采用醋酸溶液和乙醇提取了6支鲜二杠马鹿茸的主干顶部(未骨化部分)、中部(部分骨化)、根部(大部分骨化)3个部位的多肽,进行了多肽的SDS-PAGE分析和胰岛素样生长因子-1(IGF-1)、表皮生长因子(EGF)、神经生长因子(NGF)的Western-blotting分析.采用考马斯亮蓝G-250法进行了马鹿茸不同部位总多肽定量,RIA法测定了IGF-1含量,ELISA法比较了不同部位EGF、NGF的OD值.结果:SDS-PAGE表明鹿茸酸醇提取多肽为一系列分子量低于17 kD的混合物,Western-blotting证明其含有IGF-1、EGF、NGF等生长因子.定量分析表明:马鹿茸顶部、中部、根部总多肽、IGF-1及醇提多肽中IGF-1含量均差异显著(P<0.05),EGF、NGF ELISA检测OD值差异显著(P<0.05),均从顶部到根部递减.
We extracted the active components of different segments (top, middle & basis) of six fresh velvet antlers-polypeptides with acetic acid solution(pH 3.5)and alcohol and analyzed by SDS-PAGE and Western blotting with anti-human IGF- 1, EGF, NGF as the first antibodies. The total polypeptides and IGF - 1 contents in different segments of velvet antler were detected with Coomassie brilliant blue G - 250 and RIA techniques. EGF and NGF OD values were compared with ELISA. Results: The results of SDS -PAGE indicated that deer velvet antler polypeptides were a mix whose molecular were lower than 17 kD, and Western blotting revealed that IGF - 1, EGF and NGF were extracted with acid and alcohol. Quantitative analysis showed that polypeptides and IGF- 1 extracted with acid and alcohol, and IGF- 1 extracted with acid in different segments of velvet antler were different significantly( P 〈 0.05). The OD value difference of EGF and NGF contents between different segments of velvet antler showed by ELISA is also significant(P 〈 0.05), all the data revealed that they are highly concentrated in the top of velvet antler( P 〈 0.05). This test offered an internal quality index in subsection utilization of Cervus elaphus linnaeus.