菜籽饼粕中多肽的提取纯化及抗肿瘤活性

Extraction,Purification and Anti-Tumor Activity of A Novel Peptide from Rapeseed Meal

从菜籽饼粕中分离纯化多肽,并验证其抗肿瘤活性。采用超声-微波协同萃取法获得菜籽饼粕总蛋白,通过碱性蛋白酶水解提取菜籽饼粕总多肽,连用分子排阻色谱和反相高效液相色谱纯化获得菜籽饼粕多肽(RMP)。通过对Hela细胞生长的抑制作用评价RMP的抗肿瘤活性。结果显示:RMP由8个氨基酸组成,序列为Asp-Val-Phe-Val-Pro-Pro-Val-Leu,相对分子质量为886 u。RMP的处理会改变Hela细胞细胞核形态,诱导细胞凋亡。作用机制涉及降低Hela细胞线粒体膜电位,促进细胞色素C从线粒体中释放,进而增强Caspase蛋白的活性,从而实现线粒体途径的细胞凋亡。因此,RMP可作为一个潜在的抗肿瘤活性物质在预防和治疗肿瘤的保健品和药品中开发利用。

In the present study,purified polypeptide was separated from rapeseed meal,and its anti-tumor activity was also verified.The total protein of rapeseed meal was obtained by ultrasonic-microwave synergistic extraction method and the total peptides were got by alkaline protease hydrolysis.RMP were separated by molecular exclusion chromatography from the total peptides and purified by reversed-phase high performance liquid chromatography(RP-HPLC).Structural analysis showed that the molecular mass of RMP was 886 u,and it had a simple amino acid sequence:Asp-Val-Phe-Val-Pro-Pro-Val-Leu.To explore the antitumor activity of RMP,the effect of RMP on Hela cell apoptosis and the underlying preventative mechanisms were investigated.Results showed that RMP reduced the viability of Hela cells in a concentration-dependent manner.TUNEL analysis and flow cytometric examination indicated that RMP decreased cell proliferation via a mitochondrial-dependent pathway,as promotion of the movement of cytochrome C from the mitochondria to the cytoplasm,and triggering of Caspase-9 and-3.Therefore,RMP might be a promising tumor preventive agent,and could be used in health care products and medicines for treatment of tumors.