中国动脉硬化杂志2017，Vol.25Issue(11)：1093-1098,1106,7.
原核表达Lunasin的抗炎生物学活性鉴定及其对J774A.1细胞组织因子表达的影响
Identification of anti-inflammatory biological activity of prokaryotic expressed lunasin and its effect on the expression of tissue factor in J774A.1 cells
摘要
Abstract
Aim To test the anti-inflammatory biological activity of prokaryotic expressed lunasin,and to lay the foundation for future lunasin development in biomedical field.Methods The lunasin DNA fragment amplified by PCR was cloned into the expression vector pET28 by means of genetic engineering technology.After the identification by restriction enzyme digestion and sequencing,lunasin expression was induced.The purification of lunasin was performed by nickel ion affinity chromatography.Griess method and ELISA were used to test the effects of lunasin on the contents of nitric oxide,tumor necrosis factor-α and interleukin-6 in the culture medium of mouse J774A.1 cells.The effects of lunasin on the phosphorylation of nuclear factor-κB (NF-κB) p65 and the expression of tissue factor (TF) were detected by Western blot.Results Prokaryotic expression vector of lunasin was successfully constructed and the recombinant lunasin peptide with about 90％ purification was obtained.Lunasin could inhibit activation of NF-κB induced by lipopolysaccharide (LPS) in mouse J774A.1 cells and then subsequently suppressed the synthesis of nitric oxide and the expression of downstream genes like tumor necrosis factor-α and interleukin-6.Lunasin also significantly inhibited the expression of TF induced by LPS in J774A.1 cells.Conclusion Prokaryotic expressed lunasin shows significant anti-inflammatory biological activity,and significantly inhibits the TF expression induced by LPS in J774A.1 cells,and its mechanism may be achieved by inhibiting the activation of NF-κB.关键词
Lunasin/核因子κB/炎症因子/组织因子/脂多糖Key words
Lunasin/Nuclear factor-κB/Inflammatory cytokine/Tissue factor/Lipopolysaccharide分类
医药卫生引用本文复制引用
陈玉华,付常振,李明英,李敬达,迟彦..原核表达Lunasin的抗炎生物学活性鉴定及其对J774A.1细胞组织因子表达的影响[J].中国动脉硬化杂志,2017,25(11):1093-1098,1106,7.基金项目
国家自然科学基金资助项目(81202536) （81202536）
大连市科学计划项目(2015E21SF012) （2015E21SF012）
