CN114748510A - Application of human umbilical cord blood plasma exosome in preparation of medicine for treating degenerative diseases caused by aging - Google Patents
Application of human umbilical cord blood plasma exosome in preparation of medicine for treating degenerative diseases caused by aging Download PDFInfo
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Abstract
The invention provides an application of human umbilical cord blood plasma exosome in preparing a medicament for treating degenerative diseases caused by aging. The kit for treating the degenerative disease caused by aging comprises a human umbilical cord blood plasma exosome. The combination, the method and the kit provided by the invention can be used for treating the degenerative diseases caused by aging, have the advantages of obvious effect, low treatment cost, easy storage of the kit and the like, and can be widely used for relevant work such as treatment of the degenerative diseases caused by aging and become an effective means for treating the degenerative diseases caused by aging.
Description
Technical Field
The invention belongs to the technical field of biology, and relates to application of human umbilical cord blood plasma exosome in preparation of a medicine for treating degenerative diseases caused by aging.
Background
With age, aging occurs in many organs, such as the brain, muscles, bones, etc., with degenerative changes in the nervous system being most pronounced. As the degeneration of organs is gradually increased, a series of functional disorders of the nervous system, the motor system and the like are finally caused, and the diseases are called degenerative diseases. Currently, degenerative diseases mainly include Alzheimer's Disease (AD), Multiple Sclerosis (MS), Parkinson's Disease (PD), Amyotrophic Lateral Sclerosis (ALS), Huntington's Disease (HD), and Spinocerebellar ataxia of different types (SCA).
Blood, as one of the components of the circulatory system, exerts various important physiological functions throughout the body, such as metabolic function, immune function, messenger function, thermoregulation, coagulation function, etc. Researchers find that a dynamic information network (such as various secreted proteins) secreted by tissues and cells exists in human blood through means of proteomics and the like and participates in the aging process. With animal experimentation, some researchers have attempted to search for key factors in the blood that trigger senescence. Such as Conboy, based on the non-long term xenobiotic symbiotic system and real-time blood exchange system of mice, it was demonstrated that some factors in the blood of aging mice can cause aging of some cells and tissues of young mice, while some factors in the blood of young mice have therapeutic effects on aging of some cell tissues of old mice.
Cord blood is the residual blood in the umbilical cord between the mother and the fetus, which contains many active ingredients. The umbilical cord blood can be clinically stored to treat various diseases, such as malignant tumor, blood system diseases, immune diseases and the like, and the umbilical cord blood is an important resource. The abundant hematopoietic stem cells contained in the cord blood are a precious source of hematopoietic stem cells. The hematopoietic function of umbilical cord blood is increasingly recognized by the medical community. The umbilical cord blood transplantation can be used for treating leukemia, diabetes, radiation disease and other diseases which are difficult to cure.
Exosomes (exosomes) are a group of vesicular substances secreted by cells with a diameter of about 40-150nm and a density ranging from 1.09-1.18g/ml, with a double-layered phospholipid membrane structure, containing proteins, mRNA, miRNA and other signal molecules. Many studies on exosomes associated with aging have been reported, but mainly stay at the cellular level. Stimuli such as heat shock, hypoxia, hypothermia, oxidative stress and radiation cause cell aging, thereby increasing secretion of exosomes and promoting changes in bioactive components in exosomes. Research on exosomes associated with aging has focused mainly on the following two aspects: (1) the exosomes secreted by senescent cells are significantly increased in number compared to younger cells. (2) Cellular senescence can significantly alter the composition of exosomes. Exosomes have also been reported to be involved in modulating the development of neurodegenerative diseases. For example, Winston et al examined the role of exosomes of neuronal origin in predicting mild cognitive impairment and AD, and found that the levels of P-tau, Abeta 1-42, neurogenin (neuregulin) and transcription repressor-1 (transcription factor) in these exosomes are significantly altered in the mild cognitive impairment and AD populations compared to the normal population.
At present, the retrogressive diseases caused by aging are mostly targeted at specific targets, have singleness and drug dependence, and cannot relieve the mitochondrial function decline caused by aging from the root.
Disclosure of Invention
In view of the above deficiencies or needs in the art, the present invention provides the use of human umbilical cord blood plasma exosomes for the preparation of a medicament for treating age-induced degenerative diseases.
The purpose of the invention can be realized by the following technical scheme:
application of human umbilical cord blood plasma exosome in preparing medicament for treating degenerative diseases caused by aging.
The application of the human umbilical cord blood plasma exosome in combination with a young human plasma exosome in preparing a medicament for treating degenerative diseases caused by aging.
Preferably, the young people are 18 to 40 years old.
As a further preferred embodiment of the present invention, said exosome is prepared by plasma extraction and/or ultracentrifugation.
A medicine for treating degenerative diseases caused by aging comprises human umbilical cord blood plasma exosome.
As a further preferred embodiment of the present invention, the medicament further comprises young human plasma exosomes.
Has the advantages that:
the technical method for treating mitochondrial dysfunction caused by aging by means of the human umbilical cord blood plasma exosome solves the problems of high production cost, single target spot and the like of the existing neurodegenerative disease medicaments to a great extent, is a low-cost and high-efficiency neurodegenerative disease treatment mode, and simultaneously proves the safety of the exosome treatment mode based on the invention.
