血清浓度对心肌细胞自噬的影响

The effect of different serum concentrations on cardiomyocytes' autophagy

目的观察不同血清浓度对心肌细胞自噬的影响。方法以大鼠胚胎心肌细胞系H9c2为标本,建立3个时间点6、12、24h,以及四个不同浓度0、2、5、10%来检测细胞的自噬程度。运用Western Blot检测心肌细胞LC3Ⅱ、P62的变化,吖啶橙染色检测酸性膜泡。结果培养6h之前,较高浓度10%血清自噬程度最低,6~12h随着血清浓度增高自噬被促进。24h后去血清组促进自噬,添加2,5,10%血清均能抑制自噬。结论在筛选促进自噬的因子时,应选择背景自噬较低的模型,作用效果在6h之内的因子首选10%血清培养的心肌细胞,作用效果在6~12h之间的选择无血清培养体系,作用效果在24h之后的可选择有血清培养体系。

Objective To investigate the effect of different serum concentrations on cardiomyocytes＇ autophagy.Methods Rat embryonic cardiac cell line H9c2 was taken as specimens to detect the levels of cellular autophagy in three time points of 6,12,24 hand four different concentrations of 0,2,5,10%.Western Blot was used to detect the changes of cardiac cell LC3 Ⅱand P62,and acridine orange staining was used to detect acidic vesicles.Results Before 6h,10% concentrated serum had the lowest level of autophagy.During 6~12hours,autophagy was promoted with the increasing of serum concentrations.After 24 h,serum-deprived group promoted autophagy.Adding 2,5,10% serum could inhibit autophagy.Conclusion To find out the factors of promoting autophagy,the models with lower autophagy should be selected.Within 6hours,10% concentrated serum cultured cardiac cells are the first choice.During 6~12hours,serum-deprived culture system should be used.After 24 hours,serum culture system can be used.