Stromal Cell-Derived Factor 1α Inhibits Chondrocyte Apoptosis and Promotes Autophagy Through the Akt Signaling Pathway
Objective To investigate the effects of stromal cell-derived factor 1α(SDF-1α)on the apoptosis and autophagy of chondrocytes and the underlying mechanisms.Methods Chondrocytes were isolated from the knee joints of neonatal mice.The chondrocytes were then stimulated with 0(the control group),50,100,and 200 ng/mL of SDF-1α.CCK-8 assay was performed to determine the effects of SDF-1α stimulation for 24 h,48 h,and 72 h on the viability of the chondrocytes.Wound healing assay was conducted to determine the effects of SDF-1α stimulation for 12 h and 24 h on chondrocyte migration.The changes in the expression of Akt signaling pathway proteins in chondrocytes were determined by Western blot assay.Chondrocytes were stimulated with 0(the control group)and 200 ng/mL of SDF-1α.Flow cytometry was performed to determine the effect of SDF-1α on the apoptosis of chondrocytes.Transmission electron microscope was used to examine the effect of SDF-1α on chondrocyte autophagy.Immunofluorescence staining assays were performed to visualize the differences in p-Akt expression and distribution in chondrocytes treated with SDF-1α.Results Compared with the control group,findings for the experimental groups showed that SDF-1α at the concentrations of 50,100,and 200 ng/mL did not decrease chondrocyte activity at any time point(P<0.01)and it consistently promoted chondrocyte migration at 24 h(P<0.05).Western blot results revealed that,in comparison to the control group,SDF-1α at concentrations of 50,100,and 200 ng/mL significantly up-regulated the protein expression of p-Akt in chondrocytes,while no significant difference in Akt expression was observed.Flow cytometry demonstrated that SDF-1α could inhibit chondrocyte apoptosis(P<0.05)and transmission electron microscopic observation showed that SDF-1α promoted chondrocyte autophagy(P<0.05).Immunofluorescence staining showed that the expression of p-Akt in chondrocytes was concentrated in the perinuclear area of the cells and this expression was further enhanced in the perinuclear area of the chondrocytes after treatment with SDF-1α.Conclusion SDF-1α inhibits chondrocyte apoptosis and promotes chondrocyte migration and autophagy through activating the Akt signaling pathway.
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