牡蛎肽对RAW264.7巨噬细胞的免疫调节作用  

The immunomodulatory effects of oyster peptide on mouse RAW264.7 cells

作　　者：李锦弘 郑慧珍 陈慧[1,2,3,4] 刘书来 顾赛麒[1,2,3,4] 王芮 相兴伟[1,2,3,4] LI Jinhong;ZHENG Huizhen;CHEN Hui;LIU Shulai;GU Saiqi;WANG Rui;XIANG Xingwei(College of Food Science and Technology,Zhejiang University of Technology,Hangzhou 310014,China;Key Laboratory of Marine Fishery Resources Exploitment&Utilization of Zhejiang Province,Hangzhou 310014,China;National R&D Branch Center for Pelagic Aquatic Products Processing,Hangzhou 310014,China;Collaborative Innovation Center of Seafood Deep Processing,Dalian Polytechnic University,Dalian 116034,China)

机构地区：[1]浙江工业大学食品科学与工程学院,浙江杭州310014 [2]浙江省深蓝渔业资源高效开发利用重点实验室,浙江杭州310014 [3]国家远洋水产品加工技术研发分中心,浙江杭州310014 [4]海洋食品精深加工关键技术省部共建协同创新中心,大连工业大学,辽宁大连116034

出　　处：《食品与发酵工业》2023年第22期49-56,共8页Food and Fermentation Industries

基　　金：国家重点研发计划项目(2020YFD0900902);国家自然科学基金项目(32101947)。

摘　　要：为研究牡蛎肽对小鼠巨噬细胞RAW264.7免疫功能的影响及其作用机制,该研究采用细胞计数试剂盒-8分别考察了不同浓度牡蛎肽对巨噬细胞RAW264.7增殖能力的影响。通过中性红法、硝酸还原酶法、酶联免疫吸附法及实时荧光定量PCR(quantitative real-time PCR,RT-PCR)法检测巨噬细胞的吞噬能力、一氧化氮(NO)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)及白细胞介素-10(interleukin-10,IL-10)的分泌量及其mRNA表达量,然后通过蛋白免疫印迹(Western-blot)法测定巨噬细胞中核因子-κB(nuclear factor kappa-B,NF-κB)通路主要蛋白p65和IκBα的磷酸化表达情况。结果表明,质量浓度在1000μg/mL以内时,牡蛎肽对巨噬细胞增殖率无显著影响(P>0.05)。此外,牡蛎肽可增强巨噬细胞吞噬活性,显著提高RAW264.7细胞NO、TNF-α、IL-1β、IL-6及IL-10的分泌量及相关基因mRNA表达水平(P<0.05),其作用效果呈现剂量依赖性。Western-blot结果显示,与脂多糖造模组比较,牡蛎肽能显著降低RAW264.7细胞IκBα及p65蛋白磷酸化水平(P<0.05)。牡蛎肽对小鼠RAW264.7巨噬细胞具有一定的免疫调节作用,其作用机理可能和NF-κB通路的激活有关。To study the effect of oyster peptides on the immune function of mouse macrophage RAW264.7 and its mechanism,the effects of different concentrations of oyster peptides on the proliferation of RAW264.7 macrophages were investigated by cell counting kit-8 in this study.The phagocytosis of macrophages,secretion of NO,TNF-α,IL-1β,IL-6,and IL-10 and the relative mRNA expression levels of these cytokines were detected by neutral red method,nitrate reductase method,enzyme-linked immunosorbent assay,and real-time fluorescence quantitative PCR.The expression of protein p-p65/p65 and p-IκBα/IκBα in the NF-κB pathway of macrophages was detected by Western-blot.Results showed that OP concentration within 1 000 μg/mL had no significant effect on cell proliferation(P>0.05).OP could also enhance the phagocytosis of macrophages,and significantly increase the secretion of NO,TNF-α,IL-1β,IL-6 and IL-10 and the relative mRNA expression levels in RAW264.7 cells(P<0.05),while the regulating effect was dose-dependent.Western-blot results showed that OP significantly decreased the levels of IκBα and p65 phosphorylation in RAW264.7 cells compared with the lipopolysaccharide-induced group(P<0.05).These results suggested that OP had an immunomodulatory effect on mouse macrophage RAW264.7,and the mechanism may be related to the activation of the NF-κB pathway.

关 键 词：牡蛎肽 RAW264.7细胞 免疫抑制 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]