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壳寡糖激活巨噬细胞的机制 被引量:10

Mechanism of oligochitosan-induced macrophage activation
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摘要 目的:探讨巨噬细胞结合、内吞壳寡糖以及壳寡糖激活巨噬细胞的机制。方法:利用荧光物质2-氨基吖啶酮标记壳寡糖,在激光共聚焦显微镜下观察巨噬细胞内吞壳寡糖的过程,流式细胞仪分析细胞的荧光强度,通过竞争性抑制实验及钙离子、胰蛋白酶、秋水仙碱对内吞的影响来确定巨噬细胞内吞壳寡糖的机制。通过RT-PCR方法检测TNF-α来反映结合及内吞过程对壳寡糖刺激巨噬细胞的影响。结果:巨噬细胞可结合内吞壳寡糖。内吞易受温度影响,37℃较迅速(<2m in),4℃只能结合不能内吞。EDTA可抑制巨噬细胞内吞壳寡糖;经胰蛋白酶预处理的巨噬细胞摄取壳寡糖的能力大大降低,终止胰蛋白酶后巨噬细胞摄取壳寡糖的能力得到恢复。秋水仙碱预处理可明显抑制巨噬细胞摄取壳寡糖,并呈剂量依赖性。0.1 m o l/L甘露糖可抑制壳寡糖对巨噬细胞的刺激作用。结论:甘露糖受体介导的吞饮是巨噬细胞内吞壳寡糖的一条重要途径,甘露糖受体在壳寡糖激活巨噬细胞中起重要作用。 Objective: To study the mechanism of oligochitosan-induced macrophage activation. Methods: Oligochitosan was chemically modified with fluorophore 2-aminoacridone (2-AMAC). The cellulax events of 2-AMAC-oligochitosan-macrophage interaction were analyzed with confocal laser microscopy and the fluorescence intensity of cells was analyzed by BD LSR flow cytometer. The mechanism of oligochitosan uptake by macrophages was studied by competitive inhibition test and the effect of calcium,trypsin and colchicine on oligochitosan recognition and internalization were also determined. RT-PCR was performed to investigate the level of TNF-α secretion. Results: Macrophage could bind and uptake oligochitosan,which was dependent on the temperature: the uptake proceeded rapidly at 37 C and at 4 C macrophage could only bind oligochitosan. EDTA decreased oligochitosan uptake. Trypsin treatment significantly reduced the internalization,and uptake was recovered by trypsin termination. Colchicine significantly inhibited the internalization process and was dose dependent. 0. 1 mol/L mannose inhibited TNF-α expression induced by oligochitosan. Conclusions: Macrophage could uptake oligochitosan via mannose receptor mediated pinocytosis. Mannose receptor is crucial for the oligochitosan-induced macrophages activation.
出处 《浙江大学学报(医学版)》 CAS CSCD 2006年第3期265-272,共8页 Journal of Zhejiang University(Medical Sciences)
基金 浙江省自然科学基金(302031)
关键词 巨噬细胞 肿瘤坏死因子α 寡糖类/药理学 甘露糖 受体 细胞表面 胰蛋白酶/药理学 秋水仙碱/药理学 Macrophages Tumor necrosis factor-alpha Oligosaccharides/pharmacol Mannose Receptors ,cell surface Trypsin/pharmacol Colchicine/pharmacol
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参考文献19

  • 1PAE H O,SEO W G,KIM N Y,et al.Induction of granulocytic differentiation in acute promyelocytic leukemia cells (HL-60) by water-soluble chitosan oligomer[J].Leuk Res,2001,25(4):339-346.
  • 2SUZUKI K,MIKAMI T,OKAWA Y,et al.Antitumor effect of hexa-N-acetylchitohexaose and chitohexaose[J].Carbohyd Res,1987,151(15):403-408.
  • 3JEON Y J,PARK P J,KIM S K.Antimicrobial effect of chitooligosaccharides produced by biorector[J].Carbohydrate Polymers,2001,44(1):71-76.
  • 4FUJIWARA M,HAYASHI Y,OHARA N.Inhibitory effect of water-soluble chitosan on growth of Streptococcus mutans[J].New Microbiol,2004,27(1):83-86.
  • 5KWEON D K,SONG S B,PARK Y Y.Preparation of water-soluble chitosan/heparin complex and its application as wound healing accelerator[J].Biomaterials,2003,24(9):1 595-1 601.
  • 6竺国芳,赵鲁杭.几丁寡糖和壳寡糖的研究进展[J].中国海洋药物,2000,19(1):43-46. 被引量:78
  • 7SHIBATA Y,FOSTER L A,METZGER W J,et al.Alveolar macrophage priming by intravenous administration of chitin particles,polymers of N-acetyl-D-Glucosamine,in mice[J].Infection and Immunity,1997,65(5):1 734-1 741.
  • 8MUZZARELLI R A A.Human enzymatic activities related to the therapeutic administration of chitin derivatives[J].Cell Mol Life Sci,1997,53(12):131-140.
  • 9NISHIMURA K,ISHIHARA C,UKEI S,et al.Stimulation of cytokine production in mice using deacetylated chitin[J].Vaccine,1986,4(3):151-156.
  • 10纪莹,赵轶,周翔,翁登坡,赵鲁杭.壳寡糖的制备及组分分析[J].中国现代应用药学,2003,20(3):195-196. 被引量:16

二级参考文献10

  • 1金城,张树政.糖生物学与糖工程的兴起与前景[J].生物工程进展,1995,15(3):12-17. 被引量:32
  • 2章莹.几丁质应用于肿瘤治疗的研究现状[J].国外医学(肿瘤学分册),1996,23(6):346-348. 被引量:25
  • 3Shibata Y, Foster LA, Metzger WJ, et al. Alveolar macrophage priming by intravenous administration of chitin particles, polymers of N-acetyl-D-glucosamine, in mice [J]. Infect Immun 1997,65(5): 1734.
  • 4Shibata Y, Foster LA, Bradfield JF, et al. Oral administration of chitin down-regulates serum IgE levels and lung eosinophilia in the allergic mouse[J]. J Immunol 2000; 164(3) : 1314.
  • 5Shibata Y, Honda I,Justice JP, et al. The adjuvant N-acetyl-D-glucosamine polymer up-regulates Th1 immunity but downregulates Th2 immunity against a mycobacterial protein(MPB-59)in interleukin-10-knockout and wild-type mice[J].Infect Immun 2001 ; 69(10) : 6123.
  • 6Suzuki K, Mikami T, Okawa Y, et al. Antitumor effect of hexa-N-acetylchitohe-xaose and chitohexaose[J]. Carbohydr Res 1986; 151:403.
  • 7Tsukada K, Matsumoto T, Aizawa K,et al. Antimetastatic and growth-inhibitory effects of N-acetylchitohexaose in mice beating Lewis lung carcinoma[J].Jpn J Cancer Res 1990; 81(3) : 259.
  • 8王中和,陆顺娟,胡海生,张澄波.低分子壳多糖对癌症放疗患者免疫功能的影响[J].首都医科大学学报,1997,18(1):80-82. 被引量:29
  • 9杜昱光,白雪芳,虞星炬,韩秀文.寡聚糖类物质生理活性的研究[J].中国生化药物杂志,1997,18(5):268-270. 被引量:71
  • 10张虎,杜昱光,虞星炬.几丁寡糖与壳寡糖的制备和功能[J].中国生化药物杂志,1999,20(2):99-101. 被引量:50

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