Small Molecule Nitazoxanide Inhibits Osteogenic Differentiation and Promotes Adipogenic Differentiation of Bone Marrow Mesenchymal Stem Cells.

Abstract

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.