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Screening method for pharmaceutical composition and inhibitor for hippocampal function decline

A screening method, hippocampal technology, applied in drug combinations, biochemical equipment and methods, pharmaceutical formulations, etc.

Active Publication Date: 2021-06-11
DAEGU GYEONGBUK INST OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] In the prior art, it has been reported that microRNAs in the whole brain of mice are potentially involved in the aging process through the insulin signaling pathway (Sachi Inukai et al., PLoS ONE, 7(7):e40028, 2012), and it is known that The regulation of neural activity is based on the interaction of their downstream (down stream) targets in the microRNAs and specific biological pathways of the hippocampus (Stephen M.Eacker et al., PLoS ONE, 6(10): e25068, 2011), but in fact, there is a lack of comprehensive studies on how the dynamic expression of microRNAs age-dependently affects normal hippocampal function

Method used

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  • Screening method for pharmaceutical composition and inhibitor for hippocampal function decline
  • Screening method for pharmaceutical composition and inhibitor for hippocampal function decline
  • Screening method for pharmaceutical composition and inhibitor for hippocampal function decline

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0100] Example 1. Profile analysis of small RNAs (small RNAs) related to the aging process of the hippocampus

[0101] In the present invention, in order to understand the microRNAs that mediate the regulation of the aging process of the hippocampus, small RNA (small RNA; smRNA or sRNA, hereinafter referred to as sRNA) was performed on male mice of 2 months, 6 months, and 18 months. analyze.

[0102] 1-1: Animal preparation and tissue collection

[0103] Three common wild animals of different ages at 2 months, 6 months and 18 months were obtained from the Korea Research Institute of Bioscience & Biotechnology-Institutional animal care and use committee (KRIBB-IACUC). type male C57BL / 6J mice.

[0104] Hippocampus tissue was immediately dissected and cryopreserved using liquid nitrogen by sacrificing normal mice without tumors or traces of disease. Their health status was confirmed by a series of serum tests, and eight hippocampal tissues were obtained from four mice in eac...

Embodiment 2

[0132] Example 2. Analysis of the pathway of the target gene

[0133] All catalogs of expected target genes expressing different microRNAs according to the aging process were input into the gene function analysis of DAVID, KEGG (Kyoto Encyclopedia of a Genes and Genomes) together with the basic parameters limited to the results of the mouse species ( http: / / david.abcc.ncifcrf.gov / ) and Ingenuity Pathway Analysis (Ingenuity Pathway Analysis) ( Systems, http: / / www.ingenuity.com ).

[0134] Table 2

[0135]

[0136]

[0137] As a result, as shown in Table 2 above, a variety of biological effects were derived, mainly expected to be quite significant in the presence of cancer-related pathways, but for the relatively little-studied axon guidance pathway in aging (p = 4.5E- 18) conducted further research. like Figure 2a As shown, it is the same as the schematic diagram of the axon guidance pathway (axon guidance pathway). In the present invention, it is attempted to g...

Embodiment 3

[0138] Example 3. Confirmation of Eph / ephrin pathway regulation according to aging

[0139] 3-1: Confirmation of mRNA expression level

[0140] In the present invention, in order to understand how the expression of Eph / ephrin subfamily (subfamily) changes with aging, real-time fluorescent quantitative polymerase chain reaction analysis was performed.

[0141] First, in order to synthesize cDNA by reverse-transcribing the RNA of the mouse hippocampal tissue isolated in each step in Example 1, PrimeScript was used TM RT reagent Kit (Promega ImPromm-II ReverseTranscription system). Total RNA (1 μg each) was used for generation of cDNA templates according to the manufacturer's instructions, the total reaction volume was 20 μl and the thermal cycler (Bio-Rad Thermal Cycler) from Bio-Rad (Bio-Rad, USA) was used. react.

[0142] Real-time fluorescent quantitative polymerase chain reaction utilizes Bio-Rad real-time PCR system (Bio-Rad real time PCRsystem) (CFX-96) to carry out,...

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Abstract

The present invention relates to a method for judging the functional decline of the hippocampus utilizing the correlation between microRNA (micro RNA; miRNA) and N-methyl-D-aspartate receptor (N-methyl-D-aspartate receptor; NMDAR) . A method for suppressing functional decline and a screening method for functional decline inhibitors. In this specification, miR-204, an upregulated microRNA in the aged hippocampus, correctly targets EphB2, which is an important regulatory receptor, Decrease the expression of EphB2, thereby confirming that the neuron surface expression of the NR1 subunit of the N-methyl-D-aspartate receptor of the hippocampal nerve is reduced, and the density of dendrites is reduced. The relationship between -methyl-D-aspartate receptors determines whether the function of the hippocampus has declined, and further provides the effect of the method for inhibiting the decline in function of the hippocampus and the screening method for inhibitors of function decline.

Description

technical field [0001] This application claims the priority of Korean Patent Application No. 10-2015-0086823 filed on June 18, 2015, and the full text of the above specification is a reference of this application. The present invention relates to the use of microRNA (micro RNA; miRNA) and N-methyl-D-aspartate receptor (N-methyl-D-aspartate receptor; NMDAR) receptor related relationship of hippocampal functional decline Judgment method, method of suppressing functional decline, and screening method of functional decline inhibitor, in more detail, in aging hippocampus (Hippocampus), eph / ephrin mediates axon guidance signal with increased expression of miRNA-204 The pathway (Eph / ephrin mediated axon guidance signaling pathway) is down-regulated, thereby reducing the expression of N-methyl-D-aspartate receptors in the hippocampus, so use their correlation to determine whether the function of the hippocampus is declining, Further provided are a method for suppressing hippocampal f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883A61K45/00A61P25/00
CPCA61K45/00C12Q1/6883C12Q2600/178C12Q2600/136C12Q2600/158C12N15/113G01N33/5058C12N2310/113G01N33/5023A61P25/00C12N2310/11C12N2310/141G01N33/6893
Inventor 金起台南洪吉鼓月
Owner DAEGU GYEONGBUK INST OF SCI & TECH
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