Application of DNA tetrahedron in the aspect of improving anti-aging process of cells

A tetrahedral, anti-aging technology, used in medical preparations containing active ingredients, anti-inflammatory agents, anti-toxic agents, etc., to achieve good bioavailability, delay aging, and inhibit expression.

Active Publication Date: 2018-04-27
常州智胜生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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But there is no report on the application of DNA tetrahedron in anti-inflammation and anti-oxidation of cells

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  • Application of DNA tetrahedron in the aspect of improving anti-aging process of cells
  • Application of DNA tetrahedron in the aspect of improving anti-aging process of cells
  • Application of DNA tetrahedron in the aspect of improving anti-aging process of cells

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Experimental program
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Embodiment 1

[0031] Synthesis and characterization of embodiment 1 DNA tetrahedron

[0032] 1. Synthesis of DNA tetrahedron

[0033] Add the four specially designed single strands of the DNA tetrahedron into a 200μl EP tube containing 100ul TM buffer (10mMTris-HCl, 50mM MgCl2, pH 8.0) in an equimolar ratio, and then maintain the mixed system at 95°C for 10min, then rapidly cool down Keep it at 4°C for 20 minutes to synthesize a DNA tetrahedron, which can be stored at 4°C for a long time.

[0034] The specific sequences of the four DNA single strands are as follows:

[0035] S1:

[0036] 5'-ATTTATTCACCCGCCATAGTAGACGTATCACCAGGCAGTTGAGACGAACATTCCTAAGTCTGAA-3' (SEQ ID NO: 1);

[0037] S2:

[0038] 5'-ACATGCGAGGGTCCAATACCGACGATTACAGCTTGCTACACGATTCAGACTTAGGAATGTTCG-3' (SEQ ID NO: 2);

[0039] S3:

[0040] 5'-ACTACTATGGCGGGTGATAAAACGTGTAGCAAGCTGTAATCGACGGGAAGAGCATGCCCATCC-3' (SEQ ID NO: 3);

[0041] S4:

[0042] 5'-ACGGTATTGGACCCTCGCATGACTCAACTGCCTGGTGATACGAGGATGGGCATGCTCTTCCCG-3' (SEQ ID...

Embodiment 2G

[0048] Example 2 Griess method to detect the influence of DNA tetrahedron on secretion of inflammatory factor NO by inflammatory model cells

[0049]The biological effects of NO depend on its concentration and subtle changes inside and outside cells. Under normal physiological conditions, a small amount of NO is beneficial to the body. However, high concentrations of NO can cause tissue damage, malignancy, rheumatoid arthritis, and septic shock. To investigate whether TDNs play a role in inflammatory response, we treated RAW264.7 cells with lipopolysaccharide (LPS) to establish an inflammatory response model. Production of the inflammatory mediator NO was examined by Griess assay.

[0050] Experimental method: The experiment is divided into four groups, namely: negative control group, positive control group, experimental group 1, and experimental group 2.

[0051] First, RAW264.7 cells were inoculated in 96-well cell culture plates, cultured in medium containing 10% FBS for...

Embodiment 3

[0053] Example 3 Study on the anti-inflammatory mechanism of DNA tetrahedron on RAW264.7 cells

[0054] Endogenous NO is mainly derived from L-arginine, and three isoenzymes can convert L-arginine to L-citrulline and NO, namely, neuronal NO synthase, endothelial NO synthase, and inducible NO synthase (iNOS)31,32. iNOS is widely distributed in the human body, especially in macrophages, hepatocytes, astrocytes and smooth muscle cells. Therefore, we further explored the effects of TDNs on iNOS levels by detecting the expression of iNOS gene and protein by confocal microscopy, Western Blotting, and fluorescent quantitative PCR.

[0055] experiment method:

[0056] (1) The experiment is divided into four groups, namely: negative control group, positive control group, experimental group 1, and experimental group 2.

[0057] First, RAW264.7 cells were seeded in 96-well cell culture plates and cultured in 10% FBS at 37°C, 5% CO 2 Cultivate in the incubator for 8h, then change to t...

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Abstract

The invention provides an application of a DNA tetrahedron in the aspect of improving anti-aging process of cells. The DNA tetrahedron is synthesized from four DNA single chains in a self-assembling manner, wherein the sequences of the four single chains are represented as the SEQ ID No.1-4. The anti-aging process of cells includes anti-inflammation and anti-oxidizing processes, wherein the anti-inflammation effect, improved by the DNA tetrahedron, of the cells is achieved by reducing release of inflammatory cytokines and inhibiting expression quantities of iNOS gene and iNOS protein; the anti-oxidizing of the cells is achieved by inhibiting the phosphorylation of proteins ERK1/2, P38 and JNK at the downstream of MAPK/ERK signal channel, and improving the expression quantity of inducing heme oxygenase-1. The DNA tetrahedron has excellent bioavailability and biocompatibility, has anti-inflammation and anti-oxidizing functions and further delays senility of cells.

Description

technical field [0001] The invention belongs to the technical field of cell anti-inflammation and anti-oxidation, and specifically relates to the application of a DNA tetrahedron in promoting cell anti-aging. Background technique [0002] With the rapid development of DNA nanotechnology, various DNA nanostructures have been fabricated. DNA tetrahedral (tetrahedral DNA nanostructures, TDNs), as one of the most promising DNA nanostructures, has recently been considered as a biomaterial with a wide range of potential applications. TDNs are self-assembled from four highly specific and programmable single-stranded DNA (ssDNA). The design of single-stranded DNA has precise complementary base pairing, sequence design and specific hybridization of complementary functional sequences. Unlike ssDNA, TDNs can enter mammalian cells without lipofection and remain intact in the cytoplasm for at least 48 hours, an important property for drug delivery and control of their activity in cells...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K31/7088A61P39/06A61P29/00
CPCA61K31/7088A61K48/005
Inventor 林云锋张琦蔡潇潇林世宇田陶然马全诠张陶
Owner 常州智胜生物科技有限公司
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