不同浓度的维生素C对C2C12成肌细胞增殖和凋亡的影响

目的：观察不同浓度的维生素C（VitC）对体外培养的骨骼肌成肌细胞（C2C12）增殖、凋亡的影响。方法：取体外培养的C2C12细胞，胰酶消化后制成单细胞悬液，离心重悬，调整细胞密度，按200 μL/孔接种，常规培养，待细胞融合率达80%且未出现细胞分化时，将细胞分6组：阴性对照组（C2C12 细胞中加入单纯无血清培养基）、H2O2组（C2C12 细胞中加入含500 μmol/L H2O2的无血清培养基）和VitC 1，2，3，4组（C2C12 细胞中加入含500 μmol/L H2O2的无血清培养基的基础上，分别加入不同浓度的VitC：10，20，60，100 mg/L）。分别在干预0，6，24，36，48，72 h后采用四甲基偶氮唑盐法（MTT）检测各组C2C12细胞的增殖。在干预36 h后采用Annexin V-PI双染法检测各组C2C12细胞凋亡率。结果：在干预36和72 h时，各浓度的VitC组的吸光度（OD）值均较H2O2组显著升高（P＜0.001），其中以VitC 4组升高最为显著，而且VitC 4组在干预36 h时的吸光度值高于阴性对照组（P＜0.05）；在干预36 h时，VitC 2，3和4组的C2C12成肌细胞凋亡率明显低于H2O2组；VitC 2和3组的C2C12成肌细胞凋亡率高于阴性对照组，而VitC 4组的C2C12成肌细胞凋亡率明显低于阴性对照组，差异有统计学意义（P＜0.01）。结论：VitC能够有效抑制H2O2诱导的C2C12成肌细胞的凋亡，高浓度的VitC可能在干预36 h时对C2C12成肌细胞有促增殖的作用。

Effect of different concentrations of vitamin C on proliferation and apoptosis of C2C12 myoblasts

ObjectiveTo explore the effect of different concentrations of vitamin C on proliferation and apoptosis of C2C12 myoblasts. MethodsC2C12 cells cultured in vitro were collected by trypsinization to form monoplast suspension, and then centrifuged to float again. The cellular numbers were counted and the cell suspension density adjusted, and the cells were inoculated into 96-shadow mask according to 200 μL per hollow. All cells were cultured in the normal way. While cell fusion ratio arrived 80% and cells did not differentiate, cells were divided into 6 groups: a negative control group (pure DMEM-F12 medium), an H2O2 group (DMEM-F12 medium containing 500 μmol/L H2O2) and vitamin C group 1 to 4(DMEM-F12 medium containing 500 μmol/L H2O2 and 10, 20, 60, and 100 mg/L vitamin C, respectively). After each group was treated for 0, 6, 24, 36, 48,and 72 h, respectively, MTT was used to detect C2C12 cell proliferation in each group. Annexin V-PI double staining was applied to detect C2C12 cell apoptosis in each group after treatment for 36 h. ResultsAfter the cells were treated for 36 h and 72 h, the absorbance of vitamin C group 1 to 4 were higher than that of H2O2 group (P＜0.001). The absorbance of vitamin C group 4 was the highest among all the groups, significantly higher than that of the negative control group when the cells were treated for 36 h (P＜0.05). When the cells were treated for 36 h, the C2C12 cells apoptosis rate of vitamin C group 2 to 4 was lower than that of H2O2 group; The C2C12 cells apoptosis rate of vitamin C group 2 and 3 was higher than that of the negative control group, while the C2C12 cells apoptosis rate of vitamin C group 4 was significantly lower than that of the negative control group (P=0.009). ConclusionVitamin C can efficiently inhibit the apoptosis of C2C12 myoblasts induced by H2O2, and after 36 h intervention, high concentration vitamin C may promote C2C12 the proliferation of myoblasts.