机械应力联合维拉帕米对骨关节炎软骨细胞-琼脂糖三维模型蛋白聚糖代谢的作用

Objective To investigate the effect of mechanical stress and verapamil on aggrecan (AGC) metabolism by osteoarthritic chondrocytes-agarose constructs. Methods Patients with osteoarthritis were treated with knee joint replacement.Chondrocytes were isolated to construct chondrocytes-agarose model,and then were divided into placebo group,stress group and stress combined with verapamil group.Flexcell FX-5000TM was applied to impose cyclic mechanical stress (24 kPa,0.33 Hz,2 h),and verapamil (40 μmol/L) was added into stress combined with verapamil group.RT-PCR and ELISA were performed to measure the expression of AGC,ADAMTS-4 and ADAMTS-5 at both mRNA and proteins levels,respectively.Results Compared with the control group,AGC mRNA level was increased (P=0.000),and ADAMTS-5 mRNA level was significantly decreased (P=0.000) in stress alone group.Compared with the stress group,the expression of AGC mRNA was significantly elevated (P=0.012),the expressions of ADAMTS-4 mRNA (P=0.000) and ADAMTS-5 mRNA (P=0.000) were decreased in stress combined with verapamil group.Compared with the placebo group,AGC mRNA level was increased (P=0.000),and ADAMTS-5 level was significantly decreased (P=0.000) in stress group.Compared with the stress group,AGC level was increased further (P=0.008),both ADAMTS-4 and ADAMTS-5 levels were decreased (P=0.000) in stress combined with the verapamil group.Conclusions Mechanical stress can promote chondrocyte AGC synthesis,and verapamil can strengthen the role of mechanical stress.