Effect of different concentrations of calpain inhibitor ALLN on proliferation and apoptosis of C2C12 myoblasts
10.3760/cma.j.issn.1674-635X.2015.01.007
- VernacularTitle:不同浓度钙蛋白酶抑制剂ALLN对C2C12成肌细胞增殖和凋亡的影响
- Author:
Sizeng CHEN
;
Zhiqiang DAI
;
Chunfa WENG
;
Qinglin YE
- Publication Type:Journal Article
- Keywords:
ALLN;
C2C12 myoblasts;
Proliferation;
Apoptosis
- From:
Chinese Journal of Clinical Nutrition
2015;23(1):35-40
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of different concentrations of ALLN on proliferation and apoptosis of C2C12 myoblasts.Methods After intervention with Ca2+ and ALLN,methyl thiazolyl tetrazolium and flow cytometry were used to determine the effect of Ca2+ and ALLN on the proliferation and apoptosis of C2C12 cells,respectively.The morphological changes of C2C12 myoblasts were observed using Giemsa staining.Results The absorbance of Ca2 + group was significantly lower than that of the control group (P ＜0.05).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 3.125,6.25,12.5,25,50,100,200 μmol/L) were all significantly higher than that in the 16 mmol/L Ca2+ group (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.0.386 ± 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±1.11)％,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 were obviously alleviated in the ALLN group.Conclusions Ca2+ at a concentration of 16mmol/L can induce apoptosis of C2C12 cells.In contrast,ALLN can inhibit cell apoptosis and promote proliferation in a time-and dose-dependent manner.
