International Journal of Stem Cells

Korean Society for Stem Cell Research (한국줄기세포학회)
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Wedelolactone Promotes the Chondrogenic Differentiation of Mesenchymal Stem Cells by Suppressing EZH2

  • Wei Qin (Research Center for Integrative Medicine, School of Basic Medical Sciences, Guangzhou University of Chinese Medicine) ;
  • Lin Yang (Traditional Chinese Medicine Innovation Research Center, Shenzhen Hospital of Integrated Traditional Chinese and Western Medicine) ;
  • Xiaotong Chen (Guangdong Key Laboratory of Liver Disease Research, The Third Affiliated Hospital of Sun Yat-Sen University) ;
  • Shanyu Ye (Research Center for Integrative Medicine, School of Basic Medical Sciences, Guangzhou University of Chinese Medicine) ;
  • Aijun Liu (Research Center for Integrative Medicine, School of Basic Medical Sciences, Guangzhou University of Chinese Medicine) ;
  • Dongfeng Chen (Research Center for Integrative Medicine, School of Basic Medical Sciences, Guangzhou University of Chinese Medicine) ;
  • Kunhua Hu (Guangdong Key Laboratory of Liver Disease Research, The Third Affiliated Hospital of Sun Yat-Sen University)
  • Received : 2022.03.18
  • Accepted : 2022.09.01
  • Published : 2023.08.30
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Abstract

Background and Objectives: Osteoarthritis (OA) is a degenerative disease that leads to the progressive destruction of articular cartilage. Current clinical therapeutic strategies are moderately effective at relieving OA-associated pain but cannot induce chondrocyte differentiation or achieve cartilage regeneration. We investigated the ability of wedelolactone, a biologically active natural product that occurs in Eclipta alba (false daisy), to promote chondrogenic differentiation. Methods and Results: Real-time reverse transcription-polymerase chain reaction, immunohistochemical staining, and immunofluorescence staining assays were used to evaluate the effects of wedelolactone on the chondrogenic differentiation of mesenchymal stem cells (MSCs). RNA sequencing, microRNA (miRNA) sequencing, and isobaric tags for relative and absolute quantitation analyses were performed to explore the mechanism by which wedelolactone promotes the chondrogenic differentiation of MSCs. We found that wedelolactone facilitates the chondrogenic differentiation of human induced pluripotent stem cell-derived MSCs and rat bone-marrow MSCs. Moreover, the forkhead box O (FOXO) signaling pathway was upregulated by wedelolactone during chondrogenic differentiation, and a FOXO1 inhibitor attenuated the effect of wedelolactone on chondrocyte differentiation. We determined that wedelolactone reduces enhancer of zeste homolog 2 (EZH2)-mediated histone H3 lysine 27 trimethylation of the promoter region of FOXO1 to upregulate its transcription. Additionally, we found that wedelolactone represses miR-1271-5p expression, and that miR-1271-5p post-transcriptionally suppresses the expression of FOXO1 that is dependent on the binding of miR-1271-5p to the FOXO1 3'-untranscribed region. Conclusions: These results indicate that wedelolactone suppresses the activity of EZH2 to facilitate the chondrogenic differentiation of MSCs by activating the FOXO1 signaling pathway. Wedelolactone may therefore improve cartilage regeneration in diseases characterized by inflammatory tissue destruction, such as OA.

Keywords

Acknowledgement

We would like to thank Dr. Wei Zhao for valuable suggestions. We also thank all of the students who participated in this study. This work was supported by Guangdong Innovative and Entrepreneurial Research Team Program (2016ZT06S029), the Natural Science Foundation of Guangdong Province (2017A030312009) and the China Postdoctoral Science Foundation (2021M700961).

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