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目的 探讨大鼠趋化素样因子 1和 2 (rCKLF1、2 )对成肌细胞的增殖促进作用。方法 构建pcDNA3.1/rCKLF1和pcDNA3.1/rCKLF2真核表达载体 ,并瞬时转染 2 93T细胞 ,收获培养上清。分别以3 H -TdR掺入和MTT比色法 ,分析rCKLF1和rCKLF2对小鼠肌母细胞系C2C12和原代肌卫星细胞增殖的促进作用。结果 成功地构建了rCKLF1和rCKLF2的真核表达质粒 ,并在SV4 0转化的 2 93T细胞中瞬时表达rCKLF1和rCKLF2。瞬时转染上清能够使C2C12细胞的3 H -TdR的掺入值增加 2倍或 3倍。MTT比色法分析表明 ,rCKLF1和rCKLF2的转染上清 ,对Wistar大鼠乳鼠肌卫星细胞具有增殖促进作用。结论 rCKLF1和rCKLF2能够促进骨骼肌细胞的增殖 ,为进一步探讨CKLF家族分子在骨骼肌细胞增殖分化中的作用提供了实验依据
Objective To investigate the effects of rat chemokine-like factor 1 and 2 (rCKLF1,2) on the proliferation of myoblasts. Methods The eukaryotic expression vectors pcDNA3.1 / rCKLF1 and pcDNA3.1 / rCKLF2 were constructed and transiently transfected into 293T cells. The supernatants were harvested. The effects of rCKLF1 and rCKLF2 on the proliferation of mouse myoblast cell line C2C12 and primary myogenic satellite cells were analyzed by 3 H -TdR incorporation and MTT colorimetric assay, respectively. Results The eukaryotic expression plasmids rCKLF1 and rCKLF2 were successfully constructed and rCKLF1 and rCKLF2 were transiently expressed in 293T cells transformed with SV40. Transient transfection of supernatants resulted in a 2-fold or 3-fold increase in 3H-TdR incorporation in C2C12 cells. MTT colorimetric analysis showed that rCKLF1 and rCKLF2 transfection supernatant, on Wistar rat neonatal satellite muscle cells to promote proliferation. Conclusion rCKLF1 and rCKLF2 can promote the proliferation of skeletal muscle cells and provide experimental evidence for further exploring the role of CKLF family members in the proliferation and differentiation of skeletal muscle cells