摘要
采用复合酶法水解制备牡蛎活性肽,并运用Sephadex G-25柱层析法对酶解液中的肽类进行分离纯化与鉴定;以磺基罗丹明B(SRB)比色法,台盼蓝计数法,Hochest/PI双荧光染色法和DNA Ladder等检测牡蛎活性肽诱导HeLa细胞凋亡的方式,并采用流式细胞术和实时定量PCR分析牡蛎抗活性肽诱导HeLa细胞凋亡的作用机制。经分离纯化得到牡蛎活性肽(bioactive peptide of oyster,BPO),并测得其分子质量为751D;BPO能有效抑制HeLa细胞增殖,并阻止HeLa细胞G1期向S期的转换,促使凋亡基因表达上调。BPO的作用机制与诱导凋亡基因表达上调和细胞周期阻滞有关。
Flavourzyme and trypsin were used in combination for one-step hydrolysis of oyster homogenate.The resulting hydrolysate was separated by Sephadex G-25 column chromatography.As a result,a bioactive peptide was obtained.The apoptosis inducing effect of the bioactive peptide on HeLa cells was examined by sulforhodamine B colorimetric assay,trypan blue staining assay,Annexin V-FITC/PI double staining assay and DNA ladder assay.Meanwhile,the mechanism was analyzed by flow cytometry and real-time PCR.This peptide had a molecular weight of 751 D.It could effectively inhibit the proliferation of HeLa cells,arrest the cell cycle transition from G1 to S phase and induce the up-regulation of apoptotic gene expression.Its mechanism of action was probably associated with up-regulating apoptotic gene expression and arresting the cell cycle.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2012年第21期290-294,共5页
Food Science
基金
广东省教育部省部产学研合作专项(2010B090400461)
广东省科技计划项目(2010B020201015)
广东省自然科学基金研究团队项目(S2011030005257)
