浏览全部资源
扫码关注微信
氨基葡萄糖/DNA复合纳米材料的构建及其对Raw264.7细胞功能的影响
Construction of glucosamine/DNA composite nanomaterials and its effect on function of Raw264.7 cells
- 陆军军医大学学报 2024年46卷第13期 页码：1494-1501
- 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：
[钱航，E-mail: hqian@tmmu.edu.cn QIAN Hang, E-mail: hqian@tmmu.edu.cn
- 基金信息：重庆市自然科学基金面上项目(cstc2021jcyj-msxmX1179)
DOI：10.16016/j.2097-0927.202310033
中图分类号： R914.2;R967;R977.6收稿：2023-10-08，
修回：2023-12-01，
纸质出版：2024-07-15
- 稿件说明：
移动端阅览
徐宇航, 陈跃, 向青, 等. 氨基葡萄糖/DNA复合纳米材料的构建及其对Raw264.7细胞功能的影响[J]. 陆军军医大学学报, 2024,46(13):1494-1501.
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.
徐宇航, 陈跃, 向青, 等. 氨基葡萄糖/DNA复合纳米材料的构建及其对Raw264.7细胞功能的影响[J]. 陆军军医大学学报, 2024,46(13):1494-1501. DOI： 10.16016/j.2097-0927.202310033.
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
2
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
2
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
Mg
(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
2
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
Mg
，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(
P
＜0.05)；NT
GlcN
处理后，LPS诱导的巨噬细胞炎症因子表达降低(
P
＜0.05)。
结论
2
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
2
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
2
Utilizing the gradient annealing method with 3 DNA single strands Y1
Y2
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
(Mg
2+
-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
2
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
Mg
macrophages exhibited a higher cellular uptake efficiency for NT
GlcN
with a higher cell survival rate following treatment with NT
GlcN
(
P
<
0.05). After NT
GlcN
treatment
the expression of inflammatory cytokines in LPS-induced macrophages was reduced (
P
<
0.05).
Conclusion
2
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
QIAO Q, LIU X, YANG T, et al . Nanomedicine for acute respiratory distress syndrome: the latest application, targeting strategy and rational design[J ] . Acta Pharm Sin B, 2021, 11(10): 3060-3091. DOI: 10.1016/j.apsb.2021.04.023.
MELA I, VALLEJO-RAMIREZ P P, MAKARCHUK S, et al . DNA nanostructures for targeted antimicrobial delivery[J ] . Angew Chem Int Ed Engl, 2020, 59(31): 12698-12702. DOI: 10.1002/anie.202002740.
ZHANG Y X, MA W J, ZHU Y, et al . Inhibiting methicillin-resistant Staphylococcus aureus by tetrahedral DNA nanostructure-enabled antisense peptide nucleic acid delivery[J ] . Nano Lett, 2018, 18(9): 5652-5659. DOI: 10.1021/acs.nanolett.8b02166..
YANG L, YAO C, LI F, et al . Synthesis of branched DNA scaffolded super-nanoclusters with enhanced antibacterial performance[J ] . Small, 2018, 14(16): e1800185. DOI: 10.1002/smll.201800185.
HU Q Q, LI H, WANG L H, et al . DNA nanotechnology-enabled drug delivery systems[J ] . Chem Rev, 2019, 119(10): 6459-6506. DOI: 10.1021/acs.chemrev.7b00663.
KELLER A, LINKO V. Challenges and perspectives of DNA nanostructures in biomedicine[J]. Angew Chem Int Ed Engl, 2020, 59(37): 15818-15833. DOI: 10.1002/anie.201916390.
LU X H, LIU J B, WU X H, et al . Multifunctional DNA origami nanoplatforms for drug delivery[J ] . Chem Asian J, 2019, 14(13): 2193-2202. DOI: 10.1002/asia.201900574.
KELLY G S. The role of glucosamine sulfate and chondroitin sulfates in the treatment of degenerative joint disease[J]. Altern Med Rev, 1998, 3(1): 27-39.
WU Y L, LIN A H, CHEN C H, et al . Glucosamine attenuates cigarette smoke-induced l ung inflammation by inhibiting ROS-sensitive inflammatory signaling[J ] . Free Radic Biol Med, 2014, 69: 208-218. DOI: 10.1016/j.freeradbiomed.2014.01.026.
XING R E, LIU S, GUO Z Y, et al . The antioxidant activity of glucosamine hydrochloride in vitro [J ] . Bioorg Med Chem, 2006, 14(6): 1706-1709. DOI: 10.1016/j.bmc.2005.10.018..
MENDIS E, KIM M M, RAJAPAKSE N, et al . Sulfated glucosamine inhibits oxidation of biomolecules in cells via a mechanism involving intracellular free radical scavenging[J ] . Eur J Pharmacol, 2008, 579(1/2/3): 74-85. DOI: 10.1016/j.ejphar.2007.10.027..
VALVASON C, MUSACCHIO E, POZZUOLI A, et al . Influence of glucosamine sulphate on oxidative stress in human osteoarthritic chondrocytes: effects on HO-1, p22(Phox) and iNOS expression[J ] . Rheumatology, 2008, 47(1): 31-35. DOI: 10.1093/rheumatology/kem289.
RAJAPAKSE N, MENDIS E, KIM M M, et al. Sulfated glucosamine inhibits MMP-2 and MMP-9 expressions in human fibrosarcoma cells[J]. Bioorg Med Chem, 2007, 15(14): 4891-4896. DOI: 10.1016/j.bmc.2007.04.048
YANG Y, LIU W S, HAN B Q, et al . The antioxidative and immunostimulating properties of D-glucosamine[J ] . Int Immunopharmacol, 2007, 7(1): 29-35. DOI: 10.1016/j.intimp.2006.06.003.
ARDEN N K, PERRY T A, BANNURU R R, et al . Non-surgical management of knee osteoarthritis: comparison of ESCEO and OARSI 2019 guidelines[J ] . Nat Rev Rheumatol, 2021, 17(1): 59-66. DOI: 10.1038/s41584-020-00523-9.
CHUANG K H, PENG Y C, CHIEN H Y, et al . Attenuation of LPS-induced lung inflammation by glucosamine in rats[J ] . Am J Respir Cell Mol Biol, 2013, 49(6): 1110-1119. DOI: 10.1165/rcmb.2013-0022OC.
HWANG J S, KIM K H, PARK J, et al . Glucosamine improves survival in a mouse model of sepsis and attenuates sepsis-induced lung injury and inflammation[J ] . J Biol Chem, 2019, 294(2): 608-622. DOI: 10.1074/jbc.RA118.004638.
WU Y L, KOU Y R, OU H L, et al . Glucosamine regulation of LPS-mediated inflammation in human bronchial epithelial cells[J ] . Eur J Pharmacol, 2010, 635(1/2/3): 219-226. DOI: 10.1016/j.ejphar.2010.02.044.
YOMOGIDA S, KOJIMA Y, TSUTSUMI-ISHII Y, et al . Glucosamine, a naturally occurring amino monosaccharide, suppresses dextran sulfate sodium-induced colitis in rats[J ] . Int J Mol Med, 2008, 22(3): 317-323.
HWANG S Y, SHIN J H, HWANG J S, et al . Glucosamine exerts a neuroprotective effect via suppression of inflammation in rat brain ischemia/reperfusion injury[J ] . Glia, 2010, 58(15): 1881-1892. DOI: 10.1002/glia.21058.
WANG D, LIU Q, WU D, et al . Isothermal self-assembly of spermidine-DNA nanostructure complex as a functional platform for cancer therapy[J ] . ACS Appl Mater erfaces, 2018, 10(18): 15504-15516. DOI: 10.1021/acsami.8b03464.
HUANG C W, YOU Q Y, XUJ, et al . An mTOR siRNA-loaded spermidine/DNA tetrahedron nanoplatform with a synergistic anti-inflammatory effect on acute lung injury[J ] . Adv Healthc Mater, 2022, 11(11): e2200008. DOI: 10.1002/adhm.202200008.
WANG D, CHEN C F, LIU Q, et al . Functionalizing DNA nanostructures with natural cationic amino acids[J ] . Bioact Mater, 2021, 6(9): 2946-2955. DOI: 10.1016/j.bioactmat.2021.02.012.
CHEN C F, LIU Q, YUAN Y, et al . Protein-mediated DNA self-assembly by controlling the surface charge in a molecular crowding environment[J ] . Biomater Sci, 2022, 10(8): 2006-2013. DOI: 10.1039/D1BM02017J.
WANG C, VÁZQUEZ-GONZÁLEZ M, FADEEV M, et al . Thermoplasmonic-triggered release of loads from DNA-modified hydrogel microcapsules functionalized with Au nanoparticles or Au nanorods[J ] . Small, 2020, 16(22): e2000880. DOI: 10.1002/smll.202000880.
MIKI K, ORIDE K, KIMURA A, et al . Influence of side chain length on fluorescence intensity of ROMP-based polymeric nanoparticles and their tumor specificity in in-vivo tumor imaging[J ] . Small, 2011, 7(24): 3536-3547. DOI: 10.1002/smll.201101637..
0
浏览量
139
下载量
0
CSCD
- 网络PDF下载
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
