Effect And Mechanism Of Sox9 Targeting Wnt/β-catenin Pathway On Chondrogenic Differentiation Of Adipose Stem Cells

Objective To study the effect of SRY-box containing protein 9(SOX9)on chondrogenic differentiation of adipose stem cells(ADSCs),and to explore the possible mechanism.Methods ADSCs cells were divided into blank group,no-load group,overexpression group and combination group.The blank group is not processed.The noload group was transfected with pc DNA3.1(+)-NC plasmid,the overexpression group was transfected with pc DNA3.1(+)-Sox9 plasmid,and the combined group was transfected with pc DNA3.1(+)-Sox9 plasmid,and Li Cl was added to make its final concentration 60mmol/L.48 h after transfection,the protein expressions of Sox9,β-catenin(β-catenin),p-β-catenin and glycogen synthase kinase 3β(GSK-3β)were detected by Western blot.The proliferation activity of ADSCs was detected by MIT assay.ADSCs were induced to differentiate into chondrocytes,and the expression of Aggrecan was observed by toluidine blue staining after 21 days.The protein expression of collagen type II(Collagen II)was detected by Western blot.Results Compared with the blank group and the no-load group,the A of the overexpression group were increased at 24,48,and 72 h in the MTT experiment(P<0.05).Compared with the overexpression group,the A of the combination group were decreased at 24,48,and 72 h in the MTT experiment(P<0.05).Toluidine blue staining showed that both the cell membrane and cytoplasm were stained blue.The staining intensity of the overexpression group was stronger than that of the combination group,and the combination group was stronger than the blank group and the empty group.There was no significant difference in the staining intensity between the blank group and the empty group.The expression group expressed a large amount of Aggrecan.Compared with the blank group and no-load group,the expression of Collagen II protein in the overexpression group was increased(P<0.05).Compared with the overexpression group,the expression of Collagen II protein in the combination group was decreased(P<0.05).Conclusion Overexpression of Sox9 can promote the proliferation and cartilage differentiation of ADSCs,and its mechanism may be related to the inhibition of Wnt/β-catenin pathway.