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Application of miRNA-543-3p in diagnosis and treatment of Parkinson's disease

A Parkinson's disease and inhibitor technology, applied in the application field of miRNA-543-3p in the diagnosis and treatment of Parkinson's disease, to achieve the effect of reducing neurotoxic effects, relieving the disease, and weakening the inhibitory effect

Inactive Publication Date: 2016-06-01
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the microRNAs that regulate glutamate transporters in Parkinson's disease have not been reported so far, and it is worthy of our further exploration

Method used

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  • Application of miRNA-543-3p in diagnosis and treatment of Parkinson's disease
  • Application of miRNA-543-3p in diagnosis and treatment of Parkinson's disease
  • Application of miRNA-543-3p in diagnosis and treatment of Parkinson's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Preparation and pathological research of embodiment 1PD mouse model

[0049] 1. Preparation and identification of PD mouse model

[0050] Preparation of PD mouse model by MPTP:

[0051] 12-week-old healthy male C57 / bl mice (body weight about 25-30g) were randomly divided into normal control group and MPTP-built PD model group. Intraperitoneal injection of MPTP20mg / kg, once a day, a total of 5 days. On the 3rd day after the injection and the 3rd day and the 5th day after the injection, the modeling was tested by animal behavior and immunohistochemical staining.

[0052] The detection content includes:

[0053] (1) Rod climbing test to detect the behavioral changes of mice.

[0054] A foam plastic ball with a diameter of 2.5 cm is fixed on the top of a wooden pole with a length of 50 cm and a thickness of 1 cm, and the wooden pole is wrapped with 2 layers of gauze to prevent slipping. Put the mouse on the top of the ball, and record the following three times: the tim...

Embodiment 2

[0072] Example 2 Screening of differentially expressed microRNAs and comparison with bioinformatics prediction results

[0073] 1. Screening and verification of differentially expressed microRNAs by high-throughput sequencing

[0074] (1) High-throughput sequencing to screen differentially expressed microRNAs

[0075] method:

[0076] Three days after the completion of modeling, the mice were sacrificed, and the brain and midbrain were separated to extract RNA from each group. The differentially expressed microRNAs of mice in each group were completed by high-throughput sequencing technology. The principle of this technology is to perform end repair on the fragments of microRNAs in the sample to be tested and connect the 5'-adapter and 3'-adapter at both ends, and perform computer detection and data processing to screen out microRNAs with significant expression differences.

[0077] 1) Selection of high-throughput sequencing: The high-throughput sequencing technology select...

Embodiment 3

[0128] Functional analysis of embodiment 3mmu-miR-543-3p

[0129] 1. Cell experiments to detect the regulatory effect of mmu-miR-543-3p on GLT-1

[0130] Overexpression of mmu-miR-543-3pmimics (SEQ ID NO: 2) and specific inhibition of mmu-miR-543-3 pinhibitors (SEQ ID NO: 3) by micro RNA analysis of microRNA for MPP + Regulation of GLT-1 expression and function on infected astrocytes. Wherein, mmu-miR-543-3pinhibitors is the antisense oligonucleotide sequence of mmu-miR-543-3p, that is, the inhibitor.

[0131] method:

[0132] 1) Culture of mouse primary astrocytes: newborn C57 / bl mice born 1 to 3 days after birth were taken out, and astrocytes were prepared and further purified and identified.

[0133] 2) Transfect normal mouse astrocytes and MPP with mmu-miR-543-3pinhibitors and mmu-miR-543-3pmimics respectively + The mouse astrocytes in the exposure group were cultured for 48 hours to detect the following indicators:

[0134] ①Using qPCR to detect the mRNA expression o...

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Abstract

The invention discloses an application of miRNA-543-3p in diagnosis and treatment of a Parkinson's disease. In the invention, it is screened and verified that the miRNA-543-3p has an effect of regulating expressions and functions of a glutamate transporter, and it is further verified that miRNA-543-5p directly acts on the glutamate transporter. It is discovered that miRNA-543-5p inhibitors can inhibit functions of the miRNA-543-5p, so that an inhibition effect of the miRNA-543-5p on mRNA translation of the glutamate transporter is reduced, and an expression level of a glutamate transporter protein is improved to some extent, thereby reducing a neurotoxicity effect of glutamic acid, relieving the state of the Parkinson's disease and providing a new target and a new way for clinical treatment of the Parkinson's disease.

Description

technical field [0001] The invention relates to the application of miRNA-543-3p in the diagnosis and treatment of Parkinson's disease. Background technique [0002] Parkinson's disease (PD) is a common neurodegenerative disease. Patients will experience resting tremor, muscle rigidity, slow movement, and abnormal posture and gait. Many patients also have symptoms such as anosmia, depression, and dementia. The condition is progressive It is one of the common disabling diseases of middle-aged and elderly people, which seriously affects the quality of life of patients and brings heavy burdens to society and families. The main pathological changes of Parkinson's disease are progressive degeneration and death of dopaminergic neurons in the substantia nigra of the midbrain; eosinophilic protein inclusion bodies (Lewybody, LB) appear in the remaining neurons, and dopamine in the striatum significantly decreases. Although the etiology of PD is still unclear, studies on various anim...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K48/00A61K31/7105A61P25/16C12Q1/68
CPCA61K45/00A61K31/7105C12Q1/6883C12Q2600/158
Inventor 瞿少刚张云龙何小亮张秀萍吴晓娟童辉纯陆伶俐秦笙
Owner SOUTHERN MEDICAL UNIVERSITY
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