Celastrol improves self-renewal and differentiation of human tendon-derived stem cells by suppressing Smad7 through hypoxia View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2017-12

AUTHORS

Tianyi Wu, Shenghe Liu, Gen Wen, Jia Xu, Yaling Yu, Yimin Chai

ABSTRACT

BACKGROUND: We aimed to evaluate the potential enhancing effect of celastrol on the stemness of human tendon-derived stem cells (hTSCs) in vitro and the underlying molecular mechanisms. METHODS: The capability of hTSC self-renewal was assessed by cell proliferation and colony formation as determined with the CCK-8 kit. Adipogenesis, chondrogenesis, and osteogenesis were determined by Oil Red O, Alcian Blue, and Alizarin Red staining, respectively. The relative mRNA levels of Sox9, PPARγ, Runx2, Smad7, and HIF1α were determined by real-time polymerase chain reaction (PCR). The levels of Smad7 and HIF1α protein were measured by immunoblotting. The chromatin immunoprecipitation (ChIP) assay was used to assess the direct binding of HIF1α to the Smad7 promoter. Suppression of Smad7 induced by hypoxia was examined using the luciferase reporter assay. RESULTS: We found that treatment with celastrol resulted in improvement in both the multi-differentiation potential and self-renewal capability of hTSCs. Celastrol elicited hypoxia and subsequently suppressed the expression of Smad7 through direct association with the hypoxia response element consensus sequence. Further, we demonstrated that both Smad7 and HIF1α were involved in the beneficial effects of celastrol on the differentiation and self-renewal of hTSCs. CONCLUSIONS: We demonstrated the positive effect of celastrol on the stemness of hTSCs and elucidated the essential role of the HIF1α-Smad7 pathway in this process. More... »

PAGES

274

Identifiers

URI

http://scigraph.springernature.com/pub.10.1186/s13287-017-0724-x

DOI

http://dx.doi.org/10.1186/s13287-017-0724-x

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1093162898

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/29202812


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53 schema:description BACKGROUND: We aimed to evaluate the potential enhancing effect of celastrol on the stemness of human tendon-derived stem cells (hTSCs) in vitro and the underlying molecular mechanisms. METHODS: The capability of hTSC self-renewal was assessed by cell proliferation and colony formation as determined with the CCK-8 kit. Adipogenesis, chondrogenesis, and osteogenesis were determined by Oil Red O, Alcian Blue, and Alizarin Red staining, respectively. The relative mRNA levels of Sox9, PPARγ, Runx2, Smad7, and HIF1α were determined by real-time polymerase chain reaction (PCR). The levels of Smad7 and HIF1α protein were measured by immunoblotting. The chromatin immunoprecipitation (ChIP) assay was used to assess the direct binding of HIF1α to the Smad7 promoter. Suppression of Smad7 induced by hypoxia was examined using the luciferase reporter assay. RESULTS: We found that treatment with celastrol resulted in improvement in both the multi-differentiation potential and self-renewal capability of hTSCs. Celastrol elicited hypoxia and subsequently suppressed the expression of Smad7 through direct association with the hypoxia response element consensus sequence. Further, we demonstrated that both Smad7 and HIF1α were involved in the beneficial effects of celastrol on the differentiation and self-renewal of hTSCs. CONCLUSIONS: We demonstrated the positive effect of celastrol on the stemness of hTSCs and elucidated the essential role of the HIF1α-Smad7 pathway in this process.
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