热休克蛋白70对骨骼肌细胞纤维类型分化的影响-东南大学学报医学版

热休克蛋白70对骨骼肌细胞纤维类型分化的影响

单位：1. 南京中医药大学康复医学系, 江苏南京 210001;
2. 德国ULM大学康复医学科, 德国 ULM

分类号：Q782;Q813.1

出版年·卷·期（页码）：2013·32·第六期（708-713）

摘要：

目的：探讨高表达热休克蛋白70（heat shock protein 70，Hsp70）对骨骼肌细胞（C2C12细胞）纤维类型分化的影响。方法：通过构建重组pTRE2hyg-Hsp70质粒稳定转染C2C12细胞系，建立Hsp70高表达的C2C12细胞系。经过诱导分化形成骨骼肌纤维，观察不同时间点（0、3、7 d）C2C12细胞的分化情况，并检测骨骼肌类型标志肌球蛋白重链（myosin heavy chain，MHC）的表达情况。结果：与对照组相比，高表达Hsp70的C2C12细胞系分化后的3、7 d，MHCⅠmRNA及蛋白的表达明显增高（P<0.05），而在分化后7 d，MHCⅡb mRNA及蛋白的表达明显下降（P<0.05）。结论：高表达Hsp70的骨骼肌细胞C2C12可以诱导MHCⅠ的表达，促进骨骼肌细胞向Ⅰ型肌纤维转化。

Objective: To examine the effect of over-expression heat shock protein (Hsp)70 on the skeletal muscle fiber transformation 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 expression of myosin heavy chain (MHC) in over-expression Hsp70 C2C12 cells was evaluated by real-time PCR and Western blot at different time points(0,3,7 d) after the differentiation of C2C12 cells. Results: Compared with controls,the mRNA and protein expression of MHCⅠ was significantly increased(P<0.05)at different time points(3,7 d) after differentiation,whereas the expression of MHCⅡb was decreased(P<0.05)at 7 d after differentiation. Conclusion: The C2C12 cells of over-expression Hsp70 can induce the enhancement of MHCⅠ expression and facilitate the transformation of type Ⅰ fiber from C2C12 cells.

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