Melatonin contributes to the hypertrophic differentiation of mesenchymal stem cell-derived chondrocytes via activation of the Wnt/β-catenin signaling pathway

Abstract Background Hypertrophy is a critical process for chondrocyte differentiation and maturation during endochondral ossification, which is responsible for the formation of long bone and postnatal longitudinal growth. Increasing evidence suggests that melatonin, an indole hormone, plays a pivotal role in chondrogenesis. However, little is known about the effects of melatonin on the terminal differentiation of chondrocytes. Methods Mesenchymal stem cell (MSC)-derived chondrocytes generated by a high-density micromass culture system were induced to undergo hypertrophic differentiation. Melatonin-mediated hypertrophic differentiation was examined by reverse transcription polymerase chain reaction analysis (RT-PCR) analysis, histological staining and immunohistochemistry. Activation of the Wnt signaling pathway was evaluated by PCR array, RT-PCR, western blotting and immunofluorescence. XAV-939, a Wnt signaling pathway antagonist, was further used to determine whether the effect of melatonin on chondrocyte hypertrophic differentiation was mediated occurred by activation of Wnt signaling pathway. Results Histological staining showed melatonin increased chondrocyte cell volume and the expression of type X collagen but decreased the expression of type II collagen compared with the control group. RT-PCR showed that melatonin significantly up-regulated the gene expressions of biomarkers of hypertrophic chondrocytes, including type X collagen, alkaline phosphatase, runt-related transcription factor 2, Indian hedgehog and parathyroid hormone-related protein receptor, and melatonin down-regulated the mRNA expression of hallmarks of chondrocytes, including parathyroid hormone-related protein. PCR array showed that the effect of melatonin on chondrocyte hypertrophic differentiation was accompanied by the up-regulation of multiple target genes of the canonical Wnt signaling pathway, and this effect was blocked by XAV-939. Conclusions The current findings demonstrate that melatonin enhances the hypertrophic differentiation of MSC-derived chondrocytes through the Wnt signaling pathway. Our findings add evidence to the role of melatonin in promoting bone development and highlight the positive effects of melatonin on terminal differentiation of chondrocytes.

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Melatonin Contributes to The Hypertrophic Differentiation of Mesenchymal Stem Cell-Derived Chondrocytes Via Activation of The WNT/β-Catenin Signaling Pathway

10.21203/rs.3.rs-448728/v1 ◽

2021 ◽

Author(s):

Xuan Wang ◽

Tianwei He ◽

Lei He ◽

Bu Yang ◽

Zhongyu Liu ◽

...

Keyword(s):

Parathyroid Hormone ◽

Wnt Signaling ◽

Signaling Pathway ◽

Wnt Signaling Pathway ◽

Terminal Differentiation ◽

Related Protein ◽

Rt Pcr ◽

Type X Collagen ◽

Parathyroid Hormone Related Protein ◽

Histological Staining

Abstract BackgroundHypertrophy is a critical process for chondrocyte differentiation and maturation during endochondral ossification, which is responsible for the formation of long bone and its postnatal longitudinal growth. Increasing evidence suggests that melatonin, an indole hormone, plays a pivotal role in chondrogenesis; however, little is known about its effects on the terminal differentiation of chondrocytes. MethodsMesenchymal stem cells (MSCs) derived chondrocytes generated by a high-density micromass culture system were further induced to hypertrophic differentiation. The melatonin-mediated hypertrophic differentiation were examined by reverse transcription polymerase chain reaction analysis (RT-PCR) analysis and histological staining and immunohistochemistry. The expression of downstream factors of WNT signaling pathway was evaluated by RT-PCR, western blotting and immunofluorescence. The WNT signaling pathway antagonist XAV-939 was used to further demonstrate melatonin-induced hypertrophic chondrocytes and WNT signaling pathway activation.ResultsHistological staining showed melatonin increased the chondrocytes cell volume and the expression of type X collagen, but decreased the expression of type II collagen compare to the control group. RT-PCR showed that melatonin signiﬁcantly up-regulated the expression of biomarkers of hypertrophic chondrocytes, including type X collagen (COL10A1), alkaline phosphatase (ALP), Runt-related transcription factor 2 (RUNX2), Indian hedgehog (IHH), and parathyroid hormone-related protein receptor (PTHrP-R) and down-regulated the hallmarks of chondrocytes, including parathyroid hormone-related protein (PTHrP). The WNT signaling pathway PCR array showed that the effect of melatonin was accompanied by the up-regulation of multiple target genes of the canonical WNT signaling pathway and the melatonin-mediated effect can be blocked by XAV-939. ConclusionsThese ﬁndings demonstrate that melatonin can enhance the hypertrophic differentiation of MSCs-derived chondrocytes through the WNT signaling pathway. It adds evidence to the role of melatonin in promoting bone development and highlights its positive effects on chondrocytes terminal differentiation.

