黄芪多糖对骨骼肌成肌细胞增殖作用的研究

Effect of Astragalus Polysaccharides on the Proliferation of Skeletal Myoblasts

目的:观察黄芪多糖(APS)对C2C12骨骼肌成肌细胞(成肌细胞)的增殖作用。方法:将成肌细胞接种到96孔培养板,APS分为0.1、0.2、0.4、0.8和1 mg/mL 5组浓度;地塞米松(DEX)分为1、10、100和1 000μM 4组浓度,培养12 h、24 h,MTT法检测计算增殖率和抑制率。根据上述实验确定APS、DEX浓度,再分对照组、DEX组、DEX+APS组,培养24 h,MTT法检测OD值、流式细胞仪检测凋亡率、光镜检测肌管横径。结果:APS在0.6 mg/mL浓度以下对成肌细胞有增殖作用(P<0.05),并在0.2 mg/mL达到峰值。100μM浓度以上DEX对成肌细胞有抑制作用(P<0.05)。用100μM DEX建立成肌细胞萎缩模型,给予0.2 mg/mL APS干预后,细胞OD值和肌管横径均升高(P<0.05),凋亡率下降(P<0.05)。结论:APS对成肌细胞有良好的增殖作用,100μM DEX可作为成肌细胞萎缩模型造模有效浓度。

Objective:To observe the effect of astragalus polysaccharides(APS)on the proliferation of C2C12 skeletal myoblasts.Methods:The well-growth myoblasts were inoculated into 96-well culture plates,and the concentrations of APS were divided into 0.1 mg/mL,0.2 mg/mL,0.4 mg/mL,0.8 mg/mL,and 1 mg/mL;the concentrations of dexamethasone(DEX)were 1μM,10μM,100μM,and 1 000μM.The culture durations were 12 h and 24 h.MTT method was used to detect the proliferation rate and the inhibition rate.The APS and DEX concentrations were determined according to the above experiments,and were divided into the control group,the DEX group,and the DEX+APS group.After 24 hours of culture,OD value was detected by MTT,apoptosis rate was detected by flow cytometry,and transverse diameter of myotube was detected by microscopy.Results:APS has a proliferate effect on myoblasts below 0.6mg/mL(P<0.05),and the effect peaks were at 0.2 mg/mL.DEX above 100μM concentration had inhibited effect on myoblasts(P<0.05).The myocardium atrophy model was established with 100μM DEX.After intervention with 0.2 mg/mL APS,the OD value of the cells and the transverse diameter of the myotubes was increased(P<0.05),and the apoptotic rate was decreased(P<0.05).Conclusion:APS has a good proliferative effect on myoblasts,100μM DEX can be used as an effective concentration for the model of myoblast atrophy.