论文部分内容阅读
目的探讨不同应力作用下成肌细胞内ROS产生的量J、NK和NFkappaB的活化程度以及它们间的交互对话。方法利用Flexercell细胞加力仪对体外培养C2C12成肌细胞施加不同频率、不同幅度的牵张应力。细胞计数、MTT实验检测应力条件下细胞的活性,通过荧光染料检测ROS的生成,荧光报告系统检测NFkappaB的活性以及Western Blot检测磷酸化和总体JNK1的水平,分析三者之间的交互对话关系。结果随着周期性应力的增加,细胞内ROS生成逐渐增加。随着应力的增大,细胞的凋亡逐渐增加,与此同时J,NK1活性逐渐增高,而JNK1表达水平并没有明显改变。与ROS生成量和JNK1活化逐渐增加不同,应力在10%以下时NFkappaB的活性随着应力的增大而增大,而当应力大于15%时,NFkappaB活性开始下降。结论当ROS的量积累到一定的时候J,NK开始激活,而ROS的量较低的时候,NFkappaB被激活。
Objective To investigate the activation of intracellular ROS (ROS), NK and NFkappaB in myoblasts under different stresses and the interactive dialogue among them. Methods The strain of C2C12 myoblasts cultured in vitro were exposed to different frequencies and different amplitudes with Flexercell cell force meter. MTT assay was used to detect the activity of cells under stress, the ROS generation was detected by fluorescent dye, the activity of NFkappaB was detected by fluorescence reporter system and the level of phosphorylation and total JNK1 was detected by Western Blot. Results With the increase of periodic stress, the intracellular ROS production gradually increased. With the increase of stress, cell apoptosis gradually increased, while J, NK1 activity gradually increased, while the expression level of JNK1 did not change significantly. In contrast to the increase in ROS production and JNK1 activation, NFkappaB activity increases with increasing stress below 10% stress, whereas NFkappaB activity begins to decrease at more than 15% stress. Conclusion When NK and NK begin to accumulate when the amount of ROS accumulates to a certain level, NFkappaB is activated when the amount of ROS is low.