Enhancement of osteogenic performance in bone marrow mesenchymal stem cells through corynoline-mediated regulation of macrophage polarization
LI Rou
LI Jing
GUO Yanning
ZHANG Kai
XU Shiqing
JING Yanan
CAO Cong
HE Xiaotong
XU Baohua
Abstract：Objective To investigate whether corynoline enhances the migration and osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)through the regulation of macrophage polarization.Methods The CCK-8 assay was employed to determine the optimal experimental concentration of corynoline.RAW264.7 cells were induced with lipopolysaccharide(LPS)to polarize into the M1 phenotype.The study included a control group(0 μmol/L)and three experimental groups treated with low,medium,and high concentrations of corydaline(1,4,and 8 μmol/L,respectively).The expression of inflammation-related indicators was determined by flow cytometry,immunofluorescence,qRT-PCR and Western blot.The reactive oxygen species(ROS)experiment was used to detect the changes in ROS content.Cell scratch experiments were conducted in the scratch-control group,scratch-Cor group,scratch-LPS group,and scratch-LPS+Cor group to observe the cell migration in each group.Osteogenic differentiation was detected in the osteogenesis-control group,osteogenesis-Cor group,osteogenesis-LPS group and osteogenesis-LPS+Cor group to observe the content of alkaline phosphatase(ALP).Results Flow cytometry and immunofluorescence detection showed that the expression of CD86 gradually decreased in the control group,low-concentration group,medium-concentration group and high-concentration group,while the expression of CD206 gradually increased.qRT-PCR detection and Western blot detection showed that the expressions of M1-related genes tumor necrosis factor-α and interleukin-1 were gradually down-regulated in the control group,low-concentration group,medium-concentration group and high-concentration group,while the expressions of M2-related genes arginase 1 and interleukin-10 were gradually up-regulated.Moreover,compared with the control group,the changes of M1 down-regulation and M2 up-regulation in the high-concentration group were the most significant(P<0.01).The ROS experiment showed that the fluorescence intensity of DCFH-DA in cells gradually decreased in the low-concentration group,medium-concentration group and high-concentration group,with the most significant decrease in the high-concentration group(P<0.001).The cell scratch assay showed that the cell migration rates of the scratch-LPS+Cor group,the scratch-LPS group,the scratch-Cor group,and the scratch-control group decreased successively.ALP staining and quantitative analysis showed that at 14 days of osteogenic culture,the ALP activity in the osteogenesis-LPS+Cor group was significantly higher than that in the osteogenesis-LPS group,the osteogenesis-Cor group,and the osteogenesis-control group(P<0.001).Conclusion A high concentration(8 μmol/L)of corynoline effectively inhibits the polarization of RAW264.7 cells toward the M1 phenotype,promotes their differentiation into the M2 phenotype,and significantly enhances the mediation of RAW264.7 cell polarization to facilitate BMSC migration and osteogenic differentiation.These findings provide a novel insight into the anti-inflammatory mechanism of corynoline and its potential therapeutic application in bone injury-related diseases.
Keywords：corynolineinflammationmacrophagepolarizationbone marrow mesenchymal stem cellosteogenic differentiation
Publication Date：2025-11-20
Online Publishing Date：2025-12-02(First online date of this platform, not the publication date of the document)
Pages：7( 406-412 )