红景天苷联合姜黄素促进C2C12细胞成肌分化并防止棕榈酸诱导的肌管萎缩研究

A study of promotion of myogenic differentiation of C2C12 cells by salidroside combined with curcumin and prevention of pal-mitic acid-induced myotube atrophy

中文关键词: 肌少症性肥胖  红景天苷  姜黄素  衰老  肌萎缩

英文关键词: sarcopenic obesity  salidroside  curcumin  senescence  muscular atrophy

中文摘要:

      目的 探索红景天苷、姜黄素及二者联合在棕榈酸诱导的C2C12细胞肌管分化、肌管衰老、肌管萎缩及脂质沉积中的作用.方法 采用CCK8实验研究棕榈酸、红景天苷、姜黄素的适宜浓度;通过棕榈酸与C2C12肌管的共孵育构建肌少症性肥胖细胞模型,并用红景天苷、姜黄素及二者联合同时干预;采用苏木素-伊红染色和免疫荧光检测肌管直径、β-半乳糖苷酶染色检测细胞衰老程度、油红染色检测肌管内成脂情况;采用蛋白印迹检测MyoD1、Myogenin、TRIM63、FBX32、P21、PPAR-γ的表达水平,并通过多指标流式联合分析仪检测肌管的炎症因子水平.结果 与对照组相比,400 μM红景天苷和20 μM姜黄素单独或联合干预,能够改善棕榈酸诱导的C2CI2细胞成肌分化蛋白Myod1和Myogenin表达减少、成熟肌管直径缩短、肌萎缩蛋白TRIM63和FBX32表达增加、衰老指标β-半乳糖苷酶染色阳性面积增加、衰老相关蛋白P21表达增加,以及肌管内脂质沉积和炎症的水平升高.红景天苷联合姜黄素的干预效果优于各自的单独干预.结论 红景天苷和姜黄素可能抑制棕榈酸诱导的C2CI2肌萎缩并改善肌萎缩相关因素.

英文摘要:

      Objective To explore the roles of salidroside,curcumin,and their combination in palmitic acid-induced myotube differentiation,myotube senescence,myotube atrophy,and lipid deposition in C2C12 cells.Methods The suitable concentrations of palmitic acid,salidroside and curcumin were studied by CCK8 experiment.The sarcopenic obesity cell model was established through the co-incubation of palmitic acid and C2C12 muscle tube,and salidroside,curcumin and their combi-nation were used to intervene the cell model at the same time.Hematoxylin-eosin staining and immunofluorescence test were used to detect the diameter of myotube,β-galactosidase staining was used to detect the degree of cell senescence,and oil red staining was used to detect lipid formation in the myotube.The expression levels of MyoD1,Myogenin,TRIM63,FBX32,P21 and PPAR-γ were detected by Western blotting.The levels of inflammatory factors in the myotube were detected by multi-index flow analyzer.Results Compared with the control group,the intervention of 400 μM salidroside and 20 μM curcumin alone or in combination could improve the reduction of expression of myogenic differentiation proteins Myod1 and Myogenin in-duced by palmitic acid in C2CI2 cells,shortening of the diameter of mature myotubes,increased expression of muscle atrophy-related proteins TRIM63 and FBX32,increased positive area of senescence indicator β-galactosidase staining,increased expres-sion of senescence-related protein P21,and elevated levels of lipid deposition and inflammation in the myotubes.The interven-tion effect of salidroside combined with curcumin was better than that of each intervention alone.Conclusion Salidroside and curcumin may inhibit palmitic-induced C2CI2 muscular atrophy and improve the factors associated with muscular atrophy.