梅花鹿鹿角脱盘总肽的分离纯化及其抗肿瘤活性研究  

Study on separation,purification and antitumor activities of total peptide from Sika Antlerbase

作　　者：胡薇[1] 田玉华[1] 孟星宇[1] 李沐[1] 李婷[1] 吴炎[1] 

机构地区：[1]吉林农业大学生命科学院,吉林长春130118

出　　处：《上海中医药杂志》2013年第7期96-100,共5页Shanghai Journal of Traditional Chinese Medicine

基　　金：吉林省产业技术研究与开发项目(JF2012C002-1);吉林省长春市科技局优质梅花鹿产业化关键技术与开发重大科技专项(2008214)

摘　　要：目的分离纯化梅花鹿鹿角脱盘总肽,以小鼠乳腺癌细胞系MA782为体外研究对象,初步研究鹿角脱盘总肽的抗肿瘤活性。方法采用酸提醇沉、真空旋转蒸发及冷冻干燥提取鹿角脱盘总肽,再经Sephadex G-25凝胶过滤层析除盐,BCA法测定总肽的蛋白质浓度,SDS-PAGE电泳分析总肽中的蛋白质,MTT法和流式细胞仪检测鹿角脱盘总肽对小鼠乳腺癌细胞系MA782增殖的影响和细胞周期的变化。结果分离纯化出鹿角脱盘总肽粗提物,其蛋白质含量为91.800%;SDS-PAGE分析显示,鹿角脱盘总肽中主要含有11 kDa,18 kDa,40 kDa,64 kDa和80 kDa 5种多肽,总肽对小鼠乳腺癌细胞的增殖具有明显抑制作用,细胞周期各期细胞比例变化明显,当总肽浓度为30 mg/L时,作用72 h后细胞抑制率最高为69.200%,G0/G1期细胞百分比为81.621%,S期细胞百分比为8.983%。结论鹿角脱盘总肽对乳腺癌细胞具有抑制作用,为深入研究鹿角脱盘的抗肿瘤活性提供了实验依据。Objective To separate and purify total peptide from Sika Antlerbase, and the mice breast cancer cell line MA782 was used as research object in vitro to study the antitumor activities of Sika Antlerbase. Methods Total peptidc was extracted from Sika Antlerbase by the method of acid extraction and alcohol precipitation, rotary evaporation in vaccum and freeze-drying. Salt was eliminated by Sephadex G-25 Gel filtering chromatography. The protein concentration of total peptide was detected by the method of BCA. The composition of total peptide was analyzed by SDS-PAGE electrophoresis. The effect of Sika Antlerbase on proliferation of the mice breast cancer cell line MA782 was detected by the method of MTT and flow cytometry. Results Crude extraction of total peptide from Sika Antlerbuse was get by separation and purification, and its protein＇ s concentration was 91. 800%. SDS-PAGE analysis showed that total peptide of antler plate contained five kinds of polypeptide （llkDa, 18kDa, 40kDa, 64kDa and 80kDa）. Total peptide of antler plate had inhibitory effect on mice with breast cancer. When the concentration of the total peptide was 301xg/mL, the cell＇ s inhibition rate was 69. 200% and the cell＇ s percentage of the G0/G1 and S phases were 81. 621% and 8. 983% after total peptide treated for 72 hour. Conclusion Total peptide of antler plate has inhibitory effect on mice breast cancer cell line MA782. It provides theoretical foundation for researching the antitumor activities.

关 键 词：鹿角脱盘 总肽 分离纯化 活性研究 

分 类 号：R284.2[医药卫生—中药学] R285.5[医药卫生—中医学]