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热休克蛋白70过表达对骨骼肌细胞内ATP水平的影响
引用本文:王磊,王尊,刘跃飞,顾一煌. 热休克蛋白70过表达对骨骼肌细胞内ATP水平的影响[J]. 中华临床医师杂志(电子版), 2013, 0(24): 225-227
作者姓名:王磊  王尊  刘跃飞  顾一煌
作者单位:[1]南京中医药大学康复医学系,210001 [2]德国ULM大学康复医学科,210001
基金项目:江苏省高校自然科学基金面上项目(107KJB320010);全国高校博士点基金项目(20123237120008);江苏省高校优势学科建设工程一期项目中医学优势学科开放课题(YS2012ZYC406)
摘    要:
目的:探讨热休克蛋白70(heat shock protein 70,Hsp70)过表达对骨骼肌细胞(C2C12)内ATP水平的影响。方法通过构建重组pTRE2hyg-Hsp70质粒,稳定转染C2C12细胞系,建立Hsp70过表达的C2C12细胞系。分别在转染后细胞培养的不同时间点(0 d,3 d,7 d),检测细胞内ATP的水平。结果 Hsp70过表达的C2C12细胞系在培养的3 d,7 d,细胞内ATP的水平分别达到(14.5±2.87)nmol/mg蛋白质、(15.3±3.12)nmol/mg 蛋白质,与对照组相比明显增高(P<0.05)。结论 Hsp70过表达的骨骼肌细胞可以提高细胞内的ATP水平,提示Hsp70对骨骼肌细胞的能量代谢产生影响。

关 键 词:HSP70热休克蛋白质类  肌纤维  腺苷三磷酸

Effect of over-expression Hsp70 on the intracellular ATP level in C2C12 cells
WANG Lei,WANG Zun,LIU Yue-fei,GU Yi-huang. Effect of over-expression Hsp70 on the intracellular ATP level in C2C12 cells[J]. Chinese Journal of Clinicians(Electronic Version), 2013, 0(24): 225-227
Authors:WANG Lei  WANG Zun  LIU Yue-fei  GU Yi-huang
Affiliation:.( Department of Rehabilitation Medicine, Nanfing University of Chinese Medicine, Nanjing 210001, China)
Abstract:
Objective To examine the effect of over-expression Hsp70 on the intracellular ATP level in C2C12 cells. Methods Hsp70 gene was amplified from pAT153 plasmids and then cloned into pTRE2hyg vector. After the transfection of recombinant plasmids of pTRE2hyg-Hsp70 into the C2C12 cells, the expression of Hsp70 was examined by Western blot. Furthermore, the intracellular ATP level was evaluated in C2C12 cells at different time points (0 d, 3 d, 7 d) during cell culture. Results Compared with controls, the intracellular ATP level was significantly increased (P〈0.05) at different time points,(14.5 ± 2.87)nmol/mg protein (3 d), (15.3 ± 3.12) nmol/mg protein(7 d) after transfection. Conclusion The C2C12 cells of over-expression Hsp70 can increase the intracellular ATP level, indicating that Hsp70 may play a role in the metabolism in skeletal muscle cells.
Keywords:HSP70 heat-shock proteins  Muscle fibers  Adenosine triphosphate
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