电刺激对过氧化氢诱导衰老肌管细胞能量代谢的影响

Effect of Electrical Stimulation on Energy Metabolism of Senescent Myoduct Cells Induced by Hydrogen Peroxide

目的:运用电刺激肌管细胞收缩模拟骨骼肌收缩,并观察其对过氧化氢(H_(2)O_(2))诱导的C2C12细胞衰老模型能量代谢相关指标表达的影响,以此研究体外肌管收缩对衰老骨骼肌细胞能量代谢影响的分子机制。方法:使用不同浓度的H_(2)O_(2)诱导肌管细胞建立衰老模型,并用β-半乳糖苷酶染色检测衰老水平;电刺激干预培育成功的衰老肌细胞,并分为对照组(C组)、H_(2)O_(2)诱导衰老组(H组)、对照+电刺激组(CE组)、H_(2)O_(2)诱导衰老+电刺激组(HE组)。各组细胞通过Western blot法对PGC-1α蛋白表达进行检测,ATP水平测试盒测量ATP水平。结果:经过浓度为100μmol/L H_(2)O_(2)诱导肌管细胞24 h后成功构建骨骼肌细胞衰老模型。与C组比较,H组的ATP水平表达呈非常显著性的下降(P<0.01);CE组的ATP水平较C组呈非常显著性升高(P<0.01);HE组的ATP水平虽低于C组,但高于H组,且呈非常显著性差异(P<0.01);与C组相比,H组PGC-1α蛋白表达下降,呈非常显著性差异(P<0.01);HE组PGC-1α蛋白表达虽低于C组,但较H组呈非常显著性升高(P<0.01)。结论:H_(2)O_(2)是诱导骨骼肌肌管细胞衰老的一种有效方式,衰老肌管细胞的ATP和PGC-1α蛋白表达水平显著下降,进行电刺激干预后可以有效提高衰老肌管细胞ATP水平和PGC-1α蛋白表达。

Objective:Muscle tube cell contraction was simulated by electrical stimulation,and its effect on the expression of energy metabolism related indicators in C2C12 cell aging model induced by hydrogen peroxide(H_(2)O_(2))was observed,so as to study the molecular mechanism of the effect of muscle tube contraction on energy metabolism of aging skeletal muscle cells in vitro.Methods:The senescence model was established by inducing myoduct cells with different concentrations of H_(2)O_(2),and the senescence level was detected by SA-β-galactosidase staining.Electrical stimulation interfered with the successful growth of senescent muscle cells which were divided into control group(group C),H_(2)O_(2)-induced group(group H),control+electrical stimulation group(group CE),and H_(2)O_(2)-induced+electrical stimulation group(group HE).For cells in each group,PGC-1αprotein expression was measured by Western blot and ATP level was measured by ATP level test kit.Results:Myotube cells were treated with 100μmol/L H_(2)O_(2) for 24 hours,and the aging model of skeletal muscle cells was successfully established.Compared with group C,ATP level in group H was significantly decreased(P<0.01);The ATP level in CE group was significantly higher than that in C group(P<0.01).The ATP level in HE group was lower than that in C group,but higher than that in H group,and the difference was very significant(P<0.01).Compared with group C,the expression of PGC-1αin group H was significantly decreased(P<0.01).Although the expression of PGC-1αin HE group was lower than that in C group,it was significantly higher than that in H group(P<0.01).Conclusion:H_(2)O_(2) is an effective way to induce senescence of skeletal muscle myoduct cells.The protein expression levels of ATP and PGC-1αin senescent C_(2)C_(12) cells decreased significantly.After electrical stimulation intervention,ATP level and PGC-1αprotein expression in senescent C_(2)C_(12) cells could be effectively increased.