Construction of glucosamine/DNA composite nanomaterials and its effect on function of Raw264.7 cells

Yuhang XU; Yue CHEN; Qing XIANG; Shuoxin ZHANG; Daohui GONG; Di WU; Guansong WANG; Hang QIAN

doi:10.16016/j.2097-0927.202310033

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Construction of glucosamine/DNA composite nanomaterials and its effect on function of Raw264.7 cells

Basic Medicine | 更新时间：2026-01-09

- Construction of glucosamine/DNA composite nanomaterials and its effect on function of Raw264.7 cells
- Journal of Army Medical University Vol. 46, Issue 13, Pages: 1494-1501(2024)
- 作者机构：
  
  400037 重庆，陆军军医大学(第三军医大学)第二附属医院呼吸与危重症医学科
- 作者简介：
- 基金信息：
  
  the General Program of Natural Science Foundation in Chognqing(cstc2021jcyj-msxmX1179)
- DOI：10.16016/j.2097-0927.202310033
  CLC： R914.2;R967;R977.6
- Received：08 October 2023，
  
  Revised：2023-12-01，
  
  Published：15 July 2024
- 稿件说明：
移动端阅览
Yuhang XU, Yue CHEN, Qing XIANG, et al. Construction of glucosamine/DNA composite nanomaterials and its effect on function of Raw264.7 cells[J]. Journal of Army Medical University, 2024, 46(13): 1494-1501.
DOI：

Yuhang XU, Yue CHEN, Qing XIANG, et al. Construction of glucosamine/DNA composite nanomaterials and its effect on function of Raw264.7 cells[J]. Journal of Army Medical University, 2024, 46(13): 1494-1501. DOI： 10.16016/j.2097-0927.202310033.

摘要

目的

构建一种无镁离子组装的新型氨基葡萄糖/DNA复合纳米材料，验证氨基葡萄糖能否介导组装DNA纳米管(nanotube，NT)，并评价其对体外Raw264.7细胞功能的影响。

方法

采用梯度退火法，使用Y1、Y2和Y3这3条DNA单链，以氨基葡萄糖(glucosamine，GlcN)为介质组装DNA NT，构建氨基葡萄糖/DNA复合纳米材料(NT

GlcN

)。使用原子力显微镜(atomic force microscopy，AFM)观察其纳米结构，激光动态光散射(dynamic light scattering，DLS)测量该纳米材料的尺寸。选用Raw264.7细胞系，采用CCK-8法测试氨基葡萄糖或NT

GlcN

的细胞毒性，流式细胞术和激光共聚焦考察细胞对该纳米结构的摄取效率，RT-qPCR测定NT

GlcN

和NT

(镁离子介导组装的DNA纳米管)对脂多糖(lipopolysaccharides，LPS)诱导的巨噬细胞炎症因子(IL-1β、IL-6)表达水平的影响。

结果

氨基葡萄糖可以成功介导NT

GlcN

的合成，稳定性较好。AFM表征结果表明，NT

GlcN

为管状颗粒，均匀地分散在云母表面上；DLS测量表明NT

GlcN

的直径为(15.26±3.86) nm。细胞实验显示：较之NT

，巨噬细胞对NT

GlcN

有更高的细胞摄取效率，NT

GlcN

处理后细胞存活率更高(

＜0.05)；NT

GlcN

处理后，LPS诱导的巨噬细胞炎症因子表达降低(

＜0.05)。

结论

本研究构建的氨基葡萄糖/DNA复合纳米材料具有良好的稳定性、生物相容性和细胞摄取效率；能降低氨基葡萄糖的细胞毒性，并能通过降低Raw 264.7细胞炎症因子的表达来抑制细胞炎症反应。

Abstract

Objective

To construct a new type of glucosamine/DNA composite nanostructure (NT

GlcN

) assembled without magnesium

verify whether or not glucosamine can mediate the assembly of DNA nanotubes (NT) and assess its effect on the function of Raw264.7 cells.

Methods

Utilizing the gradient annealing method with 3 DNA single strands Y1

and Y3

glucosamine (GlcN) was employed to mediate the assembly of DNA NT

resulting in the formation of glucosamine/DNA composite nanostructures. Atomic force microscopy (AFM) was used to observe the surface structure of the nanomaterial and dynamic light scattering (DLS) was used to measure its size. RAW264.7 cells were used in cell experiments. The cytotoxicity of GlcN and NT

GlcN

was assessed using CCK-8 assay. Flow cytometry and laser confocal microscopy were employed to investigate the cellular uptake efficiency of the nanostructures. The effects of NT

GlcN

and NT

(Mg

-assembled of DNA NT) on the expression levels of inflammatory cytokines (IL-1β

IL-6) in macrophages induced by lipopolysaccharides (LPS) were evaluated using RT-qPCR.

Results

GlcN successfully mediated the synthesis of NT

GlcN

which exhibited good stability. AFM characterization results revealed that NT

GlcN

formed tubular particles that were uniformly distributed on the surface of mica. DLS measurements indicated that the diameter of NT

GlcN

was approximately 15.26±3.86 nm. Cell experiments demonstrated that

compared to NT

macrophages exhibited a higher cellular uptake efficiency for NT

GlcN

with a higher cell survival rate following treatment with NT

GlcN

(

0.05). After NT

GlcN

treatment

the expression of inflammatory cytokines in LPS-induced macrophages was reduced (

0.05).

Conclusion

The glucosamine/DNA composite nanostructures have been successfully developed

possessing excellent stability

biocompatibility and cell uptake efficiency. NT

GlcN

is capable of reducing the cytotoxicity of GlcN and can suppress cellular inflammatory responses by decreasing the expression of inflammatory cytokines in RAW264.7 cells.

关键词

Keywords

references

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