黑豆纳豆激酶粗提取物的免疫调节作用  

The Immunomodulatory Effects of Black Bean Nattokinase Crude Extract

作　　者：田玥玮 张永坡[1] 高春艳[1] 赵晋忠[1] 杜维俊[2] Tian Yuewei;Zhang Yongpo;Gao Chunyan;Zhao Jinzhong;Du Weijun(Department of Basic Sciences,Shanxi Agricultural University,Jinzhong 030801,Shanxi;College of Agronomy,Shanxi Agricultural University,Jinzhong 030801,Shanxi)

机构地区：[1]山西农业大学基础部,山西晋中030801 [2]山西农业大学农学院,山西晋中030801

出　　处：《中国食品学报》2024年第1期31-40,共10页Journal of Chinese Institute Of Food Science and Technology

基　　金：山西省重点研发计划项目(201903D221036,201903D2110061-1);国家重点研发计划项目(2021YFD1600601-2);中央引导地方科技发展资金项目(YDZJSX2022A035)。

摘　　要：以壶关大黑豆为原料发酵纳豆,分离得到黑豆纳豆激酶粗提取物(BNCE),对其结构进行初探,测定其酶活力和溶栓能力,并研究BNCE的免疫调节作用。通过红外光谱分析和扫描电镜对BNCE结构进行鉴定;分别使用纤维蛋白平板法和动物血块法测定BNCE的酶活及其对小鼠血块的体外溶栓作用;用MTT比色法探究BNCE对RAW264.7细胞增殖的影响;中性红染色法测定BNCE对RAW264.7细胞吞噬率的影响;ELISA法检测BNCE对RAW264.7细胞分泌IL-6、TNF-α和IL-1β的影响。结果表明,BNCE具有蛋白质的化学结构,表观呈现大小不同且无规则的碎片形状,具有一定的纤溶活性和体外溶栓作用。BNCE可以促进正常状态以及LPS激活状态下RAW264.7细胞的增殖,增强其吞噬率,并且有浓度依赖性。另外,BNCE还可以促进正常状态下的RAW264.7细胞TNF-α和IL-1β的分泌,抑制IL-6的分泌;也能抑制LPS激活状态下RAW264.7细胞IL-6、TNF-α和IL-1β的分泌。因此,BNCE具有一定的免疫调节作用,在食品工业上可以作为保健食品的一种优质原料。In this paper,Black bean nattokinase crude extract(BNCE)was isolated by fermenting natto from Huguan black beans.Its structural was preliminary investigated,as well as the determination of enzyme activity,thrombolytic function and immunomodulatory effect were carried out.The structure of BNCE is identified by infrared spectroscopy and scanning electron microscopy.Enzymatic activity and thrombolytic function of BNCE were determined by fibrin plate method and animal blood clot method,respectively.The effect of BNCE on the proliferation of RAW264.7 cells was investigated by MTT colorimetric method.Neutral red staining was employed to investigate the effect of BNCE on the phagocytosis rate of RAW264.7 cells.In the effect of BNCE,the secretion of IL-6,TNF-αand IL-1βby RAW264.7 cells was investigated by ELISA.The results showed that BNCE was a protein with different sizes and irregular shapes of fragments,exhibiting certain fibrinolytic activity and thrombolytic function.BNCE can promote the proliferation and enhanced the phagocytosis rate of RAW264.7 cells in normal state as well as in the LPS-activated state in a concentration-dependent manner.In addition,BNCE can also promote the secretion of TNF-αand IL-1βand inhibite the secretion of IL-6 in normal state RAW264.7 cells.Meanwhile,BNCE can also inhibite the secretion of IL-6,TNF-αand IL-1βin LPS-activated state RAW264.7 cells.Therefore,BNCE has certain immunomodulatory effects,and can be used as a good raw material for food industry.

关 键 词：黑豆 纳豆激酶 溶栓作用 RAW264.7细胞 免疫调节 

分 类 号：TS214[轻工技术与工程—粮食、油脂及植物蛋白工程]