鹿茸水提取物及其蛋白酶解物对小鼠脾淋巴细胞增殖的影响

Effect of Aqueous Extract of Velvet Antler and Its Crude Protein Hydrolysates on Murine Splenocytes Proliferation

研究了鹿茸水提取物中非蛋白成分、粗蛋白酶解物及分离组分对小鼠脾淋巴细胞增殖的影响.非蛋白成分和酶解物的分离分别采用HPLC法和超滤法.应用WST-8法检测各分离组分对小鼠脾淋巴细胞增殖的影响,采用高分辨质谱对非蛋白成分进行初步鉴定.结果显示,非蛋白成分对未经刀豆蛋白A(ConA)诱导的小鼠脾细胞增殖的促进作用优于其分离组分,Fr.Ⅲ和Ⅳ对ConA诱导的脾淋巴细胞增殖有抑制作用,初步鉴定其为碱基和核苷酸的混合物.分子量大于10 kDa的肽段,单独可刺激小鼠脾细胞增殖,且对ConA诱导的T淋巴细胞增殖有抑制作用.肽段分子量大小对其免疫调节活性影响较大,但并未呈现直接的相关性.肽的氨基酸组成、序列及构型等也是影响其活性的重要因素.

The aim of the present study was to investigate the effect of non-protein components （ AEVA- S） , crude protein hydrolysates and their fractions from aqueous extract from velvet antler （AEVA） on routine splenocytes proliferation. AEVA-S and crude protein hydrolysates were fractionatcd by HPLC and uhrafihration, respectively. The effect of each fraction on the proliferation of murine splenocytes was de- tected by WST-8. The non-protein components were preliminary identified by high resolution mass spec- trometry. The results suggested that the stimulated effect of AEVA-S on the proliferation of resting murine splenocytes was more effective than its fractions. Fr. III and IV inhibited ConA-stimulated splenocyte proliferation significantly, and they were preliminary identified as a mixture of bases and nucleotides. Peptide fractions of MW 〉 10 kDa stimulated the proliferation of resting murine splenocytes and inhibited ConA-stimulated splenocyte proliferation. MW of peptides showed a greater impact on its immunomodulat- ing activity, but did not show a direct correlation. Amino acid composition, sequence and structure of peptides were also important factors affecting its activity.