Isorhamnetin activates p38 signaling to promote osteogenic differentiation of murine mesenchymal stem cells

doi:10.3877/cma.j.issn.1674-134X.2021.04.007

Abstract

Abstract:

Objective

To investigate the effects of isorhamnetin on osteogenic differentiation of bone marrow mesenchymal stem cells.

Methods

The bone marrow mesenchymal stem cells of three-week-old C57BL/6J mouse were induced to osteogenic differentiation under osteogenic-medium. The effects of isorhamnetin among different concentrations on osteoblast toxicity were detected by cell counting kit-8 reagent and analyzed by t test for the lowest effective concentration. Alkaline phosphatase (ALP) and alizarin red (AR) staining kits were used to observe the effects of isorhamnetin on osteogenic differentiation and mineralization at effective concentrations. Real time (RT)-PCR was used to detect the expression of osteoblast-related genes, including osteopontin (OPN), osteocalcin (OCN), and ALP, the data were analyzed by t test. Western blot and immunofluorescence were used to confirm that isorhamnetin promoted osteogenic differentiation-related pathways.

Results

The isorhamnetin concentrations of 5 μmol/L (t=5.4, P<0.01) and 10 μmol/L (t=7.2, P<0.01) promoted the proliferation of bone marrow mesenchymal stem cells, while isorhamnetin of 20 μmol/L (t=4.2, P<0.01) significantly inhibited cell proliferation and exhibited toxic effects. The ALP and AR staining revealed that 10 μmol/L isorhamnetin significantly promoted osteogenic differentiation and mineralization compared with the control group. It was found that isorhamnetin activated the p38 signaling pathway.

Conclusion

The isorhamnetin of 10 μmol/L promotes osteogenic differentiation of bone marrow mesenchymal stem cells by activating the p38 signaling pathway.

Key words: Osteoblast, Signal transduction, Mesenchymal stem cells

Jun Fan, Liping Cao. Isorhamnetin activates p38 signaling to promote osteogenic differentiation of murine mesenchymal stem cells[J]. Chinese Journal of Joint Surgery(Electronic Edition), 2021, 15(04): 432-437.

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