摘要
目的探讨黄芪含药血清抑制C2C12成肌细胞凋亡的作用机制。方法通过过氧化氢(H_(2)O_(2))诱导C2C12成肌细胞建立细胞凋亡模型,细胞分为对照组、模型组、黄芪含药血清低剂量组、黄芪含药血清高剂量组、空白组、HSP70组、凋亡组、H_(2)O_(2)+HSP70组。C2C12成肌细胞活力通过CCK-8检测。流式细胞术实验检测C2C12成肌细胞的凋亡。蛋白质印迹法检测B细胞淋巴瘤(Bcl)-2、Bcl-2相关蛋白X(Bax)、活化型半胱氨酸天冬氨酸蛋白酶-3(cleaved caspase-3)、热休克蛋白70(HSP70)表达。2′,7′-二氯二氢荧光素二乙酸酯染色用于测量C2C12成肌细胞中活性氧(ROS)水平。结果与模型组比较,黄芪含药血清可显著降低H_(2)O_(2)诱导的C2C12成肌细胞凋亡率及ROS水平和cleaved caspase-3、Bax表达,增加Bcl-2、HSP70蛋白表达并存在剂量依赖性。过表达HSP70后,与凋亡组组比较,H_(2)O_(2)+HSP70组C2C12成肌细胞凋亡率及ROS水平和cleaved caspase-3、Bax表达显著降低,Bcl-2表达显著升高。结论黄芪含药血清可通过上调HSP70表达抑制C2C12成肌细胞凋亡。
Objective To investigate the mechanism of Astragalus-containing serum in inhibiting apoptosis of C2C12 myoblasts.Methods The apoptosis model of C2C12 myoblasts was established by inducing hydrogen peroxide(H_(2)O_(2)).The cells were divided into control group,model group,Astragalus-containing serum low-dose group,Astragalus-containing serum high-dose group,blank group,HSP70 group,apoptosis group,and H_(2)O_(2)+HSP70 group.The viability of C2C12 myoblasts was detected by CCK-8.Flow cytometry was used to detect the apoptosis of C2C12 myoblasts.Western blot was used to detect the expressions of B-cell lymphoma(Bcl)-2,Bcl-2-associated protein X(Bax),activated caspase-3,and heat shock protein 70(HSP70).2′,7′-Dichlorodihydrofluorescein diacetate staining was used to measure the level of reactive oxygen species(ROS)in C2C12 myoblasts.Results Compared with the model group,Astragalus-containing serum could significantly reduce the apoptosis rate,ROS level,and cleaved caspase-3 and Bax expression of C2C12 myoblasts induced by H_(2)O_(2),and increase the expression of Bcl-2 and HSP70 proteins in a dose-dependent manner.After overexpression of HSP70,compared with the apoptosis group,the apoptosis rate,ROS level,and cleaved caspase-3 and Bax expression of C2C12 myoblasts in the H_(2)O_(2)+HSP70 group were significantly reduced,and the expression of Bcl-2 was significantly increased.Conclusion Astragalus-containing drug serum can inhibit apoptosis of C2C12 myoblasts by up-regulating HSP70 expression.
作者
张磊
周锋
ZHANG Lei;ZHOU Feng(Department of Clinical Laboratory,Jiangyin Hospital of Traditional Chinese Medicine/Jiangyin Hospital Affiliated to Nanjing University of Chinese Medicine,Wuxi,Jiangsu 214400,China)
出处
《国际检验医学杂志》
2025年第13期1581-1585,共5页
International Journal of Laboratory Medicine
基金
江苏省卫生健康委科研项目(Z2023088)
江阴市科技创新专项资金项目(JY0603A011014220018PB)
江阴市中青年卫生优秀人才培养项目(JYROYT202315)。
关键词
黄芪含药血清
成肌细胞
热休克蛋白70
凋亡
Astragalus-containing serum
myoblasts
heat shock protein 70
apoptosis
作者简介
张磊,男,主管技师,主要从事中医药治疗神经及肌肉损伤的研究;通信作者:周锋,E-mail:20335832@qq.com。
