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MiRNA30 cluster as the application of the diagnostic symbol of Alzheimer's disease

An ADAM10, drug technology, applied in the application field of Alzheimer's disease diagnostic markers, can solve the problem of lack of Alzheimer's disease diagnostic markers and other problems

Active Publication Date: 2022-02-18
MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] For this reason, the embodiment of the present invention provides a kind of miRNA biomarker that is used for the diagnosis and / or treatment of Alzheimer's disease, to solve the problem in the prior art that there is no diagnostic marker for Alzheimer's disease at the gene level question

Method used

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  • MiRNA30 cluster as the application of the diagnostic symbol of Alzheimer's disease
  • MiRNA30 cluster as the application of the diagnostic symbol of Alzheimer's disease
  • MiRNA30 cluster as the application of the diagnostic symbol of Alzheimer's disease

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Embodiment 1

[0038] Example 1. miRNA high-throughput microarray technology detects differentially expressed microRNAs in the pathological process of AD

[0039] 1, 3, 6, and 9-month-old double transgenic mice (experimental group) and wild-type mice (control group) stably transfected with APP / PS1 gene were used. Utilizing the principle of Northern blotting, the high-throughput genomics expression profiling biochip technology of "high density, flexible customization, and micro-sample" is carried out, and the method of Trizol is used to extract RNA from mouse brain tissue and separate and label it. The embodiment of the present invention The microRNA of the miRNA 30 cluster is hsa-miR-30a, the sequence of which is shown in SEQ ID NO.1: gcgacuguaaacauccucgacuggaagcugugaagccacagaugggcuuucagucggauguuugcagcugc. Its default mature body (hsa-miR-30a-5p) sequence is shown in SEQ ID NO.2: gaaggucagcuccuacaaaugu. Among them, mature (hsa-miR-30a-5p) miRNA reverse transcription primer: SEQ ID NO.3: gtc...

Embodiment 2

[0041] The expression of the microRNA of embodiment 2, miRNA 30 clusters in AD model cell

[0042] Monoclonal strains were obtained by cell culture technology, liposome transient transfection, antibiotic pressurized screening, and limiting dilution method. At the same time, Western blot or ELISA was used for related protein detection to construct human neurons stably transfected with human-mouse chimeric APP gene. blastoma cells. At the same time, copper ions are used to induce the treatment of cells, copper ions form chelates with APP and Aβ, aggravate the production and deposition of Aβ, and induce oxidative stress response and apoptosis of nerve cells. Therefore, it is used to simulate the pathological state of AD nerve cells and the research on the mechanism of drug action.

[0043] RT-PCR and qPCR techniques were used for reverse transcription and real-time fluorescent quantitative detection of miR-30a expression changes in the pathological process of AD cell models. Su...

Embodiment 3

[0044] The expression of the microRNA of embodiment 3, miRNA 30 clusters in AD model animal

[0045] Double transgenic mice stably transfected with APP / PS1 gene (experimental group) and SAMP8 natural rapid aging mice (control group) were used. The hippocampus and cortex tissues of 1, 3, 6, and 9-month-old APP / PS1 double transgenic mice and SAMP8 natural rapid aging mice were extracted respectively, and the total mRNA of mouse cortex and hippocampus was extracted by Trizol method, and the concentration was determined by ultraviolet absorption method at the same time. and purity determination, using RT-PCR and qPCR techniques to detect the expression changes of miR-30a in the pathological process of AD.

[0046] Such as figure 2 In, B-C, in the cortex or hippocampus of the two animal models, compared with the same-month-old control mice (WTcontrol / SAMR8 mice), the expression levels of miR-30a were significantly different at 1, 3, 6, and 9 months of age A significant increase....

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Abstract

The invention discloses the application of the miRNA30 cluster as a diagnostic marker for Alzheimer's disease. The microRNA of the miRNA 30 cluster of the present invention has a role in the diagnosis and treatment of Alzheimer's disease, and the primers for the microRNA markers of the miRNA30 cluster are used through AD model cells, AD model animals and natural aging animals, and clinical blood samples and / or probes to detect its relative expression in the human body, the expression of the microRNA of the miRNA30 cluster of the present invention is significantly increased in the process of Alzheimer's disease, and negatively regulates the expression of ADAM10 and SIRT1 at the level of transcription and translation , while inhibiting the non-amyloid production pathway mediated by ADAM10 and the amyloid degradation pathway mediated by SIRT1 deacetylation, leading to excessive accumulation of amyloid in the brain and promoting the occurrence and development of AD. Therefore, the microRNA of the miRNA 30 cluster can be used as a new Alzheimer's disease marker for the auxiliary diagnosis of Alzheimer's disease.

Description

technical field [0001] The embodiment of the present invention relates to the field of biotechnology, in particular to the application of miRNA30 cluster as a diagnostic marker for Alzheimer's disease. Background technique [0002] Currently, Alzheimer's disease (AD) is a chronic neurodegenerative disease in the cortex and hippocampus. The pathological mechanism is senile plaque deposition caused by β-amyloid (β-Amyloid-beta peptide, Aβ) aggregation, neurofibrillary tangles caused by hyperphosphorylation of Tau protein, synaptic dysfunction, nerve cell inflammatory response, and cerebral cortical atrophy Wait. There are still problems and challenges in the diagnosis and / or treatment of AD due to the lack of efficient and accurate screening and detection technology, unclear pathogenic mechanism, and slow progress in drug target research. At present, the drugs under clinical research and marketed drugs can only alleviate the symptoms of mild to moderate patients, but cannot ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/11A61K45/00A61P25/28
CPCC12Q1/6883A61K45/00A61P25/28C12Q2600/106C12Q2600/178
Inventor 刘睿李卓荣曾利孙婷姜海伦赵凯悦张俊霞
Owner MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI