周期性牵张应力对IL-1β刺激的人膝骨关节滑膜成纤维细胞MMPs mRNA表达的影响  

Effects of cyclic mechanical stretch on IL-1β-induced MMPs mRNA expression of synovioblast in human knee joints

作　　者：刘益杰[1] 杨松滨[1] 褚立希[1] 李信达 胡军[1] 黄正 

机构地区：[1]上海中医药大学康复医学院,上海201203 [2]中国医药大学物理治疗系,台湾台中40402 [3]上海市光华中西医结合医院关节内科,上海200052

出　　处：《上海中医药杂志》2015年第2期65-67,93,共4页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81173357);上海市卫生局医学重点专科建设计划项目(ZK2012A25);上海市教委东方学者项目(A1-N1401011325)

摘　　要：目的观察周期性牵张应力对IL-1β刺激的人膝骨关节滑膜成纤维细胞基质金属蛋白酶(MMP1、MMP3及MMP13)mRNA表达的影响。方法取第四代的人膝骨关节滑膜成纤维细胞,以每孔4×105接种于由Ⅰ型胶原蛋白包被超薄弹性硅胶膜表面的6孔Bio Flex细胞培养板,分为空白组(常规培养)、对照组(加入终浓度为10μg/L的IL-1β刺激)和牵张组(加入终浓度为10μg/L的IL-1β刺激,同时采用Flexcell细胞牵张拉伸应力加载系统对细胞进行周期性牵张力刺激),均连续干预3天;各组于牵张组实验结束后收集细胞,抽提滑膜细胞RNA,采用RT-PCR法检测细胞MMP1、MMP3和MMP13 mRNA的表达情况。结果与空白组比较,对照组MMP1、MMP3及MMP13 mRNA表达量明显升高,差异有统计学意义(P<0.05);与对照组比较,牵张组MMP1、MMP13 mRNA表达量明显下降(P<0.05),MMP3 mRNA水平下降幅度不明显(P>0.05)。结论周期性牵张应力(幅度为0-12%,频率为1 Hz,加载时间为60 min/d)能抑制人膝骨关节滑膜成纤维细胞MMP1、MMP13 mRNA的表达,从而对关节滑膜炎引起的软骨损伤起到保护作用。Objective To investigate the role of cyclic mechanical stretch on IL-1β-induced MMPs mRNA expression in synovioblasts of human knee joints.Methods The fourth-generation synovioblasts of human knee joints were seeded onto type Ⅰ collagen-coated Bio Flex 6-well culture plate at density of 4 × 10^5/well,the following three different treatment groups were devised： normal group（ untreated with IL-1β or cycle stretch）,control group（ treated with 10 ng / ml IL-1β only）,stretch group（ treated with 10 ng / ml IL-1β and subjected to cyclic mechanical stretch at a magnitude of 0-12%,frequency of 1Hz,and time of 60 min per day）. After being stretched for 3 days by Flexcell-5000 tension system,RNA was extracted and RT-PCR was performed to detect the MMP1 /3 /13 mRNA level. Results Compared with the blank group,expressions of MMP1 /3 /13 mRNA in the control group were significantly increased（ P〈 0. 05）; compared with the control group,expressions of MMP1 /13 mRNA in the stretch group were decreased markedly（ P〈 0. 05）,while he descend range of MMP3 mRNA was not obviously（ P 〉0. 05）. Conclusion Cyclic mechanical stretch（ at a magnitude of 0-12%,frequency of 1Hz,and time of 60 min per day） may inhibit the MMP1 /13 mRNA expression in synovioblasts of human knee joints,which may have the potential to protect cartilage damage induced by synovial inflammation.

关 键 词：滑膜成纤维细胞 细胞培养 周期性牵张力 基质金属蛋白酶 

分 类 号：R274[医药卫生—中医骨伤科学]