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Interleukin-35 Promote Osteogenesis and Inhibit Adipogenesis: Role of Wnt/β-Catenin and PPARγ Signaling Pathways

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Abstract

Mesenchymal stem cells (MSCs) are multipotent stem cells that are able to differentiate into several cell types, including cartilage, fat, and bone. It has been reported that the decision process of MSCs into fat and bone cells is competing and reciprocal. Interleukin (IL)-35 is an important effector protein in the Wnt/β-catenin signaling pathway that acts as a bone metabolism regulator. However, it is unclear whether IL-35 is also important for regulating MSC differentiation to fat and bone. In the current study, we evaluated the role of IL-35 in C3H10T1/2 cells, which are a good cell model for investigating osteogenesis and adipogenesis in bone marrows. The role of IL-35 on osteoblast proliferation and apoptosis was assessed using cell counting kit-8 assay and flow cytometry, respectively. Extracellular matrix mineralization and lipid accumulation were measured by Alizarin Red S staining and Oil Red O staining, respectively. The most important transcription factor of the process of osteogenesis Runx2 and Wnt/β-catenin signaling pathway components β-catenin and Axin2 were investigated in response to IL-35 treatment. Furthermore, the adipogenic markers PPAR-γ and C/EBPα were also investigated. Our observations showed that IL-35 could promote the proliferation of MSCs and inhibit the apoptosis of MSCs. We found that IL-35 treatment resulted in a dramatic stimulation of osteogenesis and inhibition of adipogenesis. Moreover, IL-35 enhanced Wnt/β-catenin pathway key component β-catenin as well as Axin2 expression during MSCs differentiated to osteoblasts. Our findings suggested that IL-35 might control the balance between osteogenic and adipogenic differentiation of progenitor cells through the Wnt/β-catenin-PPARγ signaling pathway, suggesting its potential application in providing an intervention in osteoporosis and obesity.

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The data used to support the findings of this study are available from the corresponding author upon request.

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Funding

This study was funded by the China Postdoctoral Science Foundation (2019M661173) and the 345 Talent Project of Shengjing Hospital of China Medical University.

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Authors and Affiliations

  1. Department of Rheumatology and Immunology, Shengjing Hospital of China Medical University, No. 36 San Hao Street, Heping District, Shenyang, 110004, People’s Republic of China

    Yuxuan Li & Xiaofei Wang

  2. Department of Rheumatology and Immunology, The First Affiliated Hospital of China Medical University, 155 Nanjing North Street, Heping District, Shenyang, 110001, People’s Republic of China

    Jing Lu

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  1. Yuxuan Li
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Contributions

XF W and J.L. designed the study and revised the manuscript. YX L performed the experiments. YX L drafted the manuscript. YX L analyzed the data.

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Correspondence to Jing Lu.

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10753_2022_1749_MOESM1_ESM.tif

Supplementary file1 JASPAR website was employed to predict the binding site of Axin2 and PPARγ. The promoter sequence of Axin2 was found and the score value was greater than 80%, suggesting the transcription of Axin2 could upregulate the PPARγ expression during the MSCs differentiate to adipocytes. (TIF 8512 KB)

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Li, Y., Wang, X. & Lu, J. Interleukin-35 Promote Osteogenesis and Inhibit Adipogenesis: Role of Wnt/β-Catenin and PPARγ Signaling Pathways. Inflammation 46, 522–533 (2023). https://doi.org/10.1007/s10753-022-01749-3

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  • DOI: https://doi.org/10.1007/s10753-022-01749-3

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