大米α-球蛋白肽YGEGSSEEG对TNF-α诱导的血管内皮细胞损伤的影响  

Effects of rice peptide YGEGSSEEG on improving endothelial cell injury induced by tumor necrosis factor

作　　者：鞠志远 王丽丽[1] 刘丽娅[1] 周闲容[1] 周素梅[1] 佟立涛[1] JU Zhi-yuan;WANG Li-li;LIU Li-ya;ZHOU Xian-rong;ZHOU Su-mei;TONG Li-tao(Institute of Agro-Products Processing Science and Technology,Chinese Academy of Agricultural Sciences; Key Lab of Agro-Products Processing,Ministry of Agriculture,Beijing　100193,China)

机构地区：[1]中国农业科学院农产品加工研究所,农业部农产品加工综合性重点实验室,北京100193

出　　处：《中国药理学通报》2018年第4期501-507,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No31401505)

摘　　要：目的研究从大米α-球蛋白中获取的活性肽发挥抗动脉粥样硬化功效的机制。方法大米α-球蛋白经体外消化产物,采用外翻肠囊法收集可吸收组分,利用凝胶色谱和半制备高效液相色谱分离纯化,液相质谱串联测定氨基酸序列。利用TNF-α诱导人脐静脉内皮细胞损伤模型来评价肽的功效。检测细胞存活率,筛选肽的合适剂量;流式细胞仪检测细胞凋亡率;蛋白质印迹检测Bcl-2、Bax、p-p38、血管细胞黏附分子及NF-κB信号通路等蛋白表达水平;酶联免疫检测谷胱甘肽过氧化物酶等氧化应激因子含量。结果从可吸收组分中获得肽YGEGSSEEG。50 mg·L^(-1)合成肽能够明显抑制TNF-α引起的细胞凋亡,调控Bcl-2/Bax蛋白表达以及阻碍p38蛋白的磷酸化;同时,明显抑制HUVEC产生黏附分子、阻碍NF-κB信号通路的激活;此外,肽对细胞中的氧化应激水平并无明显影响。结论大米α-球蛋白肽YGEGSSEEG可通过抑制细胞凋亡和降低黏附分子表达,发挥血管内皮细胞保护作用。Aim To investigate the peptides and its protection for vascular endothelial cells,derived from the absorbed components of riceα-globulin,which was shown to be effective in anti-atherosclerosis.Methods The amino acid sequence was purified by gel chromatography and RP-HPLC,and determined by ESI/MS.Then the peptide was chemically synthesized.Human umbilical vein endothelial cell injury model was induced by tumor necrosis factor-α.The cell viability was measured by cell counting kit to screen the appropriate peptide intervention concentration.The apoptotic rate was detected by flow cytometry.Bcl-2,Bax,p-p38,vascular cell adhesion molecule and the protein expression level of NF-κB signaling pathway were detected by Western blot and immunofluorescent staining.Results Apoptosis of HUVECs induced by TNF-αwas significantly increased by YGEGSSEEG,which also regulated expression of Bcl-2/Bax proteins and inhibited phosphorylation of p38 protein.Besides,the peptide suppressed the production of VCAM-1,ICAM-1 and activation of NF-κB pathway.While it did not significantly improve the oxidative stress response in HUVECs.Conclusion Peptide YGEGSSEEG protects vascular endothelial cells through suppressing apoptosis and expression of adhesion molecules.

关 键 词：大米α-球蛋白肽 人脐静脉内皮细胞 细胞凋亡 黏附分子 NF-κB信号通路 BCL-2/BAX蛋白 

分 类 号：R322.123[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]