The Role Of AhR In Osteogenic Differentiation Of Induced Pluripotent Stem Cells And The Intervention Of Psoralea Corylifolia Linn.

Objective: Io explore the role of aryl hydrocarbon receptor(AhR)in the osteogenic differentiation of iPSCs,and to provide a theoretical basis for screening iPSCs osteogenic differentiation inducers by targeting AhR.Ihe role of psoralea corylifolia Linn.and its active component isopsoralen in osteogenic differentiation of iPSCs and their effect on AhR were also investigated,which provided an experimental basis for the induction of osteogenic differentiation of iPSCs in vitro with Chinese medicine or its active component targeting AhR.Methods: Ihe osteogenic differentiation of iPSCs was induced by adding adenosine,ascorbic acid and ?-glycerophosphate.Ihe expression trend of AhR at different time points of iPSCs osteogenic differentiation was detected.AhR agonists and antagonists were used to investigate the role of AhR in osteogenic differentiation of iPSCs,and the effect of AhR agonists on the expression of estrogen receptor ?(ER?)was also observed.Psoralea corylifolia Linn.and its active component isopsoralen were used to intervene the osteogenic differentiation of iPSCs,and their roles in the osteogenic differentiation of iPSCs were observed.Ihe targets of psoralea corylifolia Linn.for promoting osteogenic differentiation were predicted based on network pharmacology and verified by experiments.Ihe effect of isopsoralen on AhR and its target gene expression was detected,and the binding of isopsoralen with AhR was discussed based on the drug affinity responsive target stability.Results:(1)Ihe expression of pluripotent genes and osteogenic genes and the results of alizarin red staining showed that the method of directly inducing osteogenic differentiation of iPSCs was successfully established.(2)AhR expression was downregulated at different time points of iPSCs osteogenic diffeentiation,and its agonist inhibited osteogenic differentiation,while its antagonist promoted osteogenic differentiation,and AhR agonist reduced ER? protein level.(3)Psoralea corylifolia Linn.could promote osteogenic differentiation of iPSCs.Network pharmacology predicted that psoralea corylifolia Linn.had 71 targets that contributed to bone action.AhR and peroxisome proliferator activated receptor ?(PPAR?)were verified.(4)Isopsoralen could also promote osteogenic differentiation of iPSCs and inhibit the expression of AhR and its target genes.Conclusion: In this study,we established a direct method to induce osteogenic differentiation of iPSCs.AhR played a negative role in the osteogenic differentiation of iPSCs,and may contributed to bone formation by regulating ER? expression.Psoralea corylifolia Linn.and its active component isopsoralen could promote osteogenic differentiation of iPSCs by inhibiting nuclear AhR expression.