周期性机械拉伸对类风湿关节炎成纤维样滑膜细胞增殖的影响  

Effects from cyclic mechanical stretch on proliferation of rheumatoid arthritis fibroblast-like synoviocytes

作　　者：张非飞 颜艳[1,2] 吕永钢 钟莉[1,2] 宋关斌[1,2] Mohanad Khalid Ahmed[1,2] 杨力 

机构地区：[1]重庆大学"生物流变科学与技术"教育部重点实验室生物工程学院,重庆400044 [2]重庆大学生物工程学院国家"111计划"基地,重庆400044

出　　处：《医用生物力学》2012年第6期692-697,702,共7页Journal of Medical Biomechanics

基　　金：国家自然科学基金资助项目(11032012,30870608);重庆市科委科技攻关重大项目资助(2009AA5045);新世纪优秀人才支持计划资助(NCET-10-0879)

摘　　要：目的探讨周期性机械拉伸对类风湿关节炎成纤维样滑膜细胞(RA-FLSs)增殖能力的影响。方法实验组细胞在周期性机械拉伸频率为1.0 Hz、拉伸幅度为3%、6%和9%的条件下,分别对RA-FLSs加载2、6和12 h。对照组细胞在保持与实验组培养条件一致的情况下不进行拉伸刺激。机械拉伸后,用流式细胞术和MTS检测细胞的增殖和活性。RT-PCR检测加载后细胞周期调控因子(CyclinD1、CyclinE1、CDK2、P27)在基因水平上的表达变化。结果 6%和9%的拉伸刺激持续作用6、12 h使RA-FLSs增殖和活性显著降低(P<0.05),同时CDK2和CyclinE1的mRNA表达降低,P27 mRNA的表达增高(P<0.05),周期性机械拉伸对CyclinD1表达的影响相对较小。结论周期性机械拉伸对RA-FLSs增殖能力的影响与拉伸强度以及持续的时间有关,6%和9%的机械拉伸刺激可以抑制RA-FLSs的增殖,而这种增殖抑制作用可能是通过调控CyclinE1,CDK2和P27的表达来实现的。本研究对于探讨力学刺激在类风湿性关节炎的发病机制以及临床防治中的作用具有一定意义。Objective To investigate the effects from cyclic mechanical stretch on proliferation of rheumatoid ar- thritis fibroblast-like synoviocytes （RA-FLSs）. Methods In the experimental groups, cyclic mechanical stretch, with frequency of 1.0 Hz and magnitude of 3%, 6% and 9%, respectively, was applied to RA-FLSs for 2 h, 6 h, and 12 h. The control group remained in the same culture condition as the experimental groups, but without any mechanical stretch. After mechanical loading, the cell viability was analyzed by MTS, and its proliferation was as- sayed by flow cytometry. RT-PCR was used to measure the gene expression of the cell cycle regulatory factors, including CDK2, cyclinD1, cyclinE1, and P27. Results Cyclic mechanical stretch with magnitude of 6% and 9% for 6 h or 12 h significantly decreased the cell proliferation and viability in RA-FLSs （P〈0.05）. Meanwhile, both CDK2 and cyclinE1 gene expressions were significantly decreased, but P27 gene expression was increased （ P 〈 0.0.5 ）. The cyclic mechanical stretch had relatively small influence on expression of cyclinD1 （ P 〉 0.05 ）. Conclu- sions The effects from cyclic mechanical stretch on proliferation of RA-FLSs depend on the stretch magnitude and duration. Mechanical stretch with magnitude of 6% and 9% can inhibit RA-FLSs proliferation, which may be achieved by regulating the expression of Cyclin E], CDK2 and P27. This study provides references for investiga- ting the role of mechanical stimulation in pathogenesis of rheumatoid arthritis, as well as its prevention and treat- ment.

关 键 词：机械拉伸 滑膜细胞 细胞增殖 细胞表达 类风湿关节炎 

分 类 号：R318.01[医药卫生—生物医学工程]