摘要
为探讨高核苷酸酵母水解物对RAW264.7小鼠巨噬细胞免疫活性调节及其相关作用机制,采用脂多糖(1μg/mL)刺激RAW264.7细胞来建立体外细胞炎症模型,以不同质量浓度的高核苷酸酵母水解物干预来明确其抗炎效果。ELISA法检测细胞培养上清中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)的含量,RT-PCR法检测相关mRNA表达水平,Western blot法检测细胞iNOS及胞核内核转录因子NF-κBp65蛋白水平。结果表明:高核苷酸酵母水解物作用RAW264.7细胞的安全范围≤150μg/mL。与LPS模型组相比,质量浓度60～150μg/mL的高核苷酸酵母水解物能显著增强RAW264.7吞噬能力,高核苷酸酵母水解物(60～150μg/mL)能有效抑制脂多糖(lipopolysaccharide,LPS)诱导RAW264.7细胞释放NO、TNF-α、IL-1β和IL-6炎症因子及相关mRNA表达,降低iNOS及NF-κB蛋白水平。研究结果证实了高核苷酸酵母水解物对LPS刺激RAW264.7细胞炎症的保护作用,作用机制与NF-κB通路有关,为食疗干预慢性病提供一定的理论依据。
This study aimed to investigate the effects of high-nucleotide yeast hydrolysates on the immune activities of macrophages in RAW264.7 cells and the related mechanisms. In vitro cell inflammation model was created by stimulating RAW264.7 cells with 1 μg/mL lipopolysaccharide(LPS). Different concentrations of high-nucleotide yeast hydrolysates were used as interventions to determine their anti-inflammatory effects. ELISA was used to detect the contents of interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), and interleukin-1β(IL-1β) in cell culture supernatant. Reverse transcription-polymerase chain reaction(RT-PCR) was used to detect related mRNAs, and Western blot was used to detect the levels of iNOS and nuclear transcription factor NF-κBp65 at protein levels in cells. The results showed that ≤150 μg/mL high nucleotide yeast hydrolysates were safe for RAW264.7 cells. Compared with the LPS model with no intervention, 60-150 μg/mL high-nucleotide yeast hydrolysates could significantly enhance the phagocytic ability of RAW264.7 cells, inhibit the releases of NO, TNF-α, IL-1β, and IL-6 inflammatory factors and their related mRNAs in LPS-induced RAW264.7 cells. Besides, iNOS and NF-κB decreased at protein levels. The results confirmed the protective effects of high nucleotide yeast hydrolysates on LPS-induced inflammation in RAW264.7 cells, and the mechanism was related to the NF-κB pathway, which provides a theoretical basis for therapeutic intervention by foods for chronic diseases.
作者
潘聪
李占东
苑鹏
张大力
段盛林
夏凯
周文萱
赵可心
于伟厚
PAN Cong;LI Zhandong;YUAN Peng;ZHANG Dali;DUAN Shenglin;XIA Kai;ZHOU Wenxuan;ZHAO Kexin;YU Weihou(School of Food Science and Engineering,Jilin Agricultural University,Changchun 130118,China;China Food Fermentation Industry Research Institute,Beijing 100015,China;Dalian Shuangdi Technology,Dalian 116635,China;Beijing Key Laboratory of the Innovative Development of Functional Staple and the Nutritional Intervention forChronic Disease,Beijing 100015,China;School of Food Engineering,Jilin Engineering Normal University,Changchun 130052,China)
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2019年第7期8-14,共7页
Food and Fermentation Industries
基金
十三五-农机院薯类专项:薯类发酵主食新产品(面包)加工工艺技术研究(2016YFD0401303-02)
马铃薯主食加工装备研究及特膳食品开发(北京市科委重大专项)马铃薯主食化关键技术研究与产业化示范(D171100001917003)
关键词
酵母水解物
核苷酸
抗炎
NF-ΚB
yeast hydrolysate
nucleotide
anti-inflammatory
NF-κB
作者简介
第一作者:潘聪,硕士研究生;通讯作者:李占东,副教授,E-mail:lizd591@163.com.
