Effect Of Different Concentrations Of ALLN On Proliferation And Apoptosis Of C2C12 Myoblasts

PurposeTo explore the effect of different concentrations of ALLN on proliferation and apoptosis of C2C12 myoblasts. MethodsC2C12 cells cultured in vitro,after each group was treated for 0 to 72 h,respectively,MTT was used to detect C2C12 cell proliferation in each group.Annexin V-FITC/PI double staining was applied to detect C2C12 cell apoptosis in each group and Giemsa dying assay observes morphology changes of apoptosis in each group after treatment for 36 h. ResultsWhen the concentration of Ca2+ in the extracellular when it reaches 16 mmol/L.MTT colorimetric assay revealed the absorbance of Ca2+ group 6 were higher than that of Ca2+ group 9 but lower than that of other experimental and control group.The absorbance of Ca2+ group was significantly lower than that of the control group(P<0.005).After 6,12,24,36 hours of intervention,the absorbance in ALLN groups 1 to 7(cultured in serum-free media containing 16 mmol/L Ca2+ and ALLN at final concentrations of 3.125,6.25,12.5,25,50,100 and 200 μmol/L) were all significantly higher than that in the 16 mmol/L Ca2+(after 6 hours:0.449±0.024,0.472±0.022,0.513±0.008,0.540±0.014,0.588±0.016,0.607±0.030,0.700±0.020 vs. 0.355±0.012,all P=0.000; after 12 hours:0.407±0.007,0.414±0.006,0.434 ± 0.004,0.441 ± 0.003,0.460 ± 0.010,0.484 ± 0.006,0.525 ± 0.006 vs.0.368±0.027, all P=0.000; after 24 hours:0.436±0.005,0.431±0.015,0.441±0.006,0.459 ± 0.013,0.527 ± 0.009,0.581 ± 0.005,0.599 ± 0.011 vs.0.368 ± 0.007, all P=0.000; after 36 hours:0.464± 0.022,0.460 ± 0.018,0.461 ± 0.007,0.434 ±0.020,0.454 ± 0.028,0.479 ± 0.006,0.524 ± 0.011 vs.0.379 ± 0.011, all P=0.000),while no significant differences were observed after 48~72 hours of intervention.After treatment for 36 hours, the apoptosis rate in ALLN 10,50,100 and 200 μmol/L groups were(6.00±1.20)%,(5.02±1.13)%,(4.89±1.111)% and(2.71±1.15)%, all significantly lower than that in the Ca2+ group [(13.70±2.30)%](all P=0.000). Giemsa staining showed apoptotic morphological changes in the Ca2+ group,which was obviously alleviated in the ALLN group. ConclusionsWhen the concentration of Ca2+ in the extracellular reaches 16 mmol/L,which may be related to its inhibiting the proliferation of C2C12 cells and promoting the apoptosis of cells;ALLN can efficiently inhibit the apoptosis of C2C12 myoblasts induced by Ca2+,and promote C2C12 the proliferation of myoblasts within a certain range of treating time and dose.