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Secreted frizzled related protein 1 protects H9C2 cells from hypoxia/re-oxygenation injury by blocking the Wnt signaling pathway

Lipids in Health and Disease ◽

10.1186/s12944-016-0240-5 ◽

2016 ◽

Vol 15 (1) ◽

Cited By ~ 13

Author(s):

Jing Tao ◽

Mayila Abudoukelimu ◽

Yi-tong Ma ◽

Yi-ning Yang ◽

Xiao-mei Li ◽

...

Keyword(s):

Wnt Signaling ◽

Signaling Pathway ◽

Wnt Signaling Pathway ◽

H9c2 Cells ◽

Related Protein

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Reverse Regulation Between sFRP 5 and Dkk 1 Gene in Early Stage Lung Adenocancer

10.21203/rs.3.rs-434159/v1 ◽

2021 ◽

Author(s):

Arife Zeybek ◽

Necdet Oz ◽

Serdar Kalemci ◽

Kursad Tosun ◽

Tuba Gökdoğan Edgünlü ◽

...

Keyword(s):

Lung Adenocarcinoma ◽

Wnt Signaling ◽

Signaling Pathway ◽

Tumor Tissue ◽

Early Stage ◽

Wnt Signaling Pathway ◽

Normal Lung ◽

Pcr Analysis ◽

Related Protein ◽

Expression Levels

Abstract Purpose: We aimed to examine the expression levels of the genes of APC (Adenomatous Polyposis Coli) 1, APC 2, Dkk (Dickkopf related protein) 1, Dkk -3, sFRP (Secreted frizzled-related protein) -2, sFRP-4, and sFRP-5 genes which play a role in the Wnt signaling pathway in lung adenocarcinoma and adjacent normal lung tissues, and to evaluate their relationship with clinical-pathological factors.Materials and methods: Between 2011 and 2018, the expression levels of the relevant genes in formalin-fixed paraffin-embedded tumor and adjacent intact lung tissue samples of 57 patients who were operated for lung adenocarcinoma were determined by Real-time PCR analysis. Results: The expression levels of the Dkk-1 gene in the tumor tissue, especially in stage I-II, were statistically significantly suppressed compared to normal tissue (p <0.025 ). Although Dkk-1 gene expression was suppressed in the tumor tissue of patients with early-stage lung adenocarcinoma, the level of expression of the sFRP-5 gene was found to be statistically significantly higher (p<0.039). Conclusion: In our study, between the sFRP-5 and Dkk-1 genes, known as the extracellular antagonist of the Wnt signaling pathway was found the reverse regulation. sFRP-5 gene was found as having an oncogenic role in adenocarcinoma development. Reverse regulation between these genes in early-stage lung adenocarcinoma may shed light on the mechanisms associated with the development of carcinogenesis. For that reason, clinically, this relationship needs to research in a larger series of pure adenocarcinoma and normal human lung tissues, separated by its stage, for potential therapeutic target or prognostic its significance.

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