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为研究TNF α对离体孵育骨骼肌蛋白代谢及泛素系统基因表达的影响 ,采用含重组大鼠TNF α(6 0 0 0U/ml)的孵育液有氧孵育大鼠伸趾长肌 (EDL) ,对照组孵育液除不含TNF α外其余成分与实验组相同 ,高效液相色谱法测定总蛋白和肌纤维蛋白的代谢率 ,核糖核酸印迹法检测EDL泛素mRNA和C2 mRNA的表达变化。结果显示 ,实验组大鼠EDL总蛋白和肌纤维蛋白分解代谢率与对照组相比差异无统计学意义 ,但EDL泛素mRNA(2 4kb)和C2 亚基mRNA的表达较对照组分别升高约 15 1%和 5 6 % ,差异有统计学意义。提示TNF α能与骨骼肌细胞直接发生作用 ,增强细胞内泛素蛋白酶体途径的活性 ,但TNF α能否直接增强骨骼肌蛋白分解代谢尚有待进一步探讨。
In order to study the effect of TNFα on skeletal muscle protein metabolism and gene expression of ubiquitin in vitro, recombinant rat TNFα (6000U / ml) was used to incubate the rat extensor digitorum longus (EDL) ). The control group was incubated with the same components as the experimental group without TNFα. The metabolic rate of total protein and myofibrillar protein was determined by high performance liquid chromatography (HPLC), and the changes of EDL ubiquitin mRNA and C2 mRNA were detected by Western blotting. The results showed that there was no significant difference in EDL total protein and myofibrillar protein catabolism between the experimental group and the control group, but the expression of EDL ubiquitin mRNA (24 kb) and C2 subunit mRNA increased by about 15 1% and 56%, the difference was statistically significant. These results suggest that TNFα can directly interact with skeletal muscle cells and enhance the activity of ubiquitin proteasome pathway in cells. However, whether TNFα can directly enhance the catabolism of skeletal muscle remains to be further explored.