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Small Molecule Nitazoxanide Inhibits Osteogenic Differentiation and Promotes Adipogenic Differentiation of Bone
Objective:
To investigate the potential effect of small molecule nitazoxanide (NTZ) on the osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).
Methods:
Cell counting Kit-8 assay was used to examine the effect of NTZ on proliferation of BMSCs. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot analysis were used to measure the expression of osteogenic and adipogenic marker gene. Alkaline phosphatase (ALP) staining and activity assay and Alizarin Red S (ARS) staining were used to investigate the effect of NTZ on osteogenesis. Oil red O (ORO) staining assay was used to assess the impact of NTZ on adipogenesis.
Results:
NTZ significantly suppressed the osteogenic differentiation but promoted the adipogenic differentiation of BMSCs. Mechanistically, NTZ regulated osteogenic/adipogenic differentiation of BMSCs by inhibiting the Wnt/β-catenin signalling pathway. The addition of Wnt/β-catenin signalling pathway activator, lithium chloride, could reverse the effect of NTZ on BMSCs.
Conclusion:
NTZ affected osteogenic and adipogenic differentiation of BMSCs with the involvement of Wnt/β-catenin signalling pathway. This finding expanded the understanding of NTZ pharmacology and indicated that NTZ might have an adverse effect on bone homeostasis.
Insights
Nitazoxanide (NTZ) suppresses bone formation and promotes fat formation in stem cells by inhibiting the Wnt/β-catenin pathway. This suggests NTZ may negatively impact bone health.
Area of Science:
- Stem cell biology
- Pharmacology
- Bone biology
Background:
- Bone marrow mesenchymal stem cells (BMSCs) differentiate into osteoblasts (bone-forming cells) or adipocytes (fat-forming cells).
- The balance between osteogenic and adipogenic differentiation is crucial for maintaining bone homeostasis.
- Nitazoxanide (NTZ) is a small molecule with known antiparasitic and antiviral properties.
Purpose of the Study:
- To investigate the effect of nitazoxanide (NTZ) on the osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).
Main Methods:
- Cell proliferation was assessed using the Cell Counting Kit-8 assay.
- Gene and protein expression of osteogenic and adipogenic markers were analyzed by qRT-PCR and Western blot.
- Osteogenesis was evaluated using alkaline phosphatase (ALP) staining and activity assays, and Alizarin Red S (ARS) staining.
- Adipogenesis was assessed using Oil Red O (ORO) staining.
Main Results:
- NTZ significantly suppressed osteogenic differentiation of BMSCs.
- NTZ promoted adipogenic differentiation of BMSCs.
- NTZ inhibited the Wnt/β-catenin signaling pathway, which was reversed by lithium chloride, a Wnt/β-catenin activator.
Conclusions:
- NTZ influences BMSC differentiation towards adipogenesis and away from osteogenesis.
- The Wnt/β-catenin signaling pathway is involved in NTZ's effects on BMSC differentiation.
- NTZ may adversely affect bone homeostasis due to its impact on BMSC differentiation.
