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[Effect of different concentrations of vitamin C on proliferation and apoptosis of C2C12 myoblasts]
Chun Liu1, Shenghua Sun, Qiang Zhang
1Department of Respiration, Third Xiangya Hospital, Central South University, Changsha 410013, China. liuchun7322@yahoo.com.cn
Objective:
To explore the effect of different concentrations of vitamin C on proliferation and apoptosis of C(2)C(12) myoblasts.
Methods:
C(2)C(12) 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 H(2)O(2) group (DMEM-F12 medium containing 500 μmol/L H(2)O(2)) and vitamin C group 1 to 4(DMEM-F12 medium containing 500 μmol/L H(2)O(2) 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 C(2)C(12) cell proliferation in each group. Annexin V-PI double staining was applied to detect C(2)C(12) cell apoptosis in each group after treatment for 36 h.
Results:
After the cells were treated for 36 h and 72 h, the absorbance of vitamin C group 1 to 4 were higher than that of H(2)O(2) 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 C(2)C(12) cells apoptosis rate of vitamin C group 2 to 4 was lower than that of H(2)O(2) group; The C(2)C(12) cells apoptosis rate of vitamin C group 2 and 3 was higher than that of the negative control group, while the C(2)C(12) cells apoptosis rate of vitamin C group 4 was significantly lower than that of the negative control group (P=0.009).
Conclusion:
Vitamin C can efficiently inhibit the apoptosis of C(2)C(12) myoblasts induced by H(2)O(2), and after 36 h intervention, high concentration vitamin C may promote C(2)C(12) the proliferation of myoblasts.
