小鼠淋巴细胞内多肽的分离鉴定

Isolation and Identification of Peptides in Mouse Lymphocytes

本课题以小鼠脾淋巴细胞为研究对象,对受到刺激前后细胞内多肽进行分离和鉴定。静息状态下淋巴细胞作为对照组,实验组用LPS刺激,均采用RPMI-1640不完全培养液养,破碎,收集胞内多肽,采用纳升液相-QExactive四极杆超高分辨轨道阱质谱仪(NanoLC-QExactivePlus)进行分析和鉴定,再通过网站搜索比对氨基酸序列确定是否为新发现多肽。结果表明,从静息状态下和受到刺激后小鼠脾淋巴细胞中共分离鉴定出385种多肽,肽链长度为8~30个氨基酸,其中139种为新发现多肽。

In this study,mouse spleen lymphocytes were used as the research object to isolate and identify the intracellular polypeptides before and after stimulation.The lymphocytes in the resting state were served as the control group,and the experimental group were stimulated with LPS.All of them were cultured in RPMI-1640 incomplete culture medium,then broken,collected intracellular peptides,which were identified and analyzed by nanoliter liquid phase-Q Exactive quadrupole ultra-high resolution orbitrap mass spectrometer(Nano LC-Q Exactive Plus).Finally,the amino acid sequences were searched and compared through the websites to determine whether they were newly discovered.The results showed that 385 polypeptides were isolated and identified from mouse splenic lymphocytes in resting state and after stimulation,of which 139 were newly discovered peptides.The length of peptide chains were 8-30 amino acids.This study revealed the variety and quantity of bioactive peptides in spleen lymphocytes of mice before and after being stimulated by adverse external stimuli,laying a scientific foundation for the development of bioactive peptide health products and special medical formula foods.