紫贻贝酶解多肽体外抗肿瘤活性研究  

The Anticancer Activity of Peptide from Hydrolysates of Mytilus edulis

作　　者：杨永芳[1] 丁国芳[1] 杨最素[1] 郁迪[1] 郑玉寅[1] 李荣[1] 

机构地区：[1]浙江海洋学院食品与药学学院,浙江舟山316004

出　　处：《浙江海洋学院学报（自然科学版）》2011年第2期113-118,共6页Journal of Zhejiang Ocean University(Natural Science Edition)

基　　金：浙江省科技厅重大项目(2010C13009);浙江省自然科学基金项目(Y3100129);浙江省教育厅研究生创新科研项目(YK2008072);浙江省新苗人才计划项目(2008R40G2110028)

摘　　要：确定了紫贻贝胰蛋白酶水解时的最佳条件,研究了紫贻贝酶解多肽的体外抗肿瘤活性。采用正交实验方法,以料液比、pH、加酶量、酶解温度和酶解时间为因素,以酶解液的氨基氮含量和肿瘤细胞增殖抑制率为指标进行L16(45)正交实验,MTT法检测酶解多肽对DU-145、PC-3、H1299和MGC80-3等肿瘤细胞的抑制活性;HE染色法观察酶解多肽对肿瘤细胞形态的影响;免疫组化法检测酶解多肽对细胞中Bcl-2表达的影响。结果表明:当料液比为1:4、pH为8.5、加酶量为1 000 U/g、温度为45℃、酶解时间为5 h时酶解多肽的水解度最大;当料液比为1:4、pH为8.0、加酶量为1 500 U/g、温度为40℃、酶解时间为2 h时酶解多肽的抗肿瘤活性最强,该肽可以下调肿瘤细胞中Bcl-2的表达。紫贻贝酶解多肽具有抗肿瘤活性,可能是通过诱导细胞凋亡来实现的。To find the optimal hydrolytic condition of Mytilus edulis and study the anticancer activity of hydrolytic peptide from M.edulis.M.edulis was hydrolyzed by Trypsin.Orthogonal experiments were adopted to analyze the effect of each factor in solid-liquid ratio,temperature,time,dosage of protease,and pH on degree of hydrolysis and anticancer activity.MTT method was used to study the anticancer activity of hydrolytic peptide on DU-145,PC-3,H1299 and MGC80-3 cells.HE staining was adopted to observe the cell morphology.Immunohistochemistry method was used to investigate the expression of Bcl-2.The optimum orthogonal tests conditions were as following： add 1 500 U/g enzyme in mussel protein,1/4 for solid-liquid ratio,pH8.0,at 40 ℃ was hydrolyzed 2 h.The obtained hydrolytic peptide could down-regulated the expression of Bcl-2.The hydrolytic peptide from M.edulis had anticancer activity and inducing apoptosis effect.

关 键 词：紫贻贝 酶解 多肽 抗肿瘤活性 

分 类 号：R282.77[医药卫生—中药学]