Effects Of AKT Signaling Pathway On The Cartilage Degeneration Of The Endplate Under Intermittent Cyclic Mechanical Tension

Objective: On the basis of cell culture in vitro,establish the model of cell degeneration under intermittent cyclic mechanical tension.To investigate the effect of AKT/m TOR signaling pathway on the degeneration of cartilage in the endplate under intermittent cyclic mechanical tension.Methods: Isolate and culture rat endplate chondrocytes,and to establish a model for the degeneration of the cartilage cells under FX5000 T.Divided into blank control group(P2 cells);experimental group 1:FX5000T intermittent cyclic mechanical tension.;experimental group 2:FX5000T intermittent cyclic mechanical tension and AKT/m TOR signal pathway inhibitor group;inverted phase contrast microscope was used to observe the morphology of endplate cartilage cells of each group;toluidine blue staining to detect the endplate chondrocytes proteoglycan changes;Western blotting was used to detect AKT,P-AKT and protein expression of AKT in the downstream;Used real time PCR to detect the expression of the SOX-9,type ?collagen,Aggrecan and cell matrix metabolism genes MMP-3,MMP-13,Adamts-5and other related gene expressionResults: With FX5000 T stress loading,rat endplate chondrocytes in cartilage markers:SOX9,type ? collagen,aggrecan gene expression gradually decreased and then gradually increased;That at the beginning of the endplate chondrocytes of rat in the extracellular matrix synthesis in the beginning were inhibited.Under the effect of intermittent cyclic mechanical tension and endplate chondrocytes of rat chondrocyte gene symbol and Akt / m TOR signaling pathway is the key of gene and protein expression was decreased gradually;the inhibition of Akt / m TOR signaling pathway,rat chondrocytes gene marker expression was significantly decreased,Akt / m TOR signaling pathway related gene and protein expression is greatly reduced,and the prompt rat endplate chondrocyte degeneration was significantly increased.Conclusion: Intermittent cyclic mechanical tension through the activation of AKT/m TOR/P70S6 K cell signaling pathways,induce endplate cartilage cells of rats in vitro degeneration model.It is play an important role under the action of the tension of the degeneration process.Artificial control or intervention AKTm TORP70S6 K expression of signaling pathways may slow down,or inhibition of endplate cartilage degeneration in rats.