采用MTT法测定活性肽对正常小鼠骨髓有核细胞的增殖作用,利用高效液相色谱-2,4-二硝基氟苯柱前衍生法测定4个活性肽组分的氨基酸组成。结果表明:小鼠骨髓细胞培养时间为24 h,泰和乌骨鸡活性肽质量浓度为0.1、1、10、100μg/mL,组分7、8、9、10可极显著(P〈0.01)促进小鼠骨髓有核细胞增殖,增殖率达130%以上。在低质量浓度(0.1μg/mL)条件下,组分7能显著促进小鼠骨髓有核细胞增殖(P〈0.01)且重现性较好,组分8可刺激小鼠骨髓有核细胞增殖298%。氨基酸组成分析表明,组分7、8、9、10均含有丰富的酪氨酸、甘氨酸、谷氨酸、天冬氨酸和丙氨酸,可以推测该5种氨基酸组成了刺激小鼠骨髓有核细胞增殖活性成分的物质基础。
The present study aimed at exploring the effect of Taihe Black-bone silky fowl active peptides and their 13 separated fractions on the proliferation of bone marrow nucleated cells from normal mice.MTT method was used to investigate the effect of active peptides on the proliferation of bone marrow nucleated cells from normal mice.HPLC precolumn derivation with 2,4-dinitrofluorobenzene method was used to determine the amino acid compositions of four active peptide fractions.When mice bone marrow nucleated cells were cultivated for 24 h,it was found that the fractions 7,8,9 and 10 separated from Taihe Black-bone silky fowl active peptides exhibited significantly strong activity(P0.01) in stimulating mice bone marrow nucleated cells,and the proliferation rate could be up to 130%.Especially,at lower concentration(0.1 μg/mL),fraction 7 was able to significantly stimulate mice bone marrow nucleated cells(P0.01) and revealed better reproducibility.Fraction 7 was capable of stimulating mice bone marrow nucleated cells proliferation by 298%.Amino acid composition analysis demonstrated that the main amino acids of fraction 7,8,9 and 10 consisted of tyrosine,glycine,glutamic acid,aspartate and alanine.It could be presumed that tyrosine,glycine,glutamic acid,aspartate and alanine may be the material base of active component in stimulating mice bone marrow nucleated cells.