Periodic Mechanical Stress Stimulates the FAK Mitogenic Signal in Rat Chondrocytes Through ERK1/2 Activity

Background/Aims: The biological effects of periodic mechanical stress on chondrocytes have been studied extensively over the past few years. However, the mechanisms underlying chondrocyte mechanosensing and signaling in response to periodic mechanical stress remain to be determined. In the current study, we examined the effects of focal adhesion kinase (FAK) signaling on periodic mechanical stress-induced chondrocyte proliferation and matrix synthesis. Methods and Results: Periodic mechanical stress significantly induced sustained phosphorylation of FAK at Tyr397 and Tyr576/577. Reduction of FAK with targeted shRNA via transfection of NH2-terminal tyrosine phosphorylation-deficient FAK mutant Y397F or Y576FY577F abolished periodic mechanical stress-induced chondrocyte proliferation and matrix synthesis, accompanied by attenuated ERK1/2 phosphorylation. However, activation of Src, PLCyl and Racl was not prevented upon FAK suppression. Furthermore, pretreatment with the Src-selective inhibitor, PP2, and shRNA targeted to Src or suppression of Racl with its selective inhibitor, NSC23766, blocked FAK phosphorylation at Tyr,5767577 but not Tyr,397 under periodic mechanical stress. Interestingly, FAK phosphorylation neither at Tyr397 nor at Tyr5761577 was affected by PLCyl depletion when periodic mechanical stress was applied. In addition, Tyr397 and Tyr5761577 phosphorylation levels were reduced upon pretreatment with a blocking antibody against integrin 31 under conditions of periodic mechanical stress. Conclusion: Our findings collectively suggest that periodic mechanical stress promotes chondrocyte proliferation and matrix synthesis through at least two pathways, integrin 131-Src-Racl-FAK(Tyr5761577)-ERK1/2 and integrin ill-FAK (Tyr397)-ERK1/2. Copyright (C) 2013 S. Karger AG, Basel