乳源酪蛋白糖巨肽作用人结肠癌细胞HT-29调控炎症活力研究  

Milk-Derived Casein Glycomacropeptide(CGMP) Regulates Inflammation in Human Colon Carcinoma Cells HT-29

作　　者：王泳[1] 龚建苗[1] 贾彦[1] 阎亚丽[1] 庞广昌[1] 赵培[1] 陈庆森[1] WANG Yong;GONG Jianmiao;JIA Yan;YAN Yali;PANG Guangchang;ZHAO Pei;CHEN Qingsen(Tianjin Key Laboratory of Food Biotechnology, College of Biotechnology and Food Science,Tianjin University of Commerce, Tianjin 300134, China)

机构地区：[1]天津商业大学生物技术与食品科学学院,天津市食品生物技术重点实验室,天津300134

出　　处：《食品科学》2017年第15期201-207,共7页Food Science

基　　金：国家自然科学基金面上项目(31071522);天津商业大学国家基金培育项目(160120)

摘　　要：为进一步阐释乳源酪蛋白糖巨肽(casein glycomacropeptide,CGMP)的抗炎及修复肠道炎症的功效,本研究在确定了乳源CGMP对脂多糖(lipopolysaccharides,LPS)诱导的结肠癌细胞HT-29NF-κB亚单位p65蛋白的影响和乳源CGMP调控核转录因子(nuclear factor,NF)-κB信号通路的基础上,通过构建表达乳源CGMP和NF-κB必需调节蛋白(NF-κB essential modulator,NEMO)的质粒,共转HT-29细胞,免疫共沉淀测定两个蛋白的相互作用。最后采用线粒体膜电位法测定乳源CGMP对HT-29细胞的促凋亡作用。结果表明,乳源CGMP与NEMO(NF-κB essential modulator)30 h共转HT-29细胞中,乳源CGMP与NEMO蛋白均表达,且表现出二者的相互作用。乳源CGMP可与NEMO相互作用影响NF-κB信号通路;同时乳源CGMP还促进HT-29细胞凋亡,呈时间依赖性,其中0.1μg/m L的CGMP作用效果最好。乳源CGMP与NEMO相互作用是间接作用于IκB激酶(IκB kinase,IKK)而作用于?IκB,进而影响NF-κB信号通路。研究揭示了乳源CGMP调控NF-κB信号通路的另一种途径,也充分说明乳源CGMP可通过作用多种抗炎途径发挥其调控作用。To further elucidate the anti-inflammatory activity of casein glycomacropeptide(CGMP)and its efficacy in enhancing recovery from intestinal inflammation,the effect of CGMP on the NF-κB p65subunit in colon carcinoma cells HT-29treated with lipopolysaccharide(LPS)and its role in regulating the NF-κB signaling pathway were determined.Furthermore,plasmids were constructed to express CGMP and NF-κB essential modulator(NEMO)and then used to co-transfect HT-29cells.Co-immunoprecipitation was used to identify the interaction between the expressed proteins.Finally,we detected the pro-apoptotic effect of CGMP on HT-29cells by measuring the change in mitochondrial transmembrane potential.The experimental results indicated that after30h transfection of HT-29cells,the co-expression and interaction of CGMP and NEMO were confirmed.CGMP affected the NF-κB signaling pathway through its interaction with NEMO with a concomitant increase in apoptosis of HT-29cells in a time-dependent manner.This effect of CGMP was maximum at0.1μg/mL.In conclusion,the interaction of CGMP and NEMO can indirectly affect IKK(IκB kinase)and therefore the NF-κB signaling pathway through IκB.The study revealed another way CGMP could regulate the NF-κB signaling pathway,and fully indicated that there are multiple ways CGMP could execute anti-inflammatory effect.

关 键 词：酪蛋白糖巨肽 结肠癌细胞HT-29 NF-κB信号途径 NF-κB必需调节蛋白 细胞凋亡 

分 类 号：TS252[轻工技术与工程—农产品加工及贮藏工程]