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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 effect of periodic mechanical stretching on the proliferation of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs). Methods The experimental groups were loaded with RA-FLSs for 2, 6, and 12 h at a cyclic mechanical stretching frequency of 1.0 Hz and tensile amplitudes of 3%, 6% and 9% respectively. The control group cells did not undergo stretching stimulation while maintaining the same conditions as the experimental group. After mechanical stretching, cell proliferation and activity were examined by flow cytometry and MTS. The changes of cell cycle regulatory factors (CyclinD1, CyclinE1, CDK2, P27) at gene level were detected by RT-PCR. RESULTS: After prolonged 6% and 9% stimulation for 6 and 12 h, the proliferation and activity of RA-FLSs significantly decreased (P <0.05), while the mRNA expressions of CDK2 and CyclinE1 decreased and the expression of P27 mRNA increased (P <0.05) , Cyclical mechanical stretching on CyclinD1 expression is relatively small. Conclusions The effect of cyclic mechanical stretching on the proliferation of RA-FLSs is related to the tensile strength and duration, and 6% and 9% mechanical stretch can inhibit the proliferation of RA-FLSs, and the inhibition may be Through the regulation of CyclinE1, CDK2 and P27 expression to achieve. This study is of great significance to explore the role of mechanical stimulation in the pathogenesis of rheumatoid arthritis and clinical prevention and treatment.