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A kind of Parkinson's related biomarker and its application

A Parkinson's disease and drug technology, applied in the field of Parkinson's related biomarkers and biomarkers, can solve the problem that neuroprotective agents cannot effectively improve PD symptoms, etc.

Active Publication Date: 2021-10-01
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A single neuroprotective agent can no longer effectively improve the symptoms of PD. Therefore, according to different risk factors and pathogenic mechanisms, individualized analysis is carried out, so as to formulate targeted regulatory strategies and drug combination treatment plans to delay the progression of PD.

Method used

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  • A kind of Parkinson's related biomarker and its application
  • A kind of Parkinson's related biomarker and its application
  • A kind of Parkinson's related biomarker and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Screening for gene markers associated with Parkinson's disease

[0047] 1. Sample collection

[0048] Collect 10 normal blood samples and blood samples from patients with Parkinson's disease, and write down the sample name, serial number, sampling date, sample processing process, etc. All the above-mentioned samples were obtained with the approval of the ethics committee.

[0049] 2. Preparation of RNA samples

[0050] RNA samples were extracted using Invitrogen's Blood RNA Extraction Kit. For details, see the instruction manual.

[0051] 3. Quality analysis of RNA samples

[0052] The above-mentioned extracted RNA was subjected to agarose gel electrophoresis, and the concentration and purity of the extracted RNA were detected by Nanodrop2000, the RNA integrity was detected by agarose gel electrophoresis, and the RIN value was determined by Agilent2100. The total amount of RNA required for a single library construction is 5ug, the concentration is ≥200ng / μL...

Embodiment 2

[0063] Example 2 QPCR sequencing to verify the differential expression of the FGFBP2 gene

[0064] 1. According to the detection results of high-throughput sequencing, the FGFBP2 gene was selected for large-sample QPCR verification. According to the sample collection method in Example 1, 90 cases of blood from Parkinson's patients and 90 cases of normal blood were selected.

[0065] 2. The RNA extraction steps are the same as in Example 1.

[0066] 3. Reverse transcription:

[0067] (1) Take 2 μg of total RNA for reverse transcription, add 2 μl of Oligo(dT), and mix well. After 5 minutes in water bath at 70°C, immediately ice bath for 2-3 minutes.

[0068] (2) Construct a 25 μl reaction system, including 5 μl of 5× reverse transcription buffer, 5 μl of dNTP (2.5 mM), 40 U / μl of RNasin, 200 U / μl of M-MLV, and make up to the expected volume with nuclease-free water.

[0069] (3) After 60 minutes of water bathing at 42° C., then 5 minutes of water bathing at 95° C. to inactiv...

Embodiment 3

[0090] Example 3 Silencing of FGFBP2 Gene

[0091] 1. Cell culture

[0092] Dopamine neuronal cells SH-SY5Y were prepared in DMEM medium containing 10% fetal bovine serum and 1% penicillin / streptomycin (pH 7.2-7.4), at 37°C and 5% CO 2 , Cultivated in an incubator with a relative humidity of 90%. Change the medium once every 2 days, and subculture when the cells grow to 90% contact. After washing with PBS, add 0.25%-EDTA trypsin to separate the cells from the bottle wall, and terminate with DMEM medium containing fetal bovine serum. Trypsin digestion reaction, centrifuged at 1000g for 2min, discarded the supernatant, resuspended with the newly prepared culture medium, and passaged at a ratio of 1:3 to 1:4. After 24 hours, the cells entered the logarithmic growth phase and replaced the culture medium according to the experimental requirements. give different interventions.

[0093] 2. siRNA design

[0094] siRNA sequences against FGFBP2:

[0095] siRNA1:

[0096] The sens...

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Abstract

The invention discloses a Parkinson-related biomarker and its application. It is proved by experiments that FGFBP2 is up-regulated in the blood of Parkinson's patients, suggesting that FGFBP2 can be used as a detection index in the clinical diagnosis of Parkinson's; The expression level of FGFBP2 can affect the proliferation of nerve cells, suggesting that FGFBP2 can be used as a drug target in the personalized treatment of Parkinson's patients.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to a Parkinson-related biomarker and application thereof, specifically, the biomarker is FGFBP2. Background technique [0002] Neurodegenerative diseases seriously endanger the health of the elderly. The World Health Organization announced that with the increasingly prominent phenomenon of population aging, neurodegenerative diseases will become the second largest disease that endangers human health after cardiovascular diseases by 2040 ( . Valera E, Masliah E. Therapeutic approaches in Parkinson's disease and related disorders. Journal of neurochemistry, (2016)). Parkinson's disease (PD) is the second most common neurodegenerative disease. It is urgent to further understand the pathological mechanism of PD and develop effective treatments. The main pathological feature of PD is the progressive loss of dopamine (DA) neurons in the substantia nigra compacta of the midbrain. Clinicopatholo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/68A61K31/713A61P25/16
CPCA61K31/713A61P25/16C12Q1/6886C12Q2600/178G01N33/68G01N2800/2835
Inventor 肖枫汪冰怡
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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