汉麻活性肽对RAW264.7巨噬细胞的免疫调节作用  

Immunomodulatory Effect of Bioactive Peptides from Hemp Seeds on RAW264.7 Macrophages

作　　者：葛婧仪 张正海[1] 魏连会[1] 石杰[1] 姬妍茹[1] 李柏阳 董艳[1] GE Jingyi;ZHANG Zhenghai;WEI Lianhui;SHI Jie;JI Yanru;LI Boyang;DONG Yan(Daqing Branch of Heilongjiang Academy of Sciences,Daqing 163316,Heilongjiang,China)

机构地区：[1]黑龙江省科学院大庆分院,黑龙江大庆163316

出　　处：《食品研究与开发》2025年第8期90-96,共7页Food Research and Development

基　　金：黑龙江省省属科研院所科研业务费项目(CZKYF2021-2-B015)。

摘　　要：为研究汉麻活性肽对小鼠单核巨噬细胞RAW264.7的免疫调节作用,经酶解、超滤、葡聚糖凝胶G-15分离制备汉麻活性肽,并利用液相色谱-串联三重四极杆质谱(liquid chromatography-tandem mass spectrometry,LC-MS/MS)鉴定多肽序列。利用细胞计数试剂盒-8(cell counting kit-8,CCK-8)、生物显微镜观察、中性红吞噬实验、硝酸根还原酶法、酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)和流式细胞术测定多肽对巨噬细胞的增殖、细胞形态、吞噬功能、NO释放、相关细胞因子含量和细胞周期分布的影响。结果表明,汉麻活性肽主要由DGDGSGFF、MPEDVIAN和SYNNGDQQ等共9条多肽组成,分子量在740~1 038 Da。汉麻活性肽对RAW264.7细胞的增殖和分化均具有刺激作用,在浓度为1 000μg/mL时免疫增强效果最佳,G0/G1期细胞增加56.76%,RAW264.7细胞吞噬能力增强1.33倍,NO、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)分泌能力分别提高1.84、3.94、3.19、2.52倍。由此可知,汉麻活性肽具有激活巨噬细胞、增强机体非特异性免疫的潜在作用。The immunomodulatory effect of bioactive peptides from hemp seeds on RAW264.7 macrophages was investigated.The bioactive peptides from hemp seeds were prepared by enzymatic hydrolysis,ultrafiltra⁃tion,and Sephadex G⁃15 gel filtration chromatography.Afterwards,the peptide sequences were identified by LC⁃MS/MS.The proliferation,morphology,phagocytosis,and secretion of nitric oxide(NO),interleukin(IL)⁃1β,IL⁃6,and tumor necrosis factor(TNF)⁃αin RAW264.7 macrophages treated with the bioactive peptides were determined by the cell⁃counting kit 8,microscope observation,neutral red phagocytosis assay,nitrate re⁃ductase assay,and enzyme⁃linked immunosorbent assay.At the same time,the effect of bioactive peptides from hemp seeds on the distribution of macrophages in each phase of cell cycle was studied by flow cytometry.The results showed that the bioactive peptides from hemp seeds mainly consisted of 9 peptides,including DGDGSGFF,MPEDVIAN,and SYNNGDQQ,with the molecular weights ranging from 740 to 1038 Da.The bioactive peptides promoted the proliferation and differentiation of RAW264.7 cells,demonstrating the stron⁃gest immunoenhancing effect at the concentration of 1000μg/mL.The number of cells in the G0/G1 phase in⁃creased by 56.76%,RAW264.7 cells phagocytic ability enhanced by 1.33 times,and the secretion levels of NO,IL⁃1β,IL⁃6,and TNF⁃αwere increased by 1.84,3.94,3.19,and 2.52 times,respectively.It can be con⁃cluded that the bioactive peptides from hemp seeds had the potential role of activating macrophages and en⁃hancing non⁃specific immunity.

关 键 词：汉麻籽 汉麻活性肽 RAW264.7巨噬细胞 复合酶解 免疫调节作用 

分 类 号：R285[医药卫生—中药学]