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目的探讨机械应力加载是否干扰体外培养C2C12成肌细胞自身抗原及TLR3的转录表达。方法对生长于BioFlex柔性基底膜表面的C2C12细胞施加不同频率的拉伸加载,流式细胞术分析细胞增殖情况。Real-time RT-PCR检测并比较分析应力加载前、后骨骼肌自身抗原(HRS、Mi-2、U1-70、DNA-PKcs)、TLR3的mRNAs表达差异。结果 0.25 Hz、10%幅度、2 h/d的拉伸条件能加速C2C12细胞进入细胞周期。2 d和4 d拉伸组DPI较对照组提高(P<0.05),尤以2d拉伸组细胞增殖最为显著。拉伸2 d时,C2C12细胞内HRS、Mi-2、U1-70、DNA-PKcs以及TLR3的mRNA水平均显著低于同一培养时间的对照组(P<0.05)。结论机械应力刺激信号可能参与下调与骨骼肌组织炎症相关的免疫调节分子。
Objective To investigate whether mechanical stress loading interferes with the expression of autoantigen and TLR3 in cultured C2C12 myoblasts in vitro. Methods C2C12 cells grown on the surface of BioFlex flexible basement membrane were subjected to tensile loading at different frequencies, and cell proliferation was analyzed by flow cytometry. The mRNA expression of skeletal muscle autoantigens (HRS, Mi-2, U1-70, DNA-PKcs) and TLR3 were detected by Real-time RT-PCR and compared before and after stress loading. Results Tensile conditions of 0.25 Hz, 10% amplitude and 2 h / d accelerated C2C12 cells into the cell cycle. The DPI in the 2 d and 4 d stretch groups was higher than that in the control group (P <0.05), especially in the 2 d stretch group. The mRNA levels of HRS, Mi-2, U1-70, DNA-PKcs and TLR3 in C2C12 cells were significantly lower than those in the control group (P <0.05) at the same incubation time. Conclusion The signal of mechanical stress stimulation may be involved in the down-regulation of immunoregulatory molecules associated with skeletal muscle inflammation.