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通过 miRNA-382 研究 ssDNA 单壁碳纳米管的不同结合和细胞内递药效率及其对肾系膜细胞中 LC3 相关自噬的影响。

Studying Different Binding and Intracellular Delivery Efficiency of ssDNA Single-Walled Carbon Nanotubes and Their Effects on LC3-Related Autophagy in Renal Mesangial Cells via miRNA-382.

机构信息

Division of Nephrology, Nanfang Hospital, Southern Medical University , Key Lab for Organ Failure Research, Ministry of Education, Guangzhou, Guangdong 510515, China.

Department of Anatomy, Guangdong Provincial Key Laboratory of Construction and Detection in Tissue Engineering, Southern Medical University , Guangdong, Guangzhou 510515, China.

出版信息

ACS Appl Mater Interfaces. 2015 Nov 25;7(46):25733-40. doi: 10.1021/acsami.5b07185. Epub 2015 Nov 16.

Abstract

Single-walled carbon nanotubes (SWCNTs) have been used to deliver single-stranded (ssDNA). ssDNA in oligonucleotide can act as an inhibitor of microRNA to regulate cellular functions. However, these ssDNA are difficult to bind carbon nanotubes with low transferring efficiency to cells. To this end, we designed ssDNA with regulatory and functional units to form ssDNA-SWCNT hybrids to study their binding effects and transferring efficiency. The functional unit on ssDNA mimics the inhibitor (MI) of miRNA-382, which plays a crucial role in the progress of many diseases such as renal interstitial fibrosis. After verification of overexpression of miRNA-382 in a coculture system, we designed oligonucleotide sequences (GCG)5-MI, (TAT)5-MI, and N23-MI as regulatory units added to the 5'-terminal end of the functional DNA fragment, respectively. These regulatory units lead to different secondary structures and thus exhibit different affinity ability to SWCNTs, and finally decide their deliver efficacy to cells. Autophagy, apoptosis and necrosis were observed in renal mesangial cells.

摘要

单壁碳纳米管 (SWCNTs) 已被用于输送单链 (ssDNA)。寡核苷酸中的 ssDNA 可以作为 microRNA 的抑制剂,调节细胞功能。然而,这些 ssDNA 与碳纳米管的结合能力较弱,向细胞的转移效率也较低。为此,我们设计了带有调节和功能单元的 ssDNA,以形成 ssDNA-SWCNT 杂交体,研究它们的结合效果和转移效率。ssDNA 上的功能单元模拟 miRNA-382 的抑制剂 (MI),miRNA-382 在许多疾病的进展中起着至关重要的作用,如肾间质纤维化。在共培养系统中验证了 miRNA-382 的过表达后,我们分别设计了寡核苷酸序列 (GCG)5-MI、(TAT)5-MI 和 N23-MI 作为调节单元,添加到功能 DNA 片段的 5'-末端。这些调节单元导致不同的二级结构,从而表现出与 SWCNTs 不同的亲和力,最终决定了它们向细胞的递送效率。观察到肾小球系膜细胞中的自噬、凋亡和坏死。

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