骨骼肌细胞氧化应激损伤与黄芪多糖的干预研究
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摘要
目的:建立氧化应激骨骼肌细胞模型,观察黄芪多糖(APS)对其的影响,为今后的实验提供依据。方法:过氧化氢(H_2O_2)分100μM、200μM、300μM、400μM 4组浓度干预骨骼肌细胞,每组6个复孔,分别培养12 h和24 h,MTT法检测增殖率;用100μM H_2O_2制作氧化应激骨骼肌细胞模型(模型组),给予0.2 mg/mL APS干预(干预组),并设正常组,每组6个复孔,培养24 h,MTT法检测增殖率,流式细胞仪检测凋亡率、活性氧簇(ROS)水平。结果:与正常组比较,100μM以上浓度的H_2O_2均有统计学意义(P<0.01),随着H_2O_2浓度的增加,骨骼肌细胞的增殖率显著下降(P<0.01)。与正常组比较,模型组的ROS水平及凋亡率均升高(P<0.01);干预组的ROS水平及凋亡率均下降(P<0.01),与模型组比较差异有统计学意义(P<0.01)。结论:100μM H_2O_2是氧化应激骨骼肌细胞模型的有效浓度。APS具有良好的抗氧化性,具有改善肌肉减少症的作用。
Objective: To establish the model of oxidative stress of skeletal muscle cells, and to observe the effect of astragalus polysaccharides(APS) on it, thus to provide a basis for future experiments. Methods: The skeletal muscle cells were treated with hydrogen peroxide(H_2O_2) in groups of 100 μM, 200 μM, 300 μM and 400 μM. Each group was given 6 replicate wells and cultured for 12 h and 24 h respectively. MTT was used to detect the proliferation rate. The models of oxidative stress of skeletal muscle cells(the model group) was made with 100 μM H_2O_2, and was treated with 0.2 mg/mL APS(the intervention group). The normal group was set up with 6 duplicate wells in each group, cultured for 24 h. MTT was used to detect the proliferation rate. Rate flow cytometry was applied to detect the apoptotic rate and reactive oxygen species(ROS) levels. Results: The concentration of H_2O_2 above 100 μM was statistically significant compared to that in the normal group(P<0.01). As the concentration of H_2O_2 increased, the apoptotic rate in skeletal muscle cells decreased significantly(P<0.01). Compared to the normal group, the ROS level and apoptotic rate of the model group were significantly increased(P<0.01); the ROS level and apoptotic rate of the intervention group were significantly decreased(P<0.01); there was a significant difference in it when compared to the model group(P<0.01). Conclusion: 100 μM H_2O_2 is an effective concentration of oxidative stress skeletal muscle cell model. APS has better action of anti-oxidation and it can improve muscle reduction.
Objective: To establish the model of oxidative stress of skeletal muscle cells, and to observe the effect of astragalus polysaccharides(APS) on it, thus to provide a basis for future experiments. Methods: The skeletal muscle cells were treated with hydrogen peroxide(H_2O_2) in groups of 100 μM, 200 μM, 300 μM and 400 μM. Each group was given 6 replicate wells and cultured for 12 h and 24 h respectively. MTT was used to detect the proliferation rate. The models of oxidative stress of skeletal muscle cells(the model group) was made with 100 μM H_2O_2, and was treated with 0.2 mg/mL APS(the intervention group). The normal group was set up with 6 duplicate wells in each group, cultured for 24 h. MTT was used to detect the proliferation rate. Rate flow cytometry was applied to detect the apoptotic rate and reactive oxygen species(ROS) levels. Results: The concentration of H_2O_2 above 100 μM was statistically significant compared to that in the normal group(P<0.01). As the concentration of H_2O_2 increased, the apoptotic rate in skeletal muscle cells decreased significantly(P<0.01). Compared to the normal group, the ROS level and apoptotic rate of the model group were significantly increased(P<0.01); the ROS level and apoptotic rate of the intervention group were significantly decreased(P<0.01); there was a significant difference in it when compared to the model group(P<0.01). Conclusion: 100 μM H_2O_2 is an effective concentration of oxidative stress skeletal muscle cell model. APS has better action of anti-oxidation and it can improve muscle reduction.
引文
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[4] DEMONTIS F,PICCIRILLO R,GOLDBERG A L,et al.The influence of skeletal muscle on systemic aging and lifespan[J].Aging Cell,2013,12(6):943-949.
[5] KONDO H,MIURA M,ITOKAWA Y,et al.Oxidative stress in skeletal muscle atrophied by immobilization[J].Acta Physiol Scand,1991,142(4):527-528.
[6] PHAM F H,SUGDEN P H,CLERK A,et al.Regulation of protein kinase B and 4E-BP1 by oxidative stress in cardiac myocytes[J].Circ Res,2000,86(12):1252-1258.
[7] 付绍婷,邱守涛,李茜,等.不同强度耐力训练对衰老小鼠骨骼肌线粒体抗氧化及生物发生相关基因表达的影响[J].中国运动医学杂志,2014,33(11):1069-1074.
[8] 于玲,王知斌,王秋红,等.黄芪中黄酮类化合物药理作用研究进展[J].中医药信息,2018,35(2):104-108.