Application of young blood plasma exosome in preparation of medicine for treating senescence-induced degenerative diseases

A technology for degenerative diseases, young people, applied in the biological field, can solve problems such as mitochondrial function decline

Pending Publication Date: 2022-07-15
NANJING YOUZHIYUAN PHARMACEUTICAL TECHNOLOGY CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, degenerative diseases caused by aging are mostly aimed at specific targets, which are single and drug-dependent, and cannot fundamentally alleviate the decline of mitochondrial function caused by aging

Method used

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  • Application of young blood plasma exosome in preparation of medicine for treating senescence-induced degenerative diseases
  • Application of young blood plasma exosome in preparation of medicine for treating senescence-induced degenerative diseases
  • Application of young blood plasma exosome in preparation of medicine for treating senescence-induced degenerative diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Extraction of plasma exosomes

[0019] The method of the above-mentioned plasma exosome extraction kit comprises the following steps:

[0020] (1) Ultrasonic cleaning of the ultracentrifuge special centrifuge tube one day in advance, and drying.

[0021] (2) Set the water bath to 37°C half an hour before the operation.

[0022] (3) Take out the subpackaged frozen plasma, thaw in a water bath at 25-37°C, and place it on ice after thawing.

[0023] (4) Centrifuge at 500g for 5 minutes at 4°C, and take the supernatant.

[0024] (5) Centrifuge at 3000g for 25 minutes at 4°C, and take the supernatant.

[0025] (6) Centrifuge at 10,000g for 60 minutes at 4°C, and take the supernatant.

[0026] (7) First add an appropriate amount of PBS to the 15ml centrifuge tube to dilute, try not to have air bubbles.

[0027] (8) Pour the plasma into the centrifuge tube without air bubbles. Rinse the tube wall and tube cover without air bubbles. The mark faces outward and sli...

Embodiment 2

[0037] After extracting the plasma exosomes from young mice, the exosomes were injected into the old C57BL / 6J mice according to the plasma exosome injection procedure shown in Example 1, and then the mouse hippocampus and muscle tissue proteins were extracted and detected by western bolt Changes of differentially expressed genes in each tissue, see the results figure 1 .

Embodiment 3

[0039] After the plasma exosomes were extracted, the exosomes were co-cultured with nerve cells (NE4C) and muscle cells (C2C12), respectively, and the following indicators were detected after 24 hours:

[0040] (1) The changes of differentially expressed genes in cells were detected by western bolt;

[0041] (2) Detection of cell proliferation ability by EdU;

[0042] (3) The mRNA expression of maker P21, a marker of aging, was detected by qPCR;

[0043] (4) Detect the change of ATP generation rate by ATP generation rate detection kit;

[0044] (5) Detect the changes of mitochondrial complex V activity by the mitochondrial complex V activity detection kit;

[0045] (6) Detection of mitochondrial DNA changes by qPCR;

[0046] (7) The changes of mitochondrial respiratory chain were detected by seahorse instrument mitochondrial stress kit.

[0047] see the results figure 2 .

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Abstract

The invention relates to an application of a young plasma exosome in preparation of a medicine for treating degenerative diseases caused by senescence. The exosome extracted from blood plasma of young people is used for treating degenerative diseases caused by aging, and has the advantages of obvious effect, low treatment cost, easiness in storage of the kit and the like; the technical method for treating mitochondrial dysfunction caused by aging by relying on young plasma exosomes solves the problems of high production cost, single target spot and the like of the existing neurodegenerative disease medicines to a great extent, and is a low-cost and high-efficiency neurodegenerative disease treatment mode; meanwhile, the safety of an exosome treatment mode based on the invention is proved. The method can be widely applied to related work such as treatment of senescence-induced degenerative diseases, and becomes an effective means for treatment of senescence-induced degenerative diseases.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the application of young plasma exosomes in the preparation of medicines for treating degenerative diseases caused by aging. Background technique [0002] With the increase of age, aging will occur in many organs, such as the brain, muscles, bones, etc., among which the degenerative changes of the nervous system are the most obvious. As the degenerative changes of organs gradually intensify, it will eventually lead to a series of functional disorders such as the nervous system and motor system, which are called degenerative diseases. At present, degenerative diseases mainly include Alzheimer's disease (AD), Multiple sclerosis (MS), Parkinson's Disease (PD), Amyotrophic lateral sclerosis (Amyotrophic lateral sclerosis, ALS), Huntington disease (HD), and different types of Spinocerebellar Atrophy (SCA). [0003] As one of the components of the circulatory system, blood plays a variety ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/16A61P39/06A61P25/28
CPCA61K35/16A61P39/06A61P25/28
Inventor 朱卿骆阳陈骁锐
Owner NANJING YOUZHIYUAN PHARMACEUTICAL TECHNOLOGY CO LTD
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