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  

The role of mechanical stress combined with verapamil on aggrecan metabolism in osteoarthritic chondrocytes-agarose constructs

作　　者：罗丹[1] 申艳[1] 闾坚强[2] 范永前[3] 黄东辉[3] 林伟龙[3] 沈海敏[3] 徐沪济[4] 管剑龙[5] 

机构地区：[1]复旦大学附属华东医院免疫风湿科科,上海200040 [2]上海体育学院运动科学学院,上海200433 [3]复旦大学附属华东医院骨科,上海200040 [4]第二军医大学长征医院风湿免疫科,上海200003 [5]复旦大学附属华东医院免疫风湿科,上海200040

出　　处：《复旦学报（医学版）》2016年第3期262-267,共6页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金面上项目(81072478,81273298)~~

摘　　要：目的通过对骨关节炎软骨细胞-琼脂糖施压模型机械施压及联合维拉帕米干预,研究软骨细胞聚集蛋白聚糖(aggrecan,AGC)代谢的变化。方法选取膝关节炎患者关节置换术后胫骨平台软骨,分离软骨细胞,制作软骨细胞-琼脂糖三维模型,设置空白对照组、施压组和施压联合维拉帕米组。采用Flexcell FX-5000TM细胞加力仪施压(24 kPa,0.33 Hz,2 h),维拉帕米干预浓度设定为40μmol/L。采用RT-PCR法和ELISA法分别检测AGC、ADAMTS-4、ADAMTS-5 mRNA和蛋白质水平。结果与空白对照组比较,施压组AGC mRNA水平显著升高(P=0.000),ADAMTS-5 mRNA水平显著降低(P=0.000),施压联合维拉帕米组AGC mRNA水平显著升高(P=0.012),ADAMTS-4和ADAMTS-5 mRNA水平均显著降低(P=0.000)。与空白对照组比较,施压组AGC mRNA水平显著增高(P=0.000)、ADAMTS-5水平显著降低(P=0.000);与施压组比较,施压联合维拉帕米组AGC水平显著增高(P=0.008),ADAMTS-4和ADAMTS-5水平均显著降低(P=0.000)。结论机械应力可促进骨关节炎软骨细胞AGC合成,维拉帕米可以加强机械应力的作用。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-5000 TMwas applied to impose cyclic mechanical stress（24kPa,0.33 Hz,2 h）,and verapamil（40μmol/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-5at 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.

关 键 词：骨关节炎 软骨细胞 蛋白聚糖 机械应力 维拉帕米 

分 类 号：R681.3[医药卫生—骨科学]