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目的:探讨高强度应力作用下体外培养成肌细胞凋亡的分子机制。方法:采用Flexercell Strain Unit系统对体外培养成肌细胞施加牵张应力,通过DNA ladder实验观察细胞凋亡情况。Western Blot检测磷酸化和总体JNK1的水平,通过NFkappa B报告系统检测其活性。通过JNK1的RNAi实验观察抑制JNK1后,是否补救被抑制的NFkappa B活性。结果:10%表面拉伸幅度的牵张应力作用24 h后,细胞形态梭形稍变长,且排列方向有一致性的趋势,DNA ladder实验证实15%以上较高强度应力引起C2C12细胞凋亡,高强度应力条件下成肌细胞凋亡过程中,JNK1通路被激活,抑制JNK1的活化,补救被抑制的NFkappa B活性。结论:高强度周期性牵张应力促进体外培养成肌细胞凋亡是通过激活JNK1通路而抑制NFkappa B活性实现的。
Objective: To investigate the molecular mechanism of myoblast apoptosis induced by high-intensity stress in vitro. Methods: Stretching stress was applied to cultured myoblasts in vitro by using Flexercell Strain Unit system. Apoptosis was observed by DNA ladder. The level of phosphorylation and overall JNK1 was detected by Western Blot and the activity was tested by the NFkappa B reporter system. Whether inhibition of NFkappa B activity was inhibited after JNK1 inhibition by JNK1 RNAi assay was observed. Results: After 24 h stretch stress induced by 10% stretch stress, the morphologies of the cells became slightly longer and aligned in a consistent manner. DNA ladder demonstrated that 15% higher stress resulted in apoptosis of C2C12 cells , JNK1 pathway is activated in the process of myoblast apoptosis under high-intensity stress, inhibiting the activation of JNK1 and restoring the inhibited NFkappa B activity. CONCLUSION: High-intensity periodic distraction stress promotes myoblast apoptosis in vitro by inhibiting JNK1 pathway and inhibiting NFkappa B activity.