Drawings
FIG. 1 shows that the aging index P21 is expressed differently after human umbilical cord blood plasma exosome is co-cultured with NE-4C cells according to gradient.
FIG. 2 shows the differential expression of the aging index P21 in human cord blood plasma exosome and young human plasma exosome co-cultured with NE-4C cells.
FIG. 3 gradient of human umbilical cord blood plasma exosomes with C2C12The condition that the aging index P21 is differentially expressed after cell co-culture.
FIG. 4 human umbilical cord blood plasma exosomes and young human plasma exosomes and C2C12The condition that the aging index P21 is differentially expressed after cell co-culture.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and do not limit the invention. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
The invention provides a plasma exosome extraction kit and application of a plasma exosome injection program.
The method of the plasma exosome extraction kit comprises the following steps:
(1) ultrasonically cleaning a special centrifugal tube of an ultracentrifuge one day in advance, and airing.
(2) The water bath was set to 37 ℃ half an hour prior to operation.
(3) And taking out the subpackaged frozen plasma, thawing in a water bath at 25-37 ℃, and placing on ice after thawing.
(4) Centrifuging at 2-8 deg.C for 2-10 min at 400-700 g, and collecting supernatant.
(5) Centrifuging at 2000-5000 g for 20-30 min at 2-8 deg.C, and collecting supernatant.
(6) Centrifuging at 8000g-13000g for 50-90 min at 2-8 deg.C, and collecting supernatant.
(7) An appropriate amount of PBS was added to the centrifuge tube to dilute the tube as far as possible without air bubbles.
(8) Pouring the plasma into a centrifuge tube, rinsing the tube wall and the tube cover without bubbles, marking outwards, and freely sliding in.
(9) Centrifuging at 100000-160000 g for 60-90 min at 2-8 deg.C with ultracentrifuge.
(10) After finishing, pressing VACUUM, and then opening the door.
(11) The supernatant was aspirated. Note that: the exosomes are in the pellet.
(12) The exosomes were resuspended in the appropriate amount of PBS, and the labeled outer side was carefully rinsed with PBS.
The plasma exosome injection procedure described above comprises the following steps:
(1) collecting exosomes, accurately determining the volume of the exosomes, averagely dividing the exosomes into a plurality of parts, and putting the parts into a freezer for freezing at the temperature of minus 20 ℃;
(2) melting an exosome at room temperature, and injecting the exosome into a living body through a vein;
(3) one injection is injected at intervals, and the injection is performed for at least five times.
Example 2
The influence of the human umbilical cord blood plasma exosome on the aging expression of NE-4C cells was studied by co-culturing the human umbilical cord blood plasma exosome with mouse neural stem cells (NE-4C) in a volume gradient of 25. mu.l, 50. mu.l, 100. mu.l, 200. mu.l and detecting the expression level of the cell aging index P21 by western blot. The results are shown in FIG. 1.
Example 3
The influence of the human umbilical cord blood plasma exosome on the aging expression of NE-4C cells is researched by co-culturing the human umbilical cord blood plasma exosome and the young human plasma exosome with mouse neural stem cells (NE-4C) according to the volume of 25 mul and 50 mul, and detecting the expression level of a cell aging index P21 through western blot. The results are shown in FIG. 2.
Example 4
The influence of human cord blood plasma exosomes on the aging expression of C2C12 cells was studied by co-culturing human cord blood plasma exosomes with mouse myoblasts (C2C12) in a volume gradient of 25. mu.l, 50. mu.l, 100. mu.l, 200. mu.l, and detecting the expression level of the cell aging index P21 by western blot. The results are shown in FIG. 3.
Example 5
The influence of human cord blood plasma exosomes on the aging expression of C2C12 cells was studied by co-culturing human cord blood plasma exosomes and young human plasma exosomes with mouse myoblasts (C2C12) in a volume of 25. mu.l and 50. mu.l, and detecting the expression level of a cell aging index P21 by western blot. The results are shown in FIG. 4.
Claims (6)
1. Application of human umbilical cord blood plasma exosome in preparing medicament for treating degenerative diseases caused by aging.
2. The application of the human umbilical cord blood plasma exosome in combination with a young human plasma exosome in preparing a medicament for treating degenerative diseases caused by aging.
3. The use according to claim 2, wherein the young is between 18 and 40 years of age.
4. Use according to claim 1 or 2, characterized in that said exosomes are prepared by plasma extraction and/or ultracentrifugation.
5. A medicament for treating degenerative diseases caused by aging, which is characterized by comprising human umbilical cord blood plasma exosomes.
6. The medicament of claim 5, wherein said medicament further comprises young human plasma exosomes.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116790492A (en) * | 2023-06-27 | 2023-09-22 | 广州市天河诺亚生物工程有限公司 | Method for producing umbilical cord blood exosomes |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116790492A (en) * | 2023-06-27 | 2023-09-22 | 广州市天河诺亚生物工程有限公司 | Method for producing umbilical cord blood exosomes |
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