axes Shift Bone Marrow-Derived Stem Cells Towards Osteoblast Lineage after Ethanol Exposure |
| title_full | Reciprocal Interferences of TNF-α and Wnt1/β-Catenin Signaling Axes Shift Bone Marrow-Derived Stem Cells Towards Osteoblast Lineage after Ethanol Exposure |
| title_fullStr | Reciprocal Interferences of TNF-α and Wnt1/β-Catenin Signaling Axes Shift Bone Marrow-Derived Stem Cells Towards Osteoblast Lineage after Ethanol Exposure |
| title_full_unstemmed | Reciprocal Interferences of TNF-α and Wnt1/β-Catenin Signaling Axes Shift Bone Marrow-Derived Stem Cells Towards Osteoblast Lineage after Ethanol Exposure |
| title_short | Reciprocal Interferences of TNF-α and Wnt1/β-Catenin Signaling Axes Shift Bone Marrow-Derived Stem Cells Towards Osteoblast Lineage after Ethanol Exposure |
| title_sort | reciprocal interferences of tnf α and wnt1 β catenin signaling axes shift bone marrow derived stem cells towards osteoblast lineage after ethanol exposure |
| topic | Ethanol Multipotential mesenchymal stem cells (MSC) Bone-marrow stem cells (BMSC) Osteoblast Osteogenesis Adipogenisis TNF-α Wnt1 β-catenin siRNA |
| url | http://www.karger.com/Article/FullText/354477 |
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