羊胎盘肽对L6成肌细胞增殖能力和ATP浓度的影响

Cell proliferation and ATP levels of myoblast after treated with sheep placenta peptide

关键词:  羊胎盘  分子量  L6成肌细胞  增殖  ATP

Key words:  SPP  molecule weight  L6 cells  cell proliferation  ATP

摘要:

[目的]将羊胎盘肽粗品按照不同的分子量段进行划分，计算出不同分段的分子量分布，研究不同分子量段的羊胎盘肽对大鼠L6骨骼肌成肌细胞增殖能力和三磷酸腺苷（ATP（浓度的影响。[方法]采用凝胶过滤色谱法，将羊胎盘肽粗品进行分离，划分为不同的分子量段，以噻坐蓝（MTT）比色法检测不同分段羊胎盘肽对大鼠L6骨骼肌成肌细胞增殖的影响，并且采用增强型ATP检测试剂盒检测其对L6成肌细胞的ATP含量的影响。[结果]凝胶过滤色谱法分离羊胎盘肽划分为六段，其中F5、F6已不属于多肽或者氨基酸，故去除。MTT结果显示SPP（羊胎盘肽粗品）、F1、F2、F3均对大鼠L6骨骼肌成肌细胞增殖出现促进作用，其中羊胎盘中的组分F3促增殖作用最为显著；F4则出现轻微抑制的作用。最终确定除F3选用5 μg/mL以外，其他各组均选用20 μg/mL的加药浓度进行ATP浓度的检测。经过ATP实验，SPP和F3显著性促进骨骼肌成肌细胞的ATP浓度的升高（P<0.01），并且F3比SPP效果更好。而F1、F2和F4则促进ATP产生的作用并不显著。[结论]SPP以及其中的的F1、F2、F3、F4对于L6骨骼肌成肌细胞增殖能力的影响和ATP浓度的影响较为一致。其中，F3很可能是羊胎盘肽中的活性最好的部分，即分子量在260~89Da的部分。

Abstract:

[Objective] The aim of this study was to investigate the cell proliferation and ATP levels of sheep placenta peptide(SPP) with different molecule weight on myoblast(L6 cells).[Methods] SPP were isolated with different molecule weight through gel filtration chromatography and treated to L6 cells. Cell proliferation effect of SPP on L6 cells were investigated through MTT assays. Moreover, ATP levels of L6 cells after treated with SPP were measured.[Results] SPP were isolated into 6 segments of with different molecule weight(F1 to F6). The F5 and F6 segments, which were not amino acid and polypeptide, were excluded. F1, F2 and F3 segments could significantly increase the proliferation of L6 cells whereas F4 segment showed slightly inhibition effect of proliferation on L6 cells. Moreover, F3 segment could significant increase the ATP level in L6 cells(P<0.01).[Conclusion] F3 segment of SPP, with the molecule weight of 260~89 Da, showed better biological effects on L6 cells.