食品科学 ›› 2019, Vol. 40 ›› Issue (23): 170-175.doi: 10.7506/spkx1002-6630-20180930-337

• 营养卫生 • 上一篇    下一篇

重组黑芝免疫调节蛋白对小鼠巨噬细胞RAW264.7向M1型分化的影响

刘玲,李奇璋,周选围,王玉亮   

  1. (上海交通大学农业与生物学院,上海 200240)
  • 出版日期:2019-12-15 发布日期:2019-12-24
  • 基金资助:
    上海市自然科学基金项目(18ZR1420600);国家卫生健康委员会重大新药创制科技重大专项(2017ZX09101002-003-002)

Effect of Recombinant Ganoderma atrum Immunomodulatory Protein on Macrophage RAW264.7 Differentiation into M1 Type Cells

LIU Ling, LI Qizhang, ZHOU Xuanwei, WANG Yuliang   

  1. (School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China)
  • Online:2019-12-15 Published:2019-12-24

摘要: 真菌免疫调节蛋白具有免疫调节和抗肿瘤功效。巨噬细胞在肿瘤免疫治疗中发挥重要的作用,M1型巨噬细胞具有很强的肿瘤杀伤能力和抗原递呈能力。本研究通过真菌免疫调节蛋白在体外对小鼠巨噬细胞RAW264.7经典活化M1型的影响探究其抗肿瘤的作用机制。利用重组黑芝免疫调节蛋白(recombinant Ganoderma atrum immunomodulatory protein,rFIP-gat)干预RAW264.7细胞,用中性红吞噬实验测定细胞吞噬能力,一氧化氮(nitric oxide,NO)检测试剂盒测定NO浓度,采用实时荧光定量聚合酶链式反应测定肿瘤坏死因子-α(tumor necrosis factor α,TNF-α)、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)的mRNA相对表达量。结果表明,rFIP-gat以剂量依赖的方式增强RAW264.7细胞的吞噬能力,促进其产生NO,并提高iNOS和TNF-α基因的转录。因此,rFIP-gat可能具有诱导RAW264.7细胞向M1型巨噬细胞分化的作用。

关键词: 重组黑芝免疫调节蛋白, 巨噬细胞RAW264.7, 肿瘤坏死因子-α, 诱导型一氧化氮合酶

Abstract: Fungal immunomodulatory proteins (FIPs) have immunomodulatory and anti-tumor effects. Macrophages play an important role in tumor immunotherapy, and M1 type macrophages have strong tumor killing ability and antigen presentation ability. This study aimed to explore the effect and underlying mechanism of FIPs on the in vitro activation of macrophage RAW264.7 cells into M1 type. After being treated with a recombinant Ganoderma atrum immunomodulatory protein (rFIP-gat), the phagocytosis ability of RAW264.7 cells was determined by neutral red uptake assay, and nitric oxide (NO) production by a nitric oxide detection kit. Quantitative real-time polymerase chain reaction (qPCR) was used to determine the relative expression levels of tumor necrosis factor-α (TNF-α) and inducible nitric oxide synthase (iNOS) genes. The results indicated that rFIP-gat enhanced phagocytosis, promoted the production of nitric oxide, and increased the transcription of iNOS and TNF-α in macrophage RAW264.7 cells in a dose-dependent manner. In conclusion, rFIP-gat may induce the differentiation of macrophage RAW264.7 cells into M1 type.

Key words: recombinant Ganoderma atrum immunomodulatory protein, macrophage RAW264.7, tumor necrosis factor-α, inducible nitric oxide synthase